Ameliorative effects of Zigui Yichong Formula on mitochondrial energy metabolism of ovarian granulosa cells via regulation of SIRT1/PGC-1α signaling pathway
AIM To explore the ameliorative effects of Zigui Yichong Formula on mitochondrial energy metabolism in in vitro model of premature ovarian insufficiency (POI).METHODS A POI model in vitro was established in human ovarian granulosa cell KGN with acrolein ( ACR ) for the further intervention of serum containing Zigui Yichong Formula.The control group,the ACR group,the Zigui Yichong Formula group,the EX527 ( SIRT1 inhibitor) group and the EX527+Zigui Yichong Formula group were set up.The cells had their apoptosis rate detected by flow cytometry;their ROS level detected by DCFH-DA method;their mitochondria number detected by Mito-Trakine fluorescence labeling method;their oxidative phosphorylation level of mitochondria detected by Seahorse method;their ATP level detected by colorimetry;their copy number of mtDNA and mRNA expressions of SIRT1,PGC-1α,Nrf1 and TFAM detected by RT-qPCR;and their protein expressions of SIRT1,PGC-1α,Ac-PGC-1α,Nrf1 and TFAM detected by Western blot.RESULTS Compared with the control group,the ACR group displayed increased apoptosis rate and ROS level of KGN cells ( P<0.05,P<0.01 );decreased number of mitochondria and mtDNA copy number ( P<0.01 );decreased basic value of mitochondrial aerobic respiration,maximum respiration,reserve value and ATP level (P<0.05,P<0.01),decreased mRNA and protein expressions of SIRT1,PGC-1α,Nrf1 and TFAM ( P<0.05,P<0.01 );and increased protein expression of Ac-PGC-1α( P<0.01 ).Compared with the ACR group,the Zigui Yichong Formula group demonstrated improvement in terms of the aforementioned indices levels ( P<0.05,P<0.01).The combination use of Zigui Yichong Formula and EX527 reversed the effects of single use of Zigui Yichong Formula on KGN cells ( P<0.05,P<0.01).CONCLUSION Zigui Yichong Formula may improve the mitochondrial energy metabolism disorder induced by ACR in KGN cells by increasing mitochondrial biosynthesis via SIRT1/PGC-1α signaling pathway.
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维普
