中医药学报2024，Vol.52Issue(5)：30-37,8.DOI:10.19664/j.cnki.1002-2392.240091
基于氧化应激探讨疏肝补肾法对卵巢早衰大鼠颗粒细胞凋亡的影响
Effect of Soothing Liver and Tonifying Kidney on Apoptosis of Granulosa Cells in Rats with Premature Ovarian Failure based on Oxidative Stress
摘要
Abstract
Objective:To investigate the possible mechanism of soothing the liver and tonifying the kidney on the apoptosis of granulosa cells in rats with premature ovarian failure.Methods:Forty-eight SD rats were randomly divided into normal group,model group,bupleurum liver-soothing powder plus left-restoring beverage(BLP + LRB)low dose,medium dose,and high dose group,and estrogen group,with 8 rats in each group.Except for the normal group,the rats in other groups were intraperitoneally injected cyclophosphamide for 15 days to establish a model of premature ovarian failure,while the chronic unpredictable mild stress mode was used for 21 days to establish a model of liver depression.After successful modeling,BLP +LRB low,medium and high dose groups were administrated with BLP + LRB suspension 3.39 g·kg-1,6.77 g·kg-1 and 13.55 g·kg-1,respectively,the estrogen group was administrated with estradiol valerate(bujiale)0.105 mg·kg-1,and the normal group and model group were administrated with equal volume of sterilized water for 3 weeks.Exfoliated vaginal cells were collected and the changes of estrous cycle in rats were monitored.The modeling of liver depression syndrome was detected by behavior.The histopathology of the ovary was observed by HE staining,the contents of serum estradiol(E2),follicle stimulating hormone(FSH),anti-Muller-tube hormone(AMH),and Inhibin B(INHB)were detected by ELISA,and the apoptosis index of ovarian granulosa cells was detected by TUNEL method.The mRNA and protein expressions of Bcl-2 Associated X gene(Bax),B-cell lymphoma-2 gene(Bcl-2),reactive oxygen species(ROS)and superoxide dismutase-2(SOD-2)in ovarian tissue were detected by q-PCR and Western Blot.Results:Compared with the normal group,the estrous cycle of rats in the model group was in disorder,the ovarian tissue structure was unclear,the granulosa cell layer was obviously thinner,the total distance of open field experiment movement and sucrose water consumption rate decreased(P<0.05,P<0.01),the serum FSH level increased,the levels of E2,AMH and INHB were significantly decreased(P<0.01),and the granulosa cell apoptosis index increased(P<0.01).The mRNA and protein expressions of Bcl-2 and SOD-2 decreased,the mRNA and protein expressions of Bax increased,and the mRNA and protein expressions of ROS increased(P<0.05,P<0.01).Compared with the model group,the experimental results of pathological morphology of ovarian tissue and behavior in all drug administration groups were improved,serum FSH level decreased,E2 and INHB levels increased(P<0.05,P<0.01),mRNA expression levels of Bax and ROS decreased.Granulosa cell apoptosis index decreased(P<0.01)and Bcl-2 protein expression level increased in BLP + LRB medium and high dose groups and estrogen group.The expression of Bax protein decreased and the mRNA and protein expression of SOD-2 and Bcl-2 increased in BLP +LRB high-dose group and estrogen group(P<0.05,P<0.01).Compared with BLP +LRB low dose group,INHB level in BLP +LRB medium dose group increased,FSH decreased,but E2 in BLP +LRB high dose group increased,SOD-2 protein expression increased in BLP + LRB high dose group,and the apoptosis index of granulosa cells in BLP + LRB medium and high dose groups decreased(P<0.05,P<0.01).Compared with BLP +LRB medium dose group,E2 level increased and Bax mRNA expression level decreased in BLP + LRB high dose group(P<0.05).Conclusion:Soothing the liver and tonifying the kidney may reduce the apoptosis rate of granulosa cells and improve ovarian function by alleviating oxidative stress,and the effect of high dose group is the best.关键词
卵巢早衰/氧化应激/疏肝补肾法/颗粒细胞凋亡Key words
Premature ovarian failure/Oxidative stress/Soothing the liver and tonifying the kidney/Granulosa cell apoptosis分类
医药卫生引用本文复制引用
王敏,张斐,王洋..基于氧化应激探讨疏肝补肾法对卵巢早衰大鼠颗粒细胞凋亡的影响[J].中医药学报,2024,52(5):30-37,8.基金项目
国家自然科学基金青年科学基金项目(82004257) （82004257）
福建省自然科学基金项目(2021J01933) （2021J01933）
