Kisspeptin对多囊卵巢综合征大鼠卵巢颗粒细胞凋亡和自噬的影响及机制研究  

Effects of Kisspeptin on apoptosis and autophagy of ovarian granulosa cells in polycystic ovary syndrome rats and the mechanism

作　　者：石明晴[1] 王津[1] 徐小雨 蓝关翠[1] SHI Mingqing;WANG Jin;XU Xiaoyu;LAN Guancui(Department of Obstetrics and Gynecology,Lishui Hospital of Traditional Chinese Medicine,Lishui 323000,China;不详)

机构地区：[1]丽水市中医院妇产科,323000 [2]浙江中医药大学第三临床医学院

出　　处：《浙江医学》2025年第4期342-347,355,I0003,共8页Zhejiang Medical Journal

基　　金：浙江省中医药科学研究基金项目(2021ZB338);浙江省卫生健康科技计划项目(2021RC147)

摘　　要：目的分析Kiss-1基因表达物Kisspeptin(Kp)对多囊卵巢综合征(PCOS)大鼠卵巢颗粒细胞分泌、增殖、凋亡和自噬的影响,探讨Kp对PCOS大鼠卵巢颗粒细胞凋亡和自噬的调控机制。方法采用来曲唑灌胃构建PCOS大鼠模型,分离并培养PCOS大鼠卵巢颗粒细胞,并将细胞分为Kp高、中、低剂量组和模型组,将正常大鼠卵巢颗粒细胞设为正常对照组,共5组。向Kp高、中、低剂量组分别加入浓度1000、100、10 nmol/L的Kp-10培养液。经过24、48、72 h的细胞培养后,采用细胞计数试剂盒8法观察细胞增殖情况,酶联免疫吸附试验检测细胞分泌功能,实时聚合酶链反应检测细胞培养72 h后凋亡因子[肿瘤蛋白53(p53)、细胞色素C(CytC)]、自噬因子[p62蛋白(p62)、Beclin1蛋白(Beclin1)]mRNA的表达水平。结果与正常对照组比较,模型组大鼠卵巢组织形态发生变化,血清雌二醇(E_(2))、卵泡刺激素(FSH)水平降低,血清黄体生成素(LH)、睾酮(T)水平升高,细胞增殖能力下降,细胞分泌孕酮(P)、E_(2)水平下降,p53、CytC、Beclin1 mRNA表达水平升高,p62 mRNA表达水平降低,差异均有统计学意义(均P<0.05)。与模型组比较,卵巢颗粒细胞培养48 h后Kp高、中剂量组细胞增殖能力均上升,培养72 h后Kp高、中、低剂量组细胞增殖能力上升,Kp高、中、低剂量组分泌P水平升高,p53、CytC、Beclin1 mRNA表达水平降低,p62 mRNA表达水平升高,Kp高剂量组分泌E_(2)水平升高,差异均有统计学意义(均P<0.05)。与Kp低剂量组比较,培养48、72 h后Kp高、中剂量组细胞增殖能力升高,Kp高、中剂量组分泌P、E_(2)水平升高,p53、CytC mRNA表达水平降低,p62 mRNA表达水平升高,差异均有统计学意义(均P<0.05)。与Kp中剂量组比较,培养48、72 h后Kp高剂量组细胞增殖能力升高,Kp高剂量组分泌P水平升高,Beclin1 mRNA表达水平降低,差异均有统计学意义(均P<0.05)。结论Kp可促进PCOS大鼠卵巢颗粒细胞增殖及�Objective To analyzed the effects of Kiss-1 gene expression Kisspeptin(Kp)on the secretion,proliferation,apoptosis,and autophagy of ovarian granulosa cells in polycystic ovary syndrome(PCOS)rats,and to explore the regulatory mechanism of Kp on apoptosis and autophagy of ovarian granulosa cells in PCOS rats.Methods A rat model of PCOS was established by gavage with letrozole,and granulosa cells from the ovaries of PCOS rats were isolated and cultured.The granulosa cells from PCOS rats were then divided into KP low-dose group,KP medium-dose group,KP highdose group,and a model group,normal rat ovarian granulosa cells were set as the normal control group,with a total of 5 groups.Different concentrations of Kp-10 culture medium(1000,100,10 nmol/L)were assigned to granular cells of PCOS rat in Kp high-,medium-,and low-dose groups randomly to contrast with the model group of PCOS rat granulosa cells cultured in normal medium.After 24,48,and 72 h of cell culture,cell proliferation were observed by cell counting kit-8,cell secretory function was detected by enzyme-linked immunosorbent assay,and mRNA expressions of apoptosis factors[tumor protein 53(p53),cytochrome C(CytC)]and autophagy(p62,Beclin1)were detected by real-time polymerase chain reaction after 72 h of cell culture.Results Compared with the normal control group,the model group rats showed changes in the morphology of ovarian tissue,decreased levels of serum estradiol(E_(2))and follicle-stimulating hormone(FSH),increased levels of serum luteinizing hormone(LH)and testosterone(T),decreased cell proliferation ability,decreased levels of serum E_(2) and progesterone(P),increased mRNA expression levels of p53,CytC,and Beclin1,and decreased expression of p62 mRNA(all P<0.05).Compared with the model group,the Kp high-and medium-dose groups showed increased cell proliferation ability after culturing the granulosa cells for 48 h,while the cell proliferation ability increased in the Kp high-,medium-,and low-dose groups after culturing for 72 h(all P<0.05);the three group

关 键 词：Kisspeptin/GPR54系统 多囊卵巢综合征 颗粒细胞 凋亡 自噬 

分 类 号：R711.75[医药卫生—妇产科学]