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PMC:3317373 / 1291-20739 JSONTXT 26 Projects

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Id Subject Object Predicate Lexical cue
T13 0-2 Sentence denotes 1.
T14 3-15 Sentence denotes Introduction
T620 16-149 Sentence denotes Brain damage following traumatic injury is a result of direct (primary injury) and indirect (secondary or delayed injury) mechanisms.
T15 16-149 Sentence denotes Brain damage following traumatic injury is a result of direct (primary injury) and indirect (secondary or delayed injury) mechanisms.
T621 150-494 Sentence denotes The secondary injury mechanism involves the initiation of an acute inflammatory response, including breakdown of blood-brain barrier (BBB), brain edema, infiltration of peripheral blood cells, and activation of resident immunocompetent cells, as well as the release of numerous immune mediators such as interleukins and chemotactic factors [1].
T16 150-494 Sentence denotes The secondary injury mechanism involves the initiation of an acute inflammatory response, including breakdown of blood-brain barrier (BBB), brain edema, infiltration of peripheral blood cells, and activation of resident immunocompetent cells, as well as the release of numerous immune mediators such as interleukins and chemotactic factors [1].
T622 495-602 Sentence denotes And the subsequent inflammation leads to secondary damage not only in brain but also in other organs [2–4].
T17 495-602 Sentence denotes And the subsequent inflammation leads to secondary damage not only in brain but also in other organs [2–4].
T623 603-726 Sentence denotes Nuclear factor erythroid 2-related factor2 (Nrf2) is a transcription factor that regulates many kinds of antioxidant genes.
T18 603-726 Sentence denotes Nuclear factor erythroid 2-related factor2 (Nrf2) is a transcription factor that regulates many kinds of antioxidant genes.
T624 727-846 Sentence denotes Several studies have demonstrated that Nrf2 regulates the inflammation in the brain after traumatic brain injury (TBI).
T19 727-846 Sentence denotes Several studies have demonstrated that Nrf2 regulates the inflammation in the brain after traumatic brain injury (TBI).
T625 847-959 Sentence denotes It was also showed that TBI could induce more aggravated damage in Nrf2 knockout mice than in wildtype mice [5].
T20 847-959 Sentence denotes It was also showed that TBI could induce more aggravated damage in Nrf2 knockout mice than in wildtype mice [5].
T626 960-1097 Sentence denotes On the contrary, such inflammation can be extenuated through elevating the level of Nrf2 by its inducers, such as sulforaphane (SFN) [6].
T21 960-1097 Sentence denotes On the contrary, such inflammation can be extenuated through elevating the level of Nrf2 by its inducers, such as sulforaphane (SFN) [6].
T627 1098-1232 Sentence denotes It is well documented that astrocytes play a critical role in maintaining normal brain physiology and responding to injury or disease.
T22 1098-1232 Sentence denotes It is well documented that astrocytes play a critical role in maintaining normal brain physiology and responding to injury or disease.
T628 1233-1295 Sentence denotes In fact, all the aforementioned studies are performed in vivo.
T23 1233-1295 Sentence denotes In fact, all the aforementioned studies are performed in vivo.
T629 1296-1389 Sentence denotes Little is known about the relationship between Nrf2 and inflammation in astrocytes after TBI.
T24 1296-1389 Sentence denotes Little is known about the relationship between Nrf2 and inflammation in astrocytes after TBI.
T630 1390-1560 Sentence denotes Therefore, here we analysed the influence of Nrf2 on the expression of proinflammatory cytokines in primary cultured astrocytes from transgenic mice after scratch injury.
T25 1390-1560 Sentence denotes Therefore, here we analysed the influence of Nrf2 on the expression of proinflammatory cytokines in primary cultured astrocytes from transgenic mice after scratch injury.
T26 1562-1564 Sentence denotes 2.
T27 1565-1586 Sentence denotes Methods and Materials
T1224 1588-1592 Sentence denotes 2.1.
T28 1588-1592 Sentence denotes 2.1.
T1225 1593-1647 Sentence denotes Primary Culture and Identification of Mouse Astrocytes
T29 1593-1647 Sentence denotes Primary Culture and Identification of Mouse Astrocytes
T30 1648-1708 Sentence denotes Nrf2 knockout ICR mice were kindly provided by Dr. Thomas W.
T1226 1648-1764 Sentence denotes Nrf2 knockout ICR mice were kindly provided by Dr. Thomas W. Kensler (Johns Hopkins University, Baltimore, MD, USA).
T31 1709-1764 Sentence denotes Kensler (Johns Hopkins University, Baltimore, MD, USA).
T1227 1765-1893 Sentence denotes Primary astrocytes were obtained from postnatal 2-day-old Nrf2 wide type (WT) and knockout (KO) mice (6 mice for each genotype).
T32 1765-1893 Sentence denotes Primary astrocytes were obtained from postnatal 2-day-old Nrf2 wide type (WT) and knockout (KO) mice (6 mice for each genotype).
T1228 1894-2010 Sentence denotes Following decapitation, the cortices were dissected out, and the meninges and associated blood vessels were removed.
T33 1894-2010 Sentence denotes Following decapitation, the cortices were dissected out, and the meninges and associated blood vessels were removed.
T1229 2011-2145 Sentence denotes The tissue was roughly chopped with a scalpel blade, incubated in 0.5% trypsin for 10 minutes at 37°C, and agitated every few minutes.
T34 2011-2145 Sentence denotes The tissue was roughly chopped with a scalpel blade, incubated in 0.5% trypsin for 10 minutes at 37°C, and agitated every few minutes.
T1230 2146-2346 Sentence denotes After digestion, the tissue was rinsed twice in DMEM with 10% FBS, followed by a mechanical dissociation in DMEM with 20% FBS and 5 units/mL penicillin, 5 μg/mL streptomycin (complete culture medium).
T35 2146-2346 Sentence denotes After digestion, the tissue was rinsed twice in DMEM with 10% FBS, followed by a mechanical dissociation in DMEM with 20% FBS and 5 units/mL penicillin, 5 μg/mL streptomycin (complete culture medium).
T1231 2347-2486 Sentence denotes After incubation for 1 h, the supernatant was transferred to a new flask or dish (Costar, USA) for depleting the residual epithelial cells.
T36 2347-2486 Sentence denotes After incubation for 1 h, the supernatant was transferred to a new flask or dish (Costar, USA) for depleting the residual epithelial cells.
T1232 2487-2538 Sentence denotes Then the cells were cultivated at 37°C with 5% CO2.
T37 2487-2538 Sentence denotes Then the cells were cultivated at 37°C with 5% CO2.
T1233 2539-2601 Sentence denotes And the complete culture medium was half-changed twice a week.
T38 2539-2601 Sentence denotes And the complete culture medium was half-changed twice a week.
