HONG Yan-li, TAN Yong. Effect of Yiqi Yangyin Formula on Mitochondrial Apoptotic Pathway of Ovarian Granulosa Cells Induced by cDDP[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(6): 104-108.
DOI:
HONG Yan-li, TAN Yong. Effect of Yiqi Yangyin Formula on Mitochondrial Apoptotic Pathway of Ovarian Granulosa Cells Induced by cDDP[J]. Chinese journal of experimental traditional medical formulae, 2016, 22(6): 104-108. DOI: 10.13422/j.cnki.syfjx.2016060104.
Effect of Yiqi Yangyin Formula on Mitochondrial Apoptotic Pathway of Ovarian Granulosa Cells Induced by cDDP
Objective: To discuss the mechanism of Yiqi Yangyin formula on mitochondrial pathway of granulosa cells apoptosis induced by cDDP. Method: Granulosa cells apoptosis model was induced by cDDP. Then drug-containing serum of different concentrations (10.5
21
31.5 g·kg-1) were used to intervene in apoptosis. The blank group was also set up. The proliferation and apoptosis were analyzed by MTT assays and flowcytometry. GC apoptosis rate was detected with TUNEL staining. Calcium concentration and mitochondrial membrane potential were detected by confocal laser scanning microscope. The mRNA expressions of Bcl-2
Bad
Caspases-3 and Caspases-9 were detected by RT-PCR;protein levels of PI3K
Akt and p-Akt were detected by western blot
respectively. Result: Compared with the blank group
the model group showed significant increase in GC apoptosis rate and Ca2+ fluorescence intensity and decrease in mitochondrial membrane potential(MMP). Besides
the mRNA expressions of Bad
Caspases-3
Caspases-9 were significantly higher in model group than those in control group (P<0.05). Meanwhile
Bcl-2 mRNA expression
PI3K and p-Akt were significantly lower in model group (P<0.05). In middle and high dose groups
calcium concentration
the mRNA expression of Bcl-2
Caspase-3 and Caspase-9 were down-regulated significantly (P<0.05)
while the mitochondrial membrane potential
Bcl-2 mRNA expression
and the protein levels of PI3K and p-Akt were up-regulated significantly (P<0.05). Conclusion: Yiqi Yangyin formula can inhibit GC apoptosis induced by cDDP by activating PI3K/Akt signal pathway
up-regulating anti-apoptosis factors (Bcl-2)
down-regulating expressions of pro-apoptosis factors (Bad and caspases family)
stabilizing the mitochondrial membrane potential and reducing calcium influx.