Kiss-1/GPR54系统在PCOS模型大鼠卵巢颗粒细胞中的表达及对卵泡发育障碍的作用机制  

Expression of Kiss-1/GPR54 system in ovarian granulosa cells of PCOS rats and its mechanism in follicular developmental disorders

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,Zhejiang,China;The Third Clinical Medical College of Zhejiang Chinese Medicine University,Hangzhou 310053,Zhejiang,China)

机构地区：[1]浙江省丽水市中医院妇产科,丽水323000 [2]浙江中医药大学第三临床医学院,杭州310053

出　　处：《浙江中西医结合杂志》2024年第11期994-998,1005,共6页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省中医药科技计划项目(No.2021ZB338);浙江省卫生健康科技计划项目(No.2021RC147)。

摘　　要：目的初步探讨Kiss-1/GPR54系统在多囊卵巢综合征(PCOS)模型大鼠卵巢颗粒细胞中的表达,并分析其与PCOS之间的关联。方法筛选出14只动情规律的雌性SD大鼠,按照随机数字表法分为正常对照组和模型组,每组7只。其中模型组大鼠予来曲唑-羧甲基纤维素混悬液0.1mg/(kg·d)连续灌胃21 d以构建PCOS大鼠模型。收集大鼠血清、卵巢组织、卵巢颗粒细胞。然后计算大鼠卵巢指数,对卵巢组织进行苏木精-伊红染色法,采用酶联免疫吸附测定法检测血清激素水平,免疫荧光染色检测颗粒细胞中亲吻素(Kp)、G蛋白偶联受体54(GPR54)荧光强度,实时定量反转录聚合酶链式反应检测颗粒细胞中Kiss-1、GPR54基因表达。结果与正常对照组比较,模型组大鼠卵巢囊状扩张卵泡及闭锁卵泡增加,卵泡颗粒细胞层变薄;卵巢指数增大[(1.22±0.10)mg/g比(0.82±0.13)mg/g,P<0.05];血清黄体生成素[(66.32±3.98)IU/L比(11.87±5.54)IU/L]、睾酮[(33.72±3.57)ng/mL比(8.40±2.94)ng/mL]、Kp水平[(1506.79±154.82)pg/mL比(289.57±89.20)pg/mL]均升高(P<0.05);卵巢颗粒细胞中Kp荧光强度[(1601852.67±378567.82)比(3685156.00±359825.63)]、GPR54荧光强度[(1298372.25±297701.61)比(2961456.58±309119.01)]及Kiss-1基因表达[(0.10±0.03)比(1.11±0.14)]、GPR54基因表达[(0.10±0.05)比(1.00±0.13)]均降低(P<0.05)。结论卵巢表达的Kiss-1/GPR54系统可能通过调节颗粒细胞从而影响PCOS大鼠卵泡发育。Objective To preliminarily investigate the expression of Kiss-1/GPR54 system in ovarian granulosa cells of a polycystic ovary syndrome(PCOS)rat model and analyze its association with PCOS.Methods Fourteen female SD rats with regular estrous cycles were randomly divided into two groups using random number table method:the normal control group and the PCOS model group,with seven rats per group.The model group received 0.1 mg/(kg·d)letrozole carboxymethyl cellulose suspension by gavage for 21 consecutive days to induce PCOS.The serum,ovarian tissue,and ovarian granulosa cells were collected from all rats.The ovarian index was calculated,and ovarian tissues were stained with hematoxylin-eosin.An enzyme-linked immunosorbent assay(ELISA)was used to detect serum hormone levels,and immunofluorescence staining was used to detect the fluorescence intensity of kisspeptin(Kp)and G protein-coupled receptor 54(GPR54)in granulosa cells.Real-time quantitative reverse transcription polymerase chain reaction(RT-PCR)was used to detect Kiss-1 and GPR54 gene expression in granulosa cells.Results Compared to the normal control group,the model group presented significantly increased numbers of cystic and atretic follicles,thinner granulosa cell layer,and increased ovarian index[(1.22±0.10)mg/g vs.(0.82±0.13)mg/g,P<0.05].The serum levels of luteinizing hormone[(66.32±3.98)IU/L vs.(11.87±5.54)IU/L],testosterone[(33.72±3.57)ng/mL vs.(8.40±2.94)ng/mL],and Kp[(1506.79±154.82)pg/mL vs.(289.57±89.20)pg/mL]were all increased(P<0.05).The fluorescence intensity of Kp[(1601852.67±378567.82)vs.(3685156.00±359825.63)]and GPR54[(1298372.25±297701.61)vs.(2961456.58±309119.01)],and the gene expression of Kiss-1[(0.10依0.03)vs.(1.11依0.14)]and GPR54[(0.10依0.05)vs.(1.00依0.13)]in ovarian granulose cells were significantly decreased(P<0.05).Conclusion The Kiss-1/GPR54 system in the ovaries may affect follicular development in PCOS rats by regulating granulosa cells.

关 键 词：大鼠 多囊卵巢综合征 颗粒细胞 卵泡发育 亲吻素 G蛋白偶联受体54 

分 类 号：R711.75[医药卫生—妇产科学]