T1234 2602-2659 Sentence denotes Astrocytes expanded for about 7 days to reach confluence.
T39 2602-2659 Sentence denotes Astrocytes expanded for about 7 days to reach confluence.
T1235 2660-2784 Sentence denotes Then the flasks or dishes were shaken at 150 rpm for 4 h to deplete the microglia and less adherent cells from the cultures.
T40 2660-2784 Sentence denotes Then the flasks or dishes were shaken at 150 rpm for 4 h to deplete the microglia and less adherent cells from the cultures.
T1236 2785-2945 Sentence denotes After shaking, the resulting cultures were mainly astrocytes, which were determined by immunoreactivity for GFAP (sc-166481, Santa Cruz Biotechnology, CA, USA).
T41 2785-2945 Sentence denotes After shaking, the resulting cultures were mainly astrocytes, which were determined by immunoreactivity for GFAP (sc-166481, Santa Cruz Biotechnology, CA, USA).
T1237 2946-3016 Sentence denotes Cells passaged for 2-3 generations were used in the following studies.
T42 2946-3016 Sentence denotes Cells passaged for 2-3 generations were used in the following studies.
T1813 3018-3022 Sentence denotes 2.2.
T43 3018-3022 Sentence denotes 2.2.
T1814 3023-3074 Sentence denotes In Vitro Model of TBI Established by Scratch Injury
T44 3023-3074 Sentence denotes In Vitro Model of TBI Established by Scratch Injury
T1815 3075-3142 Sentence denotes Astrocyte scratch injury was performed as in a previous report [7].
T45 3075-3142 Sentence denotes Astrocyte scratch injury was performed as in a previous report [7].
T1816 3143-3208 Sentence denotes Astrocytes were planted in 6-well plates and grown to confluence.
T46 3143-3208 Sentence denotes Astrocytes were planted in 6-well plates and grown to confluence.
T1817 3209-3325 Sentence denotes The cell monolayer was scratched with a sterile 26G syringe needle, resulting in the formation of a 0.5 mm wide gap.
T47 3209-3325 Sentence denotes The cell monolayer was scratched with a sterile 26G syringe needle, resulting in the formation of a 0.5 mm wide gap.
T1818 3326-3457 Sentence denotes Immediately after scratch, cells were washed twice with sterile PBS, cultured with complete culture medium, and named as TBI group.
T48 3326-3457 Sentence denotes Immediately after scratch, cells were washed twice with sterile PBS, cultured with complete culture medium, and named as TBI group.
T1819 3458-3548 Sentence denotes Cells, which did not received scratch, were used as blank control and named as sham group.
T49 3458-3548 Sentence denotes Cells, which did not received scratch, were used as blank control and named as sham group.
T2033 3550-3554 Sentence denotes 2.3.
T50 3550-3554 Sentence denotes 2.3.
T2034 3555-3592 Sentence denotes Cell Death Measurement by CCK-8 Assay
T51 3555-3592 Sentence denotes Cell Death Measurement by CCK-8 Assay
T2035 3593-3684 Sentence denotes 1 × 103/well cells were seeded to 96-well culture plates and cultivated for 24 h to adhere.
T52 3593-3684 Sentence denotes 1 × 103/well cells were seeded to 96-well culture plates and cultivated for 24 h to adhere.
T2036 3685-3730 Sentence denotes Then the cell was scrated as mentioned above.
T53 3685-3730 Sentence denotes Then the cell was scrated as mentioned above.
T2037 3731-3879 Sentence denotes Astrocyte cell death was assessed by cell counting kit-8 (CCK-8) (Dojindo, Japan) assay 24 h after scratch according to the manufacturer's protocol.
T54 3731-3879 Sentence denotes Astrocyte cell death was assessed by cell counting kit-8 (CCK-8) (Dojindo, Japan) assay 24 h after scratch according to the manufacturer's protocol.
T2038 3880-3949 Sentence denotes Briefly, 10 μL CCK-8 was added into every well and incubated for 1 h.
T55 3880-3949 Sentence denotes Briefly, 10 μL CCK-8 was added into every well and incubated for 1 h.
T2039 3950-4055 Sentence denotes Then OD value was read at 450 nm using a Bio-Rad ELISA microplate reader (Bio-Rad Laboratories, CA, USA).
T56 3950-4055 Sentence denotes Then OD value was read at 450 nm using a Bio-Rad ELISA microplate reader (Bio-Rad Laboratories, CA, USA).
T2040 4056-4104 Sentence denotes All measurements were performed in sextuplicate.
T57 4056-4104 Sentence denotes All measurements were performed in sextuplicate.
T2041 4105-4163 Sentence denotes Results were expressed as mean of OD value at 450 nm ± SD.
T58 4105-4163 Sentence denotes Results were expressed as mean of OD value at 450 nm ± SD.
T2309 4165-4169 Sentence denotes 2.4.
T59 4165-4169 Sentence denotes 2.4.
T2310 4170-4213 Sentence denotes Electrophoretic Mobility Shift Assay (EMSA)
T60 4170-4213 Sentence denotes Electrophoretic Mobility Shift Assay (EMSA)
T2311 4214-4300 Sentence denotes Monolayers of astrocytes were washed with PBS and harvested by scraping into cold PBS.
T61 4214-4300 Sentence denotes Monolayers of astrocytes were washed with PBS and harvested by scraping into cold PBS.
T2312 4301-4493 Sentence denotes The cell pellet obtained by centrifugation was resuspended in buffer containing 10 mM HEPES (pH 7.9), 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1.0 mM DTT, and 0.5 mM phenylmethylsulfonyl fluoride.
T62 4301-4493 Sentence denotes The cell pellet obtained by centrifugation was resuspended in buffer containing 10 mM HEPES (pH 7.9), 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1.0 mM DTT, and 0.5 mM phenylmethylsulfonyl fluoride.
T2313 4494-4595 Sentence denotes Then 10% Nonidet P-40 was added and vortexed briefly, and the nuclei were pelleted by centrifugation.
T63 4494-4595 Sentence denotes Then 10% Nonidet P-40 was added and vortexed briefly, and the nuclei were pelleted by centrifugation.
T2314 4596-4769 Sentence denotes The nuclear proteins were extracted with buffer containing 20 mM HEPES (pH 7.9), 0.4 mM NaCl, 1.0 mM EDTA, 1.0 mM EGTA, 1.0 mM DTT, and 1.0 mM phenylmethylsulfonyl fluoride.
T64 4596-4769 Sentence denotes The nuclear proteins were extracted with buffer containing 20 mM HEPES (pH 7.9), 0.4 mM NaCl, 1.0 mM EDTA, 1.0 mM EGTA, 1.0 mM DTT, and 1.0 mM phenylmethylsulfonyl fluoride.
