PMC:7391798 / 8991-12599 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/7391798","sourcedb":"PMC","sourceid":"7391798","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7391798","text":"Results\n\nImpact of myoglobin on proliferation and apoptosis\nAs the concentration of myoglobin increased, the inhibition of NRK-52E’s cell viability gradually increased, and the apoptosis rate of NRK-52E cells increased. Early apoptosis of NRK-52E increased in group 100, and late apoptosis of NRK-52E increased in group 150. Late apoptosis often indicates that cells begin to show more irreversible damage and eventually evolve into cell death, Figure 1.\n\nResults of autophagy PCR chip screening\nA total of 96 autophagy-related genes were screened by the autophagy PCR chip. The results showed that the highest expression of Pink1 was in group 100, but the expression in group 150 was lower than other groups. In addition, group 100 had the lowest Parkin expression. Beclin1 was expressed at a higher level, and these 3 genes were all on the Pink1/Parkin autophagy pathway by the KEGG PATHWAY database analysis (Table 2, Figure 2).\n\nImpact of myoglobin on expressions of Pink1, Beclin1, and Parkin mRNA\nIn group 50, group 100, and group 150, NRK-52E expressed higher Pink1 mRNA than that in group 0, but the expressions of Parkin mRNA were lower than in group 0. The expressions of Parkin mRNA in group 100 and group 150 were lower than in group 0. The expressions of Beclin1 mRNA in group 50, group 100, and group 150 were higher than that in group 0, and the expressions of Beclin1 mRNA in group 100 and group 150 were higher than in group 50 (Figure 3).\n\nImpact of myoglobin on expressions of Pink1/Parkin and autophagy-related proteins\nAfter adding myoglobin to NRK-52E, the expression of Parkin gradually decreased, but the expression of Pink1 gradually increased; the expression of P62 decreased; and the ratio of LC3II/LC3I increased. When the final myoglobin concentration was 150 μm, the expression increase was the most obvious; the expression of ATG5 gradually increased, and the expression of Beclin1 increased. When the final concentration of myoglobin was 50 μm, the expressions increased the most significantly. There were no significant differences between group 150 and group 100, so we chose 100 μm for further experiments (Figure 4).\n\nImpact of autophagy promoter Rapamycin and Pink1-siRNA on apoptosis\nAfter adding rapamycin and myoglobin to NRK-52E cells, the apoptotic rate increased. However, after being infected with Pink1-siRNA, the apoptotic rate decreased, and the apoptotic rate in Pink1-siRNA-infected groups showed a downward trend (Figure 5).\n\nImpact of autophagy promoter Rapamycin and Pink1-siRNA on expressions of Pink1/Parkin autophagy-related mRNAs and proteins\nCompared with group CON, the expressions of Pink1 mRNA and protein increased in group RAP and group MYO, but the amounts of Pink1 mRNA and protein decreased in group RAP+Pin and group MYO+Pin. The expressions of Parkin mRNA and protein decreased in group RAP and group MYO, and increased in group RAP+Pin and group MYO+Pin. The expression levels of Beclin1 and ATG5 mRNAs and proteins were increased in group RAP and group MYO, but the expressions in group RAP+Pin and group MYO+Pin decreased. The expression of P62 mRNA was the opposite, which decreased in group RAP and group MYO and increased in group RAP+Pin and group MYO+Pin. The ratio of LC3II/LC3I increased in group RAP and group MYP but decreased after Pink1-siRNA infection (Figure 6).\n\nComparison of fluorescent spots of NRK-52E after LC3 double-labeled lentivirus infection\nThe autophagy spots were higher in group RAP and group MYO than in group CON, but the autophagy spots decreased in group RAP+Pin than in group RAP and group MYO (Figure 7).","divisions":[{"label":"title","span":{"begin":0,"end":7}},{"label":"sec","span":{"begin":9,"end":454}},{"label":"title","span":{"begin":9,"end":59}},{"label":"p","span":{"begin":60,"end":454}},{"label":"sec","span":{"begin":456,"end":931}},{"label":"title","span":{"begin":456,"end":495}},{"label":"p","span":{"begin":496,"end":931}},{"label":"sec","span":{"begin":933,"end":1456}},{"label":"title","span":{"begin":933,"end":1002}},{"label":"p","span":{"begin":1003,"end":1456}},{"label":"sec","span":{"begin":1458,"end":2152}},{"label":"title","span":{"begin":1458,"end":1539}},{"label":"p","span":{"begin":1540,"end":2152}},{"label":"sec","span":{"begin":2154,"end":2474}},{"label":"title","span":{"begin":2154,"end":2221}},{"label":"p","span":{"begin":2222,"end":2474}},{"label":"sec","span":{"begin":2476,"end":3345}},{"label":"title","span":{"begin":2476,"end":2598}},{"label":"p","span":{"begin":2599,"end":3345}},{"label":"title","span":{"begin":3347,"end":3435}}],"tracks":[]}