PI3k/Akt通路参与二仙汤抑制顺铂所致卵巢颗粒细胞凋亡的作用  

Effects of Er-Xian Decoction on Cisplatin Induced Ovarian Granulosa Cells Apoptosis Through PI3k/Akt Pathway

作　　者：杨蕾 陶仕英 王继峰 牛建昭 赵丕文 孙丽萍 王燕霞 武洪波 蔡欣悦 杨阳 

机构地区：[1]北京中医约大学基础医学院,北京100029

出　　处：《世界科学技术-中医药现代化》2016年第8期1362-1367,共6页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：教育部国家外专局高等学校学科创新引智基地项目(B07007)负责人:牛建昭;国家自然科学基金委资助项目(81273887)基于PI3k/Akt信号通路探讨二仙汤治疗化疗损伤性卵巢早衰的作用机制,负责人:牛建昭;国家自然科学基金委资助项目(30772849):报告基因技术研究二仙汤的雌激素作用机制和配伍规律,负责人:王继峰

摘　　要：目的:观察顺铂干预大鼠卵巢颗粒细胞凋亡情况,从而探讨二仙汤含药血清经PI3k/Akt通路抑制卵巢颗粒细胞凋亡的作用机制。方法:SD雌性大鼠30只,随机分为3组,分别为生理盐水组、补佳乐组、二仙汤组。心脏取血,制备药物血清。SD大鼠卵巢颗粒细胞,经培养贴壁后,用顺铂损伤大鼠原代卵巢颗粒细胞。经各组药理血清作用后,应用Annexin V/PI双染流式细胞仪检测细胞凋亡,并对各实验组大鼠卵巢颗粒细胞的凋亡情况进行分析。应用免疫组化法,测定各分组PI3k、Akt、p-Akt蛋白的表达情况,并进行图像分析。结果:流式细胞凋亡结果显示,二仙汤可以降低顺铂损伤后的大鼠卵巢颗粒细胞的凋亡率。免疫组化结果显示,PI3k、p-Akt、Akt在细胞质内均有表达,可见棕黄色阳性物质;而二仙汤可以上调顺铂干预后大鼠卵巢颗粒细胞的PI3k、p-Akt、Akt的表达。结论:二仙汤含药血清上调了PI3k、p-Akt、Akt的表达,从而抑制了卵巢颗粒细胞凋亡,该过程可能通过调节PI3k/Akt通路实现,进而有效地保护卵巢功能。This study aimed to investigate the effects of cisplatin on rats＇ ovarian granulosa （OG） cells apoptosis, and to explore the mechanism of Er-xian Decoction （EXD） behind the anti-apoptotic role of its pharmacological serum in OG cells by PI3k/Akt pathway.30 female SD rats were randomly divided into three groups, the normal saline group, the progynova group and the EXD group. Pharmacological serum was prepared after heart puncture. Prior to cisplatin stimulation, SD rat OG cells were cultivated withadherence method.When administered by pharmacological serum of EXD, AnnexinV/PI double staining flow cytometry was adopted to detect cell apoptotic situations of OG cells of all groups, and immunohistochemical technology to quantify the protein expressions of PI3k, Akt and p-Akt combined with and the image analysis.h was found that the apoptosis rate, stimulated by cisplatin, was reducedafler EXD administration according to analysis of flow cytometry. Immunohistochemical outcomes showed that PI3k, p-Akt and Akt were expressed in the cytoplasm featuring brown positive substances. EXD enhanced the expressions of PI3k, p-Akt and Akt of cisplatin-stimulated OG cells. In conclusion, EXD prohibited the apoptosis of OG cells through upregulating the expressions of PI3k, p-Akt and Akt, by which affected the PI3k/Akt pathway further and effectively protectedthe ovarian function.

关 键 词：二仙汤 卵巢颗粒细胞 PI3K/AKT通路 细胞凋亡 

分 类 号：R285[医药卫生—中药学]