T2315 4770-4915 Sentence denotes Insoluble material was removed by centrifugation at 14000 rpm, and the supernatant containing the nuclear proteins was stored at –80°C until use.
T65 4770-4915 Sentence denotes Insoluble material was removed by centrifugation at 14000 rpm, and the supernatant containing the nuclear proteins was stored at –80°C until use.
T2316 4916-5073 Sentence denotes Protein concentration was determined using a bicinchoninic acid assay kit with bovine serum albumin as the standard (Pierce Biochemicals, Rockford, IL, USA).
T66 4916-5073 Sentence denotes Protein concentration was determined using a bicinchoninic acid assay kit with bovine serum albumin as the standard (Pierce Biochemicals, Rockford, IL, USA).
T2317 5074-5150 Sentence denotes EMSA was performed using gel shift assay system (Promega, Madison, WI, USA).
T67 5074-5150 Sentence denotes EMSA was performed using gel shift assay system (Promega, Madison, WI, USA).
T2318 5151-5308 Sentence denotes Consensus oligonucleotide probe (5′-AGTTGAGGGGACTTTCCCAGGG-3′) was end-labeled by T4-polynucleotide kinase using [γ-32P]-ATP (Free Biotech., Beijing, China).
T68 5151-5308 Sentence denotes Consensus oligonucleotide probe (5′-AGTTGAGGGGACTTTCCCAGGG-3′) was end-labeled by T4-polynucleotide kinase using [γ-32P]-ATP (Free Biotech., Beijing, China).
T2319 5309-5515 Sentence denotes Nuclear protein (20 μg) was preincubated in 20 μL binding buffer containing 10 mM Tris-HCl (PH 7.5), 1 mM MgCl2, 0.5 mM NaCl, 4% glycerol, 0.5 mM EDTA, 0.5 mM DTT, and 2 μg poly dI-dC for 20 minutes on ice.
T69 5309-5515 Sentence denotes Nuclear protein (20 μg) was preincubated in 20 μL binding buffer containing 10 mM Tris-HCl (PH 7.5), 1 mM MgCl2, 0.5 mM NaCl, 4% glycerol, 0.5 mM EDTA, 0.5 mM DTT, and 2 μg poly dI-dC for 20 minutes on ice.
T2320 5516-5628 Sentence denotes After addition of the 1 μL 32P-labled oligonucleotide probe, the incubation was continued for 20 minutes on ice.
T70 5516-5628 Sentence denotes After addition of the 1 μL 32P-labled oligonucleotide probe, the incubation was continued for 20 minutes on ice.
T71 5629-5794 Sentence denotes The DNA-protein complexes were separated by electrophoresis on 4% nondenaturing polyacrylamide gel in 0.5 × TBE buffer (tris-borate-EDTA) at 390 V for 1 hour at 4°C.
T2321 5629-5893 Sentence denotes The DNA-protein complexes were separated by electrophoresis on 4% nondenaturing polyacrylamide gel in 0.5 × TBE buffer (tris-borate-EDTA) at 390 V for 1 hour at 4°C. After electrophoresis, the gel was dried and exposed to X-ray film (Fuji Hyperfilm, Tokyo, Japan).
T72 5795-5893 Sentence denotes After electrophoresis, the gel was dried and exposed to X-ray film (Fuji Hyperfilm, Tokyo, Japan).
T2322 5894-5966 Sentence denotes Levels of NF-κB DNA binding activity were quantified by software ImageJ.
T73 5894-5966 Sentence denotes Levels of NF-κB DNA binding activity were quantified by software ImageJ.
T3100 5968-5972 Sentence denotes 2.5.
T74 5968-5972 Sentence denotes 2.5.
T3101 5973-5979 Sentence denotes RT-PCR
T75 5973-5979 Sentence denotes RT-PCR
T3102 5980-6192 Sentence denotes Total RNA was isolated with Trizol (Invitrogen, CA, USA), and single-stranded cDNA was synthesized from 2 μg of total RNA with BU-Script RT-Kit (Biunique, Jiangsu, China) according to the manufacturer's protocol.
T76 5980-6192 Sentence denotes Total RNA was isolated with Trizol (Invitrogen, CA, USA), and single-stranded cDNA was synthesized from 2 μg of total RNA with BU-Script RT-Kit (Biunique, Jiangsu, China) according to the manufacturer's protocol.
T77 6193-6222 Sentence denotes The cDNA was stored in −20°C.
T3103 6193-6347 Sentence denotes The cDNA was stored in −20°C. Reverse transcription was conducted with GoTaq Green Master Mix (Promega, WI, USA) according to the manufacturer's protocol.
T78 6223-6347 Sentence denotes Reverse transcription was conducted with GoTaq Green Master Mix (Promega, WI, USA) according to the manufacturer's protocol.
T3104 6349-6394 Sentence denotes Table 1 shows the primers and PCR parameters.
T79 6349-6394 Sentence denotes Table 1 shows the primers and PCR parameters.
T3105 6395-6453 Sentence denotes PCR products were detected by agarose gel electrophoresis.
T80 6395-6453 Sentence denotes PCR products were detected by agarose gel electrophoresis.
T3106 6454-6512 Sentence denotes The intensity of the bands was analyzed by ImageJ program.
T81 6454-6512 Sentence denotes The intensity of the bands was analyzed by ImageJ program.
T3107 6513-6567 Sentence denotes The level of β-actin was used as an internal standard.
T82 6513-6567 Sentence denotes The level of β-actin was used as an internal standard.
T3374 6569-6573 Sentence denotes 2.6.
T83 6569-6573 Sentence denotes 2.6.
T3375 6574-6586 Sentence denotes Western Blot
T84 6574-6586 Sentence denotes Western Blot
T85 6587-6866 Sentence denotes To obtain total protein lysates, cells were homogenized in RIPA buffer (1% NP40, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM EDTA, 1 mM EGTA, 1 mM Na3VO4, 20 mM NaF, 0.5 mM DTT, 1 mM PMSF, and protease inhibitor cocktail in PBS pH 7.4) and centrifuged at 12,000 g for 15 min at 4°C.
T3376 6587-6963 Sentence denotes To obtain total protein lysates, cells were homogenized in RIPA buffer (1% NP40, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM EDTA, 1 mM EGTA, 1 mM Na3VO4, 20 mM NaF, 0.5 mM DTT, 1 mM PMSF, and protease inhibitor cocktail in PBS pH 7.4) and centrifuged at 12,000 g for 15 min at 4°C. Protein concentrations were estimated by Coomassie Plus Protein Assay Reagent (Pierce, IL, USA).
T86 6867-6963 Sentence denotes Protein concentrations were estimated by Coomassie Plus Protein Assay Reagent (Pierce, IL, USA).
