摘要
目的:探讨益肾疏肝汤含药血清对雷公藤多苷干预的大鼠颗粒细胞凋亡的影响及其可能的作用机制。方法:分离大鼠卵巢颗粒细胞,使用不同含量的肾疏肝汤含药血清以及不同浓度的雷公藤多苷含药血清处理细胞48 h后,使用CCK-8法检测细胞存活率,筛选益肾疏肝汤含药血清和雷公藤多苷含药血清剂量。并将细胞分为正常组(正常培养的细胞)、损伤组、中药低剂量组、中药中剂量组、中药高剂量组、阳性对照组(雌二醇)。分别采用CCK-8法、TUNEL法、蛋白质印迹法检测细胞增殖、细胞凋亡、凋亡及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路相关蛋白表达。结果:与正常组比较,损伤组细胞增殖、B细胞淋巴瘤-2(Bcl-2)、p-PI3K、p-AKT、p-mTOR蛋白表达均显著受到抑制(P<0.05),而细胞凋亡率、B细胞淋巴瘤-2相关X蛋白(Bax)、裂解胱天蛋白酶-3(cl-caspase-3)蛋白水平均显著增加(P<0.05);与损伤组比较,阳性对照组及中药低、中药中剂量组、中药高剂量组细胞增殖、Bcl-2、p-PI3K、p-AKT、p-mTOR蛋白表达水平均显著增加(P<0.05),而细胞凋亡率、Bax、cl-caspase-3蛋白水平均显著抑制(P<0.05);且中药组呈剂量依赖。结论:益肾疏肝汤含药血清可能通过激活PI3K/AKT/mTOR通路,抑制卵巢颗粒细胞凋亡。
Objective:To investigate the effect of Yishen Shugan Decoction(YSD)-containing serum on apoptosis of granulosa cells in rats treated with Tripterygium wilfordii glycosides(TWG)and the underlying mechanism.Methods:Ovarian granulosa cells of rats were isolated and treated with different concentration of YSD-containing serum and different concentration of TWG-containing serum for 48 h.Cell survival rate was detected by Cell Counting Kit-8(CCK-8)assay,and the doses of YSD-containing serum and TWG-containing serum were screened.Cells were classified into normal group(normally cultured cells),injury group,low-dose,medium-dose,and high-dose Chinese medicine groups,and positive control group(estradiol-containing serum).Cell proliferation,apoptosis,and expression of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway-related proteins were detected by CCK-8 assay,terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining,and Western blotting.Results:Cell proliferation,and expression of B-cell lymphoma-2(Bcl-2),phosphorylated(p)-PI3K,p-AKT,and p-mTOR were inhibited(P<0.05),and the apoptosis rate and expression of Bcl-2-associated X protein(Bax)and cleaved(cl)-caspase-3 were increased(P<0.05)in the injury group compared with those in the normal group.Cell proliferation,and expression of Bcl-2,p-PI3K,p-AKT,and p-mTOR were increased(P<0.05)and apoptosis rate,and expression of Bax and cl-caspase-3 were inhibited(P<0.05)in positive control group,and low-dose,medium-dose,and high-dose Chinese medicine groups as compared with those in the injury group.The above increase and inhibition were in a dose-dependent manner in the Chinese medicine groups.Conclusion:YSD-containing serum may inhibit the apoptosis of ovarian granulosa cells by activating PI3K/AKT/mTOR pathway.
作者
肖潇
刘艳霞
危一飞
邢玉
滕秀香
XIAO Xiao;LIU Yanxia;WEI Yifei;XING Yu;TENG Xiuxiang(The Second School of Clinical Medicine,Beijing University of Chinese Medicine,Beijing 100078,China;Gynecology of Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University,Beijing 100010,China;Gynecology Department of Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China;Orthopedics Department of Wangjing Hospital,China Academy of Chinese Medical Sciences,Beijing 100102,China)
出处
《世界中医药》
CAS
2022年第22期3148-3153,共6页
World Chinese Medicine
基金
国家自然科学基金项目(81904241)
首都卫生发展科研专项-自主创新项目(首发2022-2-2233)
北京市属医院科研培育计划项目(PZ2018020)。
