血小板反应蛋白-1在PCOS大鼠卵巢组织的表达及对卵巢微血管内皮细胞迁移的影响  

Expression of thrombospondin-1 in ovarian tissue of PCOS rats and its effect on migration of ovarian microvascular endothelial cells

作　　者：严谨 王超[1] 王瑞静 刘羽[1] 张琰惠 王雅新 张婷[1] 刘梅梅[1] Yan Jin;Wang Chao;Wang Ruijing;Liu Yu;Zhang Yanhui;Wang Yaxin;Zhang Ting;Liu Meimei(Department of Obstetrics and Gynecology,the Second Affiliated Hospital of Harbin Medical University,Harbin Heilongjiang 150081,P.R.China)

机构地区：[1]哈尔滨医科大学附属第二医院妇产科,黑龙江哈尔滨150081

出　　处：《中国计划生育和妇产科》2023年第9期53-58,F0003,共7页Chinese Journal of Family Planning & Gynecotokology

基　　金：吴阶平基金会临床科研专项资助基金(项目编号:320.6750.18542)。

摘　　要：目的探讨来曲唑诱导的多囊卵巢综合征(polycystic ovary syndrome,PCOS)模型大鼠卵巢组织中血小板反应蛋白-1(TSP-1)的表达和意义及在体外对大鼠原代卵巢微血管内皮细胞迁移的影响。方法将40只雌性SD大鼠随机分为对照组10只,模型组30只。模型组予来曲唑(1 mg/kg·d)连续灌胃21 d建立PCOS大鼠模型,对照组予等剂量的羧甲基纤维素钠灌胃。造模成功后,模型组随机筛选10只为治疗组。治疗组予TSP-1溶液(2 mg/kg·d)腹腔注射,连续21 d。检测各组血清性激素水平,观察大鼠卵巢形态学及组织学变化,蛋白质免疫印迹方法(Western Blot)检测卵巢组织中TSP-1的表达,提取模型组大鼠原代卵巢微血管内皮细胞并进行体外培养,划痕实验检测TSP-1对模型组大鼠卵巢微血管内皮细胞迁移的影响。结果模型组与对照组比较,血清睾酮(T)、黄体生成素(LH)水平及LH/FSH比值升高(P<0.05),卵巢组织中TSP-1表达水平降低(P<0.05)。模型组大鼠卵巢呈多囊改变。治疗组与模型组比较,血清T、LH水平及LH/FSH比值降低,卵巢组织TSP-1蛋白表达水平增加,卵巢形态学改善。在体外加入TSP-1后,与未加入TSP-1的卵巢微血管内皮细胞相比,细胞迁移率明显降低(P<0.05),且随TSP-1浓度升高,迁移率明显降低(P<0.05);与加入TSP-1(100 ng/mL)细胞相比,加入TSP-1(100 ng/mL)+P38 MAPK通路抑制剂(SB203580)(5μmol/L)的细胞迁移率明显增加(P<0.05)。结论在来曲唑诱导的PCOS模型大鼠卵巢组织中TSP-1表达降低,腹腔注射外源性TSP-1可在一定程度上改善PCOS大鼠的血清性激素水平和卵巢异常组织形态。在体外,TSP-1可以抑制PCOS大鼠卵巢微血管内皮细胞迁移能力,且呈浓度依赖性,P38 MAPK通路可能在其中发挥作用。Objective To investigate the expression and significance of thrombospondin-1(TSP-1)in ovarian tissue of letrozole-induced polycystic ovary syndrome(PCOS)model rats and the effect of TSP-1 on the migration of primary ovarian microvascular endothelial cells in vitro of rats.Methods Forty female SD rats were randomly divided into control group(n=10)and model group(n=30).The model group was given letrozole(1 mg/kg·d)by gavage for 21 days to establish the PCOS rat model,and the control group was given the same dose of sodium carboxymethyl cellulose by the using of gavage.After successful modeling,10 rats in the model group were randomly selected as the treatment group.The treatment group received intraperitoneal injection of TSP-1 solution(2 mg/kg·d)for 21 consecutive days.The levels of serum sex hormones in each group were detected,the morphological and histological changes of the ovaries of the rats were observed,the expression of TSP-1 in the ovarian tissues was detected by Western Blot,and the primary ovarian microvascular endothelial cells of the rats in the model group were extracted and analyzed in vitro.The effect of TSP-1 on the migration of ovarian microvascular endothelial cells in the model group was detected by scratch test.Results Compared with the control group,the levels of serum testosterone(T),luteinizing hormone(LH)and LH/FSH ratio in the model group increased(P<0.05),and the expression level of TSP-1 in ovarian tissue decreased(P<0.05).The ovaries in the model group showed polycystic changes.Compared with the model group,the serum T,LH levels and LH/FSH ratio were decreased in the treatment group,the expression level of TSP-1 protein in ovarian tissue was increased,and the ovarian morphology was improved.After adding TSP-1 in vitro,compared with the ovarian microvascular endothelial cells without TSP-1,the cell migration rate was significantly decreased(P<0.05),and with the increase of TSP-1 concentration,the migration rates was significantly decreased(P<0.05).Compared with the addition of TSP-1(

关 键 词：多囊卵巢综合征 血小板反应蛋白-1 内皮细胞迁移 大鼠 

分 类 号：R711.75[医药卫生—妇产科学]