T3377 6964-7205 Sentence denotes Fifty micrograms of the resulting cytosolic protein extracts were heat-denatured in Laemmli sample loading buffer, separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and electroblotted onto a nitrocellulose membrane.
T87 6964-7205 Sentence denotes Fifty micrograms of the resulting cytosolic protein extracts were heat-denatured in Laemmli sample loading buffer, separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and electroblotted onto a nitrocellulose membrane.
T3378 7206-7511 Sentence denotes For immunoblotting, membranes were blocked with 5% nonfat dry milk in saline buffer overnight at 4°C, and the following antibodies were used: anti-β-actin (sc-130657, Santa Cruz Biotechnology, CA, USA, 43 kDa) and anti-matrix metallopeptidase 9 (MMP9) (sc-6841, Santa Cruz Biotechnology, CA, USA, 92 kDa).
T88 7206-7511 Sentence denotes For immunoblotting, membranes were blocked with 5% nonfat dry milk in saline buffer overnight at 4°C, and the following antibodies were used: anti-β-actin (sc-130657, Santa Cruz Biotechnology, CA, USA, 43 kDa) and anti-matrix metallopeptidase 9 (MMP9) (sc-6841, Santa Cruz Biotechnology, CA, USA, 92 kDa).
T3379 7512-7635 Sentence denotes Each primary antibody was diluted appropriately in blocking buffer and then added to the blots for 1 h at room temperature.
T89 7512-7635 Sentence denotes Each primary antibody was diluted appropriately in blocking buffer and then added to the blots for 1 h at room temperature.
T3380 7636-7793 Sentence denotes The blots were washed three times in the washing buffer and covered with the horseradish peroxidase-linked secondary antibody at a 1 : 2000 dilution for 1 h.
T90 7636-7793 Sentence denotes The blots were washed three times in the washing buffer and covered with the horseradish peroxidase-linked secondary antibody at a 1 : 2000 dilution for 1 h.
T3381 7794-7968 Sentence denotes Blots were incubated with enhanced chemiluminescence (ECL) detection system (Amersham Biosciences, Bucks, UK) and exposed to radiographic film (Fuji Hyperfilm, Tokyo, Japan).
T91 7794-7968 Sentence denotes Blots were incubated with enhanced chemiluminescence (ECL) detection system (Amersham Biosciences, Bucks, UK) and exposed to radiographic film (Fuji Hyperfilm, Tokyo, Japan).
T3382 7969-8023 Sentence denotes ImageJ was used to analyze the intensity of the blots.
T92 7969-8023 Sentence denotes ImageJ was used to analyze the intensity of the blots.
T3383 8024-8075 Sentence denotes The level of β-actin was used as internal standard.
T93 8024-8075 Sentence denotes The level of β-actin was used as internal standard.
T4052 8077-8081 Sentence denotes 2.7.
T94 8077-8081 Sentence denotes 2.7.
T4053 8082-8123 Sentence denotes Enzyme-Linked Immunosorbent Assay (ELISA)
T95 8082-8123 Sentence denotes Enzyme-Linked Immunosorbent Assay (ELISA)
T4054 8124-8181 Sentence denotes Cells of four groups were homogenized as mentioned above.
T96 8124-8181 Sentence denotes Cells of four groups were homogenized as mentioned above.
T4055 8182-8304 Sentence denotes The supernatant was collected, and total protein was determined by Coomassie Plus Protein Assay Reagent (Pierce, IL, USA).
T97 8182-8304 Sentence denotes The supernatant was collected, and total protein was determined by Coomassie Plus Protein Assay Reagent (Pierce, IL, USA).
T98 8305-8533 Sentence denotes Levels of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6) protein were quantified using ELISA kits specific for mouse according to the manufacturer's instructions (Bender MedSystems Inc.
T4056 8305-8543 Sentence denotes Levels of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6) protein were quantified using ELISA kits specific for mouse according to the manufacturer's instructions (Bender MedSystems Inc. CA, USA).
T99 8534-8543 Sentence denotes CA, USA).
T4057 8544-8637 Sentence denotes Briefly, prepared the standard and created the standard dilution for building standard curve.
T100 8544-8637 Sentence denotes Briefly, prepared the standard and created the standard dilution for building standard curve.
T4058 8638-8702 Sentence denotes Then samples and biotinconjugate were added to microwell strips.
T101 8638-8702 Sentence denotes Then samples and biotinconjugate were added to microwell strips.
T4059 8703-8852 Sentence denotes After incubated for 2 h at room temperature, the microwell strips were washed 3 times with wash buffer, and streptavidin-HRP were added to all wells.
T102 8703-8852 Sentence denotes After incubated for 2 h at room temperature, the microwell strips were washed 3 times with wash buffer, and streptavidin-HRP were added to all wells.
T4060 8853-8948 Sentence denotes After incubated for 1 h, microwell strips were washed 3 times followed by adding TMB substrate.
T103 8853-8948 Sentence denotes After incubated for 1 h, microwell strips were washed 3 times followed by adding TMB substrate.
T4061 8949-9014 Sentence denotes After incubated for about 10–30 min, the stop solution was added.
T104 8949-9014 Sentence denotes After incubated for about 10–30 min, the stop solution was added.
T4062 9015-9131 Sentence denotes The colour intensity was measured at 450 nm using a Bio-Rad ELISA microplate reader (Bio-Rad Laboratories, CA, USA).
T105 9015-9131 Sentence denotes The colour intensity was measured at 450 nm using a Bio-Rad ELISA microplate reader (Bio-Rad Laboratories, CA, USA).
T4063 9132-9248 Sentence denotes The concentration of protein was determined according to the standard curve and expressed as pg/mg of total protein.
T106 9132-9248 Sentence denotes The concentration of protein was determined according to the standard curve and expressed as pg/mg of total protein.
T4378 9250-9254 Sentence denotes 2.8.
T107 9250-9254 Sentence denotes 2.8.
T4379 9255-9274 Sentence denotes Gelatine Zymography
T108 9255-9274 Sentence denotes Gelatine Zymography
T109 9275-9420 Sentence denotes Cells of four groups were homogenized in lysis buffer containing 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 5 mM CaCl2, 0.2 mM NaN3, and 0.01% Triton.
T4380 9275-9604 Sentence denotes Cells of four groups were homogenized in lysis buffer containing 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 5 mM CaCl2, 0.2 mM NaN3, and 0.01% Triton. Soluble extracts were separated by centrifugation and stored at –20°C. Gelatin zymography was performed according to the manufacturer's instructions (Genmed Scientifics Inc, MA, USA).
T110 9421-9491 Sentence denotes Soluble extracts were separated by centrifugation and stored at –20°C.
