Chemerin对卵巢颗粒细胞的影响及对PCOS发病机制的研究

Effect of Chemerin on ovarian granulosa cells and pathogenesis of polycystic ovary syndrome

摘要:

目的：探究Chemerin在多囊卵巢综合征（polycystic ovarian syndrome，PCOS）发病机制中的作用。方法：体外培养人卵巢颗粒细胞（COV434），分别下调及过表达COV434细胞中的Chemerin基因，采用ELISA测定培养液炎性因子白介素-6（inter－leukin-6，IL-6）、白介素-8（interleukin-8，IL-8）、白介素-10（interleukin-10，IL-10）及肿瘤坏死因子-α（tumor necrosis factor，TNF-α）浓度，EdU方法测定各组细胞增殖能力，流式细胞法测定细胞凋亡。结果：下调组（LV3-shChemerin-1、LV3-shChemerin-2）与阴性对照组（LV3-NC）相比，培养液中IL-6、IL-8、TNF-α浓度均明显降低，IL-10浓度明显升高，差异均有统计学意义（P IL-6=0.006，PIL-8=0.001，PTNF-α=0.012，PIL-10=0.002），颗粒细胞COV434增殖能力明显降低（P=0.004），凋亡明显增加（P=0.000）。过表达组（LV5-Chemerin）与阴性对照组（LV5-NC）相比，培养液中IL-6、IL-8、TNF-α明显升高，IL-10明显降低，差异均有统计学意义（PIL-6=0.044，PIL-8=0.008，PTNF-α=0.015，PIL-10=0.004），颗粒细胞COV434增殖能力明显提高（P=0.013），凋亡明显降低（P=0.029）。结论：Chemerin可能通过促进卵巢颗粒细胞炎症反应，并诱导颗粒细胞增殖抑制凋亡参与PCOS的发生发展。

Abstract:

Objective：To investigate the role of Chemerin in the pathogenesis of polycystic ovarian syndrome（PCOS）. Methods：Hu－man ovarian granulosa cells（COV434） were cultured in vitro. Down-regulation and overexpression of Chemerin were performed in the COV434 cells. Enzyme-linked immunosorbent assay was used to determine the concentrations of inflammatory factors such as interleukin-6（IL-6），interleukin-8（IL-8），and interleukin-10（IL-10），and tumor necrosis factor-α（TNF-α） in the culture medium. The EdU method and flow cytometry were used to measure cell proliferation ability and apoptosis，respectively. Results：Compared with the negative control group（LV3-NC），the down-regulation groups（LV3-shChemerin-1，LV3-shChemerin-2） had significantly lower concentrations of IL-6，IL-8，and TNF-α and a significantly higher concentration of IL-10 in the culture medium（PIL-6=0.006，PIL-8=0.001，PTNF-α=0.012，PIL-10=0.002），significantly lower proliferative capacity of COV434 cells（P=0.004），and signifi－cantly enhanced apoptosis（P=0.000）. Compared with the negative control group（LV5-NC），the overexpression group（LV5-Chemerin） had significantly higher concentrations of IL-6，IL-8，and TNF-α and a significantly lower concentration of IL-10 in the culture medium（PIL-6=0.044，PIL-8=0.008，PTNF-α=0.015，PIL-10=0.004），significantly higher proliferative capacity of COV434 cells（P=0.013），and significantly reduced apoptosis（P=0.030）. Conclusion：Chemerin plays a role in the development and progression of PCOS by promoting inflammatory response，inducing proliferation，and inhibiting apoptosis of ovarian granulosa cells.