黄芩素对雷公藤甲素诱导的大鼠卵巢颗粒细胞凋亡的保护作用  

Protective Effects of Baicalein on Triptolide-induced Ovarian Granulosa Cells Apoptosis in Rats

作　　者：陶旻枫[1] 于敬龙[2] 杨维[1] 

机构地区：[1]首都医科大学附属北京妇产医院北京妇幼保健院,北京100026 [2]北京中医药大学东直门医院,北京100700

出　　处：《上海中医药大学学报》2017年第3期85-89,共5页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：中国疾病预防控制中心科研基金项目(2015FY004)

摘　　要：目的:探讨黄芩素对雷公藤甲素诱导的大鼠卵巢颗粒细胞凋亡的保护作用。方法:收集大鼠卵巢颗粒细胞,经原代培养24 h后,用雷公藤甲素损伤大鼠卵巢颗粒细胞;干预组添加终浓度分别为50、100、200μmol/L的黄芩素,对照组添加相同体积的二甲基亚砜(DMSO)溶液,继续培养24 h(或以上)。采用MTT法测定卵巢颗粒细胞活力,流式细胞术Annexin V/PI双染色检测细胞凋亡,Western blot法检测与细胞凋亡相关蛋白的表达情况;测定各组卵巢颗粒细胞超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性及丙二醛(MDA)含量,评价抗氧化能力。结果:雷公藤甲素可显著降低大鼠卵巢颗粒细胞活力,诱导细胞凋亡,并使SOD和GSH-Px活性降低,MDA含量增加;雷公藤甲素处理细胞的同时添加黄芩素干预后,卵巢颗粒细胞相对活力增加,细胞凋亡减弱,胞内SOD和GSH-Px活性升高,MDA含量降低。Western blot结果显示,经过雷公藤甲素处理,卵巢颗粒细胞中促凋亡蛋白Bax表达量显著上升,抗凋亡蛋白Bcl-2表达量明显降低;黄芩素干预后可以显著下调Bax表达量,上调Bcl-2表达量。结论:黄芩素可明显改变卵巢颗粒细胞的抗凋亡及抗氧化能力,对雷公藤甲素诱导的细胞凋亡起保护作用。Objective： To investigate the protective effects of baicalein on triptolide-induced ovarian granulosa cells apoptosis in rats. Methods ： The ovarian granulosa cells of rats were collected and primary cultured for 24 hours. Triptolide was added to damage the ovarian granulosa ceils. The intervention groups were treated with baicalein at the final concentrations of 50,100,200 μmol/L,the control group was treated with dimethyl sulfoxide （DMSO） solution with the same volume. Then the cells were cultured for 24 hours or more. The viability of ovarian granulosa cells was measured by MTT assay, the cell apoptosis was detected by flow cytometry Annexin V/PI double staining. The expressions of apoptosis related proteins were determined by Western blot. The activities of supemxide dismutase （SOD） and glutathione pemxidase （GSH-Px） and the content of malondialchehyche （MDA） were detected to evaluate the antioxidant effects. Results： Triptolide could significantly decrease the viability of ovarian granulosa cells, induce the cell apoptosis, reduce the activities of SOD and GSH-Px, and increase the MDA content. After the intervention of baicalein on triptolide-treated cells, the relative viability of ovarian granulosa cells was increased, the cells apoptosis was reduced, the intracellular activities of SOD and GSH-Px were increased and the MDA content was decreased. The results of Western Blot showed that the expression level of pro-apoptotic protein Bax was significantly increased in the ovarian granulosa ceils, and the expression level of anti-apoptotic protein Bcl-2 was significantly decreased after triptolide treatment. While after baicalein intervention, the Bax expression was significantly down-regulated and the Bcl- 2 expression was significantly up-regulated. Conclusion： Baicalein can significantly improve the anti-apoptotic and antioxidant effects of ovarian granulosa cells, which plays the protective role in the cell apoptosis induced by triptolide.

关 键 词：黄芩素 雷公藤甲素 卵巢颗粒细胞 氧化应激 凋亡 

分 类 号：R285.5[医药卫生—中药学]