T111 9492-9604 Sentence denotes Gelatin zymography was performed according to the manufacturer's instructions (Genmed Scientifics Inc, MA, USA).
T4381 9605-9681 Sentence denotes Briefly, 40 μg cytosolic protein extracts were separated by electrophoresis.
T112 9605-9681 Sentence denotes Briefly, 40 μg cytosolic protein extracts were separated by electrophoresis.
T4382 9682-9847 Sentence denotes Then the proteins were renatured by incubation in 2.5% Triton X-100 and then incubated in substrate buffer for 40 h at 37°C to enable the MMP9 to cleave the gelatin.
T113 9682-9847 Sentence denotes Then the proteins were renatured by incubation in 2.5% Triton X-100 and then incubated in substrate buffer for 40 h at 37°C to enable the MMP9 to cleave the gelatin.
T4383 9848-9951 Sentence denotes After rinsing in water, each gel was stained with Coomassie blue for 1 h and destained in 50% methanol.
T114 9848-9951 Sentence denotes After rinsing in water, each gel was stained with Coomassie blue for 1 h and destained in 50% methanol.
T4384 9952-10057 Sentence denotes Proteolytic activities were showed by clear bands in blue gel which indicates the lysis of the substrate.
T115 9952-10057 Sentence denotes Proteolytic activities were showed by clear bands in blue gel which indicates the lysis of the substrate.
T4385 10058-10143 Sentence denotes Quantification of MMP9 band density was performed with image analysis program ImageJ.
T116 10058-10143 Sentence denotes Quantification of MMP9 band density was performed with image analysis program ImageJ.
T4757 10145-10149 Sentence denotes 2.9.
T117 10145-10149 Sentence denotes 2.9.
T4758 10150-10170 Sentence denotes Statistical Analysis
T118 10150-10170 Sentence denotes Statistical Analysis
T4759 10171-10260 Sentence denotes Data were expressed as mean ± SD and evaluated by ANOVA and LSD multiple comparison test.
T119 10171-10260 Sentence denotes Data were expressed as mean ± SD and evaluated by ANOVA and LSD multiple comparison test.
T4760 10261-10310 Sentence denotes P values <0.05 were considered to be significant.
T120 10261-10310 Sentence denotes P values <0.05 were considered to be significant.
T4761 10311-10367 Sentence denotes All analyses were performed by using SPSS 18.0 software.
T121 10311-10367 Sentence denotes All analyses were performed by using SPSS 18.0 software.
T122 10369-10371 Sentence denotes 3.
T123 10372-10379 Sentence denotes Results
T4844 10381-10385 Sentence denotes 3.1.
T124 10381-10385 Sentence denotes 3.1.
T4845 10386-10464 Sentence denotes Depletion of Nrf2 Aggravated the Cell Death in Astrocytes after Scratch Injury
T125 10386-10464 Sentence denotes Depletion of Nrf2 Aggravated the Cell Death in Astrocytes after Scratch Injury
T4846 10465-10544 Sentence denotes Cell death was detected by microscope and CCK-8 analysis at 24 h after scratch.
T126 10465-10544 Sentence denotes Cell death was detected by microscope and CCK-8 analysis at 24 h after scratch.
T4847 10545-10693 Sentence denotes The detachment from culture plate, cell lost, and cytoplasmic process distortion were mostly observed in both sides of scratch line in group KO TBI.
T127 10545-10693 Sentence denotes The detachment from culture plate, cell lost, and cytoplasmic process distortion were mostly observed in both sides of scratch line in group KO TBI.
T4848 10694-10881 Sentence denotes CCK-8 assay also showed lower OD value of group KO TBI (0.38 ± 0.064) than that of group WT TBI (0.98 ± 0.098) (P < 0.01), which also suggested more cell death in group KO TBI (Figure 1).
T128 10694-10881 Sentence denotes CCK-8 assay also showed lower OD value of group KO TBI (0.38 ± 0.064) than that of group WT TBI (0.98 ± 0.098) (P < 0.01), which also suggested more cell death in group KO TBI (Figure 1).
T4849 10882-10939 Sentence denotes There was no difference between two sham-operated groups.
T129 10882-10939 Sentence denotes There was no difference between two sham-operated groups.
T5106 10941-10945 Sentence denotes 3.2.
T130 10941-10945 Sentence denotes 3.2.
T5107 10946-11037 Sentence denotes Disruption of Nrf2 Enhanced Upregulation of NF-κB DNA-Binding Activity after Scratch Injury
T131 10946-11037 Sentence denotes Disruption of Nrf2 Enhanced Upregulation of NF-κB DNA-Binding Activity after Scratch Injury
T5108 11038-11106 Sentence denotes It has been reported that NF-κB is activated in brain after TBI [2].
T132 11038-11106 Sentence denotes It has been reported that NF-κB is activated in brain after TBI [2].
T5109 11107-11233 Sentence denotes Here, we studied the DNA-binding activity of NF-κB of astrocytes from Nrf2 WT or KO mice by EMSA at 24 h after scratch injury.
T133 11107-11233 Sentence denotes Here, we studied the DNA-binding activity of NF-κB of astrocytes from Nrf2 WT or KO mice by EMSA at 24 h after scratch injury.
T5110 11234-11444 Sentence denotes Scratch injury induced activation of NF-κB in astrocytes of both genotypes, while higher NF-κB activity was observed in group KO TBI than in group WT TBI (2.67 ± 0.173 versus 2.28 ± 0.072, P < 0.01) (Figure 2).
T134 11234-11444 Sentence denotes Scratch injury induced activation of NF-κB in astrocytes of both genotypes, while higher NF-κB activity was observed in group KO TBI than in group WT TBI (2.67 ± 0.173 versus 2.28 ± 0.072, P < 0.01) (Figure 2).
T5417 11446-11450 Sentence denotes 3.3.
T135 11446-11450 Sentence denotes 3.3.
T5418 11451-11552 Sentence denotes Expression of Proinflammatory Cytokines Was Elevated in Nrf2 Knockout Astrocytes after Scratch Injury
T136 11451-11552 Sentence denotes Expression of Proinflammatory Cytokines Was Elevated in Nrf2 Knockout Astrocytes after Scratch Injury
T5419 11553-11660 Sentence denotes As we all know, TNF-α, IL-1β, and IL-6 are regulated by NF-κB and reflect the endogenous activity of NF-κB.
T137 11553-11660 Sentence denotes As we all know, TNF-α, IL-1β, and IL-6 are regulated by NF-κB and reflect the endogenous activity of NF-κB.
T5420 11661-11761 Sentence denotes It has been revealed that these proinflammatory cytokines are increased after TBI in animals [2, 8].
T138 11661-11761 Sentence denotes It has been revealed that these proinflammatory cytokines are increased after TBI in animals [2, 8].
