人参皂苷Rh_(2)在卵巢颗粒细胞炎性反应中的作用及作用机制网络药理学与分子生物学研究  

Role and Mechanism of Ginsenoside Rh_(2) in the Inflammatory Response of Ovarian Granulosa Cells Based on Network Pharmacology and Molecular Biology

作　　者：王文芳[1] 海鑫 WANG Wenfang;HAI Xin(The First Affiliated Hospital of Harbin Medical University,Harbin,Heilongjiang,China 150086)

机构地区：[1]哈尔滨医科大学附属第一医院,黑龙江哈尔滨150086

出　　处：《中国药业》2023年第1期46-51,共6页China Pharmaceuticals

基　　金：黑龙江省自然科学基金优秀青年项目[YQ2019H016]。

摘　　要：目的探讨西洋参中人参皂苷Rh(2简称Rh_(2))在脂多糖(LPS)致人卵巢颗粒细胞瘤细胞系KGN细胞炎性反应中的作用及作用机制。方法通过网络药理学方法筛选西洋参治疗多囊卵巢综合征(PCOS)的潜在活性成分及靶点。以200 ng/mL LPS作用KGN细胞6 h诱导炎性反应,采用酶联免疫吸附(ELISA)法测定KGN细胞中炎性因子白细胞介素1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的水平,采用2',7'-二氯双氢荧光素双乙酸酯(DCFH-DA)染色法测定活性氧(ROS)的水平;40µmol/L Rh_(2)作用KGN细胞24 h后,采用免疫印迹(Western blot)法测定哺乳动物雷帕霉素靶蛋白(mTOR)在LPS诱导的KGN细胞中的表达水平。结果网络药理学分析结果显示,共筛选出西洋参的主要化学成分11个、潜在靶点144个、治疗PCOS的靶点13个,其中Rh_(2)与下游靶基因mTOR是西洋参抗PCOS的关键活性成分及潜在作用靶点。分子生物学研究结果显示,Rh_(2)能抑制LPS导致的KGN细胞中ROS和mTOR表达水平的升高,下调IL-1β和TNF-α的表达;低表达(50 nmol/L)mTOR能促进Rh_(2)缓解LPS导致的KGN细胞炎性反应,过表达(3µg)mTOR能逆转此现象。结论Rh_(2)通过调控mTOR的表达水平参与LPS导致的卵巢颗粒细胞炎性反应。Objective To investigate the role and mechanism of ginsenoside Rh_(2)(hereinafter referred to as Rh_(2))in the inflammatory response of human ovarian granulosa tumor cell line KGN induced by lipopolysaccharide(LPS).Methods The potential active components and targets of Panacis Quinquefolii Radix in the treatment of polycystic ovary syndrome(PCOS)were screened based on the network pharmacology.The KGN cells were treated with 200 ng/mL LPS for 6 h to induce the inflammatory response.The levels of inflammatory factors including interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the KGN cells were detected by the enzyme-linked immunosorbent assay(ELISA)method.The reactive oxygen species(ROS)level in the KGN cells was detected by the 2',7'-dichlorodihydrofluorescein-diacetate(DCFH-DA)staining method.After the KGN cells were treated with 40µmol/L Rh_(2) for 24 h,the expression level of mammalian target of rapamycin(mTOR)in the KGN cells induced by LPS was detected by the Western blot.Results The results of network pharmacology showed that 11 main chemical components,144 potential targets and 13 targets in the treatment of PCOS in Panacis Quinquefolii Radix were screened out.Among them,Rh_(2) and the downstream target gene mTOR were the key active component and potential target of Panacis Quinquefolii Radix in the treatment of PCOS.The results of molecular biology showed that Rh_(2) could inhibit the high expression of ROS and mTOR and downregulate the expression of IL-1βand TNF-αin the KGN cells induced by LPS.Low expression of mTOR(50 nmol/L)could promote Rh_(2) to alleviate the inflammatory response of KGN cells induced by LPS,while overexpression of mTOR(3µg)could reverse this phenomenon.Conclusion Rh_(2) participates in the inflammatory response of ovarian granulosa cells induced by LPS by regulating the expression of mTOR.

关 键 词：多囊卵巢综合征 卵巢颗粒细胞 人参皂苷Rh_(2) 西洋参 哺乳动物雷帕霉素靶蛋白 炎性反应 网络药理学 分子生物学 

分 类 号：R932[医药卫生—生药学] R285.5[医药卫生—药学]