T5421 11762-11913 Sentence denotes And our previous study has proved that higher levels of such proinflammatory cytokines were observed in the brain of Nrf2-deficient mice after TBI [5].
T139 11762-11913 Sentence denotes And our previous study has proved that higher levels of such proinflammatory cytokines were observed in the brain of Nrf2-deficient mice after TBI [5].
T5422 11914-12020 Sentence denotes But it is still ambiguous about the relationship between these cytokines and Nrf2 in astrocytes after TBI.
T140 11914-12020 Sentence denotes But it is still ambiguous about the relationship between these cytokines and Nrf2 in astrocytes after TBI.
T5423 12021-12146 Sentence denotes Here, we tested the levels of TNF-α, IL-1β, and IL-6 in cultured astrocytes at 24 h after scratch injury by RT-PCR and ELISA.
T141 12021-12146 Sentence denotes Here, we tested the levels of TNF-α, IL-1β, and IL-6 in cultured astrocytes at 24 h after scratch injury by RT-PCR and ELISA.
T5424 12147-12322 Sentence denotes It was shown that the mRNA levels of TNF-α, IL-1β, and IL-6 were increased after scratch in both WT and KO astrocytes when compared with their sham counterparts, respectively.
T142 12147-12322 Sentence denotes It was shown that the mRNA levels of TNF-α, IL-1β, and IL-6 were increased after scratch in both WT and KO astrocytes when compared with their sham counterparts, respectively.
T5425 12323-12563 Sentence denotes Moreover, the mRNA levels of these cytokines in group KO TBI were much higher than those in group WT TBI (1.66 ± 0.085 versus 1.39 ± 0.110 for TNF-α, 1.34 ± 0.064 versus 0.73 ± 0.088 for IL-1β, and 1.28 ± 0.102 versus 0.99 ± 0.073 for IL-6.
T143 12323-12563 Sentence denotes Moreover, the mRNA levels of these cytokines in group KO TBI were much higher than those in group WT TBI (1.66 ± 0.085 versus 1.39 ± 0.110 for TNF-α, 1.34 ± 0.064 versus 0.73 ± 0.088 for IL-1β, and 1.28 ± 0.102 versus 0.99 ± 0.073 for IL-6.
T5426 12564-12594 Sentence denotes P < 0.01) (Figures 3(a)–3(d)).
T144 12564-12594 Sentence denotes P < 0.01) (Figures 3(a)–3(d)).
T5427 12595-12761 Sentence denotes The results of ELISA revealed that the protein level of TNF-α was upregulated after scratch injury in WT and KO astrocytes when compared with their sham counterparts.
T145 12595-12761 Sentence denotes The results of ELISA revealed that the protein level of TNF-α was upregulated after scratch injury in WT and KO astrocytes when compared with their sham counterparts.
T5428 12762-12881 Sentence denotes And it was also higher in group KO TBI than in group WT TBI (3.67 ± 0.156 versus 2.31 ± 0.087, P < 0.01) (Figure 4(a)).
T146 12762-12881 Sentence denotes And it was also higher in group KO TBI than in group WT TBI (3.67 ± 0.156 versus 2.31 ± 0.087, P < 0.01) (Figure 4(a)).
T5429 12882-12947 Sentence denotes Similar tendency was observed in protein level of IL-6 and IL-1β.
T147 12882-12947 Sentence denotes Similar tendency was observed in protein level of IL-6 and IL-1β.
T5430 12948-13058 Sentence denotes For IL-6, it was 36.07 ± 0.786 for group KO TBI while 25.76 ± 0.536 for group WT TBI (P < 0.01) (Figure 4(b)).
T148 12948-13058 Sentence denotes For IL-6, it was 36.07 ± 0.786 for group KO TBI while 25.76 ± 0.536 for group WT TBI (P < 0.01) (Figure 4(b)).
T5431 13059-13172 Sentence denotes For IL-1β, it was 190.75 ± 6.339 for group KO TBI while 154.50 ± 5.348 for group WT TBI (P < 0.01) (Figure 4(c)).
T149 13059-13172 Sentence denotes For IL-1β, it was 190.75 ± 6.339 for group KO TBI while 154.50 ± 5.348 for group WT TBI (P < 0.01) (Figure 4(c)).
T6473 13174-13178 Sentence denotes 3.4.
T150 13174-13178 Sentence denotes 3.4.
T6474 13179-13281 Sentence denotes Expression and Activity of MMP9 Were Greatly Enhanced in Nrf2 Knockout Astrocytes after Scratch Injury
T151 13179-13281 Sentence denotes Expression and Activity of MMP9 Were Greatly Enhanced in Nrf2 Knockout Astrocytes after Scratch Injury
T6475 13282-13362 Sentence denotes MMP9 is an important gelatinase to induce or aggravate the inflammation process.
T152 13282-13362 Sentence denotes MMP9 is an important gelatinase to induce or aggravate the inflammation process.
T6476 13363-13536 Sentence denotes In the present study, we elevated the mRNA, protein levels, and activity of MMP9 in astrocytes at 24 h after scratch injury by RT-PCR, western blot, and gelatine zymography.
T153 13363-13536 Sentence denotes In the present study, we elevated the mRNA, protein levels, and activity of MMP9 in astrocytes at 24 h after scratch injury by RT-PCR, western blot, and gelatine zymography.
T6477 13537-13655 Sentence denotes Expression of MMP9 was elevated after scratch injury in WT and KO astrocytes as compared with their sham counterparts.
T154 13537-13655 Sentence denotes Expression of MMP9 was elevated after scratch injury in WT and KO astrocytes as compared with their sham counterparts.
T6478 13656-13796 Sentence denotes The mRNA level of MMP9 was higher in group KO TBI than in group WT TBI (1.36 ± 0.090 versus 1.01 ± 0.068, P < 0.01) (Figures 3(a) and 3(e)).
T155 13656-13796 Sentence denotes The mRNA level of MMP9 was higher in group KO TBI than in group WT TBI (1.36 ± 0.090 versus 1.01 ± 0.068, P < 0.01) (Figures 3(a) and 3(e)).
T6479 13797-13968 Sentence denotes Significant difference was also discovered in MMP9 protein level by western blot, as 1.44 ± 0.076 for group KO TBI and 1.13 ± 0.048 for group WT TBI (P < 0.01) (Figure 5).
T156 13797-13968 Sentence denotes Significant difference was also discovered in MMP9 protein level by western blot, as 1.44 ± 0.076 for group KO TBI and 1.13 ± 0.048 for group WT TBI (P < 0.01) (Figure 5).
T6480 13969-14124 Sentence denotes Gelatine zymography revealed higher MMP9 activity in group KO TBI than that in group WT TBI too (101.76 ± 6.343 versus 76.32 ± 3.388, P < 0.01) (Figure 6).
T157 13969-14124 Sentence denotes Gelatine zymography revealed higher MMP9 activity in group KO TBI than that in group WT TBI too (101.76 ± 6.343 versus 76.32 ± 3.388, P < 0.01) (Figure 6).
T158 14126-14128 Sentence denotes 4.
T159 14129-14139 Sentence denotes Discussion
T7214 14140-14320 Sentence denotes The present study demonstrated that scratch injury induced the upregulation of NF-κB DNA binding activity and overexpression of TNF-α, IL-1β, IL-6, and MMP9 in cultured astrocytes.
T160 14140-14320 Sentence denotes The present study demonstrated that scratch injury induced the upregulation of NF-κB DNA binding activity and overexpression of TNF-α, IL-1β, IL-6, and MMP9 in cultured astrocytes.
T7215 14321-14534 Sentence denotes Also we revealed that scratch injury induced higher activity of NF-κB and enhanced expression of proinflammatory cytokines in Nrf2 knockout cultured astrocytes than those in wildtype astrocytes for the first time.
T161 14321-14534 Sentence denotes Also we revealed that scratch injury induced higher activity of NF-κB and enhanced expression of proinflammatory cytokines in Nrf2 knockout cultured astrocytes than those in wildtype astrocytes for the first time.
T7216 14535-14639 Sentence denotes It has been demonstrated that NF-κB is activated in brain and spinal cord after traumatic injury [2, 9].
T162 14535-14639 Sentence denotes It has been demonstrated that NF-κB is activated in brain and spinal cord after traumatic injury [2, 9].
T7217 14640-14858 Sentence denotes As a transcript factor, NF-κB binds with DNA once it was activated and induces transcription of MMP9 and a battery of proinflammatory cytokines, including TNF-α, IL-1β, and IL-6, in brain tissues after TBI [3, 10, 11].
T163 14640-14858 Sentence denotes As a transcript factor, NF-κB binds with DNA once it was activated and induces transcription of MMP9 and a battery of proinflammatory cytokines, including TNF-α, IL-1β, and IL-6, in brain tissues after TBI [3, 10, 11].
T7218 14859-15047 Sentence denotes Our results also revealed that scratch injury induced elevation of NF-κB DNA binding activity, overexpression of MMP9, and proinflammatory mediators mentioned above in cultured astrocytes.
T164 14859-15047 Sentence denotes Our results also revealed that scratch injury induced elevation of NF-κB DNA binding activity, overexpression of MMP9, and proinflammatory mediators mentioned above in cultured astrocytes.
T7219 15048-15148 Sentence denotes It has been demonstrated that there is an autoregulatory loop among these proinflammatory mediators.
T165 15048-15148 Sentence denotes It has been demonstrated that there is an autoregulatory loop among these proinflammatory mediators.
T7220 15149-15376 Sentence denotes For example, TNF-α and IL-1β are potent stimulators for MMP9 in astrocytes [12, 13], and IL-6 induces overexpression of MMP9 in human colon carcinoma cells [14], and TNF-α and IL-1β also can induce activation of NF-κB [15, 16].
T166 15149-15376 Sentence denotes For example, TNF-α and IL-1β are potent stimulators for MMP9 in astrocytes [12, 13], and IL-6 induces overexpression of MMP9 in human colon carcinoma cells [14], and TNF-α and IL-1β also can induce activation of NF-κB [15, 16].
T7221 15377-15497 Sentence denotes This autoregulatory loop extremely aggravates the damaging effect of inflammation and induces secondary injury to brain.
T167 15377-15497 Sentence denotes This autoregulatory loop extremely aggravates the damaging effect of inflammation and induces secondary injury to brain.
T7222 15498-15600 Sentence denotes It is a good choice for targeting on an upstream factor to prevent such autoregulatory loop after TBI.
T168 15498-15600 Sentence denotes It is a good choice for targeting on an upstream factor to prevent such autoregulatory loop after TBI.
T7223 15601-15778 Sentence denotes Nrf2-ARE pathway has been proved to be the key regulator in reducing oxidative stress, inflammatory damage, and accumulation of toxic metabolites, which are all involved in TBI.
T169 15601-15778 Sentence denotes Nrf2-ARE pathway has been proved to be the key regulator in reducing oxidative stress, inflammatory damage, and accumulation of toxic metabolites, which are all involved in TBI.
T7224 15779-15865 Sentence denotes Our previous study has proved the augmentation of Nrf2 in brain tissue after TBI [17].
T170 15779-15865 Sentence denotes Our previous study has proved the augmentation of Nrf2 in brain tissue after TBI [17].
T7225 15866-16082 Sentence denotes Enhanced level of Nrf2 activates transcription of a group of antioxidant genes, such as heme oxygenase-1 (HO-1) and NAD (P)H: quinone oxidoreductase-1 (NQO1), which would subsequently reduce the damage in brain [18].
T171 15866-16082 Sentence denotes Enhanced level of Nrf2 activates transcription of a group of antioxidant genes, such as heme oxygenase-1 (HO-1) and NAD (P)H: quinone oxidoreductase-1 (NQO1), which would subsequently reduce the damage in brain [18].
T7226 16083-16344 Sentence denotes Postinjury administration of SFN, an inducer of Nrf2, significantly improves spatial memory of rat and decreases the immunoreactivity for 4-Hydroxynonenal (4-HNE), a marker of lipid peroxidation, in the cortex and the CA3 subfield of hippocampus after TBI [19].
T172 16083-16344 Sentence denotes Postinjury administration of SFN, an inducer of Nrf2, significantly improves spatial memory of rat and decreases the immunoreactivity for 4-Hydroxynonenal (4-HNE), a marker of lipid peroxidation, in the cortex and the CA3 subfield of hippocampus after TBI [19].
T7227 16345-16415 Sentence denotes On the other hand, Nrf2-deficient mice appear more susceptible to TBI.
T173 16345-16415 Sentence denotes On the other hand, Nrf2-deficient mice appear more susceptible to TBI.
T7228 16416-16513 Sentence denotes Depletion of Nrf2 induces higher expression of proinflammatory mediators in brain after TBI [20].
T174 16416-16513 Sentence denotes Depletion of Nrf2 induces higher expression of proinflammatory mediators in brain after TBI [20].
T7229 16514-16804 Sentence denotes Here, our results revealed that overexpression of TNF-α, IL-1β, IL-6, and MMP9 after scratch injury was more aggravated in cultured Nrf2 knockout astrocytes than in wildtype astrocyetes for the first time, and overexpression of these proinflammatory mediators led to more astrocytes deaths.
T175 16514-16804 Sentence denotes Here, our results revealed that overexpression of TNF-α, IL-1β, IL-6, and MMP9 after scratch injury was more aggravated in cultured Nrf2 knockout astrocytes than in wildtype astrocyetes for the first time, and overexpression of these proinflammatory mediators led to more astrocytes deaths.
T7230 16805-16992 Sentence denotes Data obtained from animal studies suggest the possibility that antioxidant effect of Nrf2 may be achieved by suppression of proinflammatory pathways which are mediated by NF-κB signaling.
T176 16805-16992 Sentence denotes Data obtained from animal studies suggest the possibility that antioxidant effect of Nrf2 may be achieved by suppression of proinflammatory pathways which are mediated by NF-κB signaling.
T7231 16993-17161 Sentence denotes Administration of SFN is found to be able to inhibit IKK/IκB phosphorylation and p65 NF-κB subunit nuclear translocation, consequently alleviating NF-κB signaling [21].
T177 16993-17161 Sentence denotes Administration of SFN is found to be able to inhibit IKK/IκB phosphorylation and p65 NF-κB subunit nuclear translocation, consequently alleviating NF-κB signaling [21].
T7232 17162-17303 Sentence denotes And NF-κB activation induced by lipopolysaccharide (LPS) could be attenuated by diverse Nrf2 activators, such as SFN and curcumin (CUR) [22].
T178 17162-17303 Sentence denotes And NF-κB activation induced by lipopolysaccharide (LPS) could be attenuated by diverse Nrf2 activators, such as SFN and curcumin (CUR) [22].
T7233 17304-17486 Sentence denotes Furthermore, our previous studies indicate that depletion of Nrf2 induces augmentation of NF-κB activity and inflammatory response in lung, brain, and intestine after TBI [4, 5, 23].
T179 17304-17486 Sentence denotes Furthermore, our previous studies indicate that depletion of Nrf2 induces augmentation of NF-κB activity and inflammatory response in lung, brain, and intestine after TBI [4, 5, 23].
T7234 17487-17599 Sentence denotes Results from this study further confirmed such relationship existed in cultured astrocytes after scratch injury.
T180 17487-17599 Sentence denotes Results from this study further confirmed such relationship existed in cultured astrocytes after scratch injury.
T7235 17600-17756 Sentence denotes Enhanced activation of NF-κB is also discovered in lung, macrophages, and mouse embryonic fibroblasts of Nrf2-deficient mice after experimental sepsis [24].
T181 17600-17756 Sentence denotes Enhanced activation of NF-κB is also discovered in lung, macrophages, and mouse embryonic fibroblasts of Nrf2-deficient mice after experimental sepsis [24].
T7236 17757-17840 Sentence denotes Interestingly, it is reported that NF-κB can inhibit Nrf2 at transcriptional level.
T182 17757-17840 Sentence denotes Interestingly, it is reported that NF-κB can inhibit Nrf2 at transcriptional level.
T7237 17841-18011 Sentence denotes NF-κB p65 subunit repressed the Nrf2-ARE pathway at transcriptional level by competitive interaction with the CH1-KIX domain of CBP or local histone hypoacetylation [25].
T183 17841-18011 Sentence denotes NF-κB p65 subunit repressed the Nrf2-ARE pathway at transcriptional level by competitive interaction with the CH1-KIX domain of CBP or local histone hypoacetylation [25].
T7238 18012-18192 Sentence denotes All these findings indicate the potential complicate crosstalk between NF-κB and Nrf2, which may be regulated by the upstream mitogen-activated protein kinase (MAPKs) pathway [26].
T184 18012-18192 Sentence denotes All these findings indicate the potential complicate crosstalk between NF-κB and Nrf2, which may be regulated by the upstream mitogen-activated protein kinase (MAPKs) pathway [26].
T7239 18193-18284 Sentence denotes It has been confirmed that Nrf2 is mainly detected in nucleus of astrocytes after TBI [18].
T185 18193-18284 Sentence denotes It has been confirmed that Nrf2 is mainly detected in nucleus of astrocytes after TBI [18].
T7240 18285-18502 Sentence denotes In view of the fact that Nrf2 is a transcription factor which should take function mainly in nucleus, it can be reasoned that astrocytes may be one kind of respondent cells in activation of Nrf2-ARE pathway after TBI.
T186 18285-18502 Sentence denotes In view of the fact that Nrf2 is a transcription factor which should take function mainly in nucleus, it can be reasoned that astrocytes may be one kind of respondent cells in activation of Nrf2-ARE pathway after TBI.
T7241 18503-18644 Sentence denotes Previous study identified the expression of TNF-α, IL-1β, and IL-6 in cultured astrocytes after treatment with LPS or oxyhemoglobin [27, 28].
T187 18503-18644 Sentence denotes Previous study identified the expression of TNF-α, IL-1β, and IL-6 in cultured astrocytes after treatment with LPS or oxyhemoglobin [27, 28].
T7242 18645-18798 Sentence denotes Another study revealed that after middle cerebral artery occlusion, MMP9-positive astrocytes were observed in brain tissues by immunohistochemistry [29].
T188 18645-18798 Sentence denotes Another study revealed that after middle cerebral artery occlusion, MMP9-positive astrocytes were observed in brain tissues by immunohistochemistry [29].
T7243 18799-18888 Sentence denotes Those results indicate the role of astrocytes in inflammatory process after brain injury.
T189 18799-18888 Sentence denotes Those results indicate the role of astrocytes in inflammatory process after brain injury.
T7244 18889-19013 Sentence denotes But till now, there is no study focused on the relationship among astrocytes, Nrf2, and proinflammatory mediators after TBI.
T190 18889-19013 Sentence denotes But till now, there is no study focused on the relationship among astrocytes, Nrf2, and proinflammatory mediators after TBI.
T7245 19014-19166 Sentence denotes Our results demonstrated the upregulated expression of TNF-α, IL-1β, IL-6, and MMP9 in Nrf2 knockout astrocytes after scratch injury for the first time.
T191 19014-19166 Sentence denotes Our results demonstrated the upregulated expression of TNF-α, IL-1β, IL-6, and MMP9 in Nrf2 knockout astrocytes after scratch injury for the first time.
T7246 19167-19348 Sentence denotes In conclusion, depletion of Nrf2 induced the activation of NF-κB and the expression of TNF-α, IL-1β, IL-6, and MMP9 resulting in more cell deaths in astrocytes after scratch injury.
T192 19167-19348 Sentence denotes In conclusion, depletion of Nrf2 induced the activation of NF-κB and the expression of TNF-α, IL-1β, IL-6, and MMP9 resulting in more cell deaths in astrocytes after scratch injury.
T7247 19349-19448 Sentence denotes These results suggest that Nrf2 may be an important target for anti-inflammatory therapy after TBI.
T193 19349-19448 Sentence denotes These results suggest that Nrf2 may be an important target for anti-inflammatory therapy after TBI.