葡萄糖依赖性促胰岛素多肽受体拮抗剂对小鼠多囊卵巢综合征的治疗作用

Therapeutic effects of glucose-dependent insulinotropic polypeptide receptor antagonists on polycystic ovary syndrome in mice

目的探讨葡萄糖依赖性促胰岛素多肽(GIP)受体拮抗剂对多囊卵巢综合征(PCOS)小鼠的治疗作用。方法取30只雌性C57BL/6小鼠,随机分为对照组、模型组、干预组各10只。模型组和干预组采用高脂饲料喂养联合皮下注射脱氢表雄酮(DHEA)建立PCOS模型,对照组给予标准饲料喂养。造模完成后,干预组给予GIP受体拮抗剂腹腔注射,模型组和对照组给予等体积生理盐水,连续28 d。每周称量小鼠体质量并记录。造模第7天起每天行阴道涂片检测动情周期变化。药物干预结束后,取尾静脉血,使用血糖仪检测FPG;眼球取血,ELISA法检测FINS,计算胰岛素抵抗指数(HOMA-IR);ELISA法检测血清睾酮(T)、黄体生成素(LH)、卵泡刺激素(FSH),计算LH/FSH;ELISA法检测血清GIP及瘦素。取血后处死,采用HE染色法观察卵巢组织形态,免疫组化法检测卵巢组织GIP蛋白表达。结果与对照组比较,模型组体质量增加,动情周期紊乱;与模型组比较,干预组体质量降低,动情周期部分恢复。与对照组比较,模型组FPG、FINS及HOMA-IR均升高;与模型组比较,干预组FINS及HOMA-IR均降低(P<0.05或<0.01)。与对照组比较,模型组血清T、LH、FSH升高(P均<0.05);与模型组比较,干预组血清T、LH、FSH降低(P均<0.05);三组LH/FSH比较差异无统计学意义(P均>0.05)。与对照组比较,模型组血清瘦素升高(P均<0.05);与模型组比较,干预组血清瘦素降低(P均<0.05)。与对照组比较,模型组卵巢指数升高(P均<0.05),卵巢组织有明显多囊样改变;与模型组比较,干预组卵巢指数降低(P均<0.05),卵巢组织多囊样改变有所改善。与对照组比较,模型组血清及卵巢GIP升高;与模型组比较,干预组血清及卵巢GIP降低(P均<0.05)。结论GIP受体拮抗剂可减轻PCOS小鼠体质量,恢复动情周期,改善胰岛素抵抗,下调血清睾酮,一定程度上恢复PCOS小鼠卵巢多囊样改变。

Objective To investigate the therapeutic effects of glucose-dependent insulinotropic polypeptide(GIP)receptor antagonists on polycystic ovary syndrome(PCOS)in mice.Methods Thirty female C57BL/6 mice were taken and randomly divided into the control,model and intervention groups,with 10 mice in each.Mice in the model and intervention groups were fed with high-fat diet combined with subcutaneous injection of dehydroepiandrosterone(DHEA)to establish the PCOS models,and mice in the control group were fed with standard diet.After modeling,the intervention group was given intraperitoneal injection of GIP receptor antagonist,and the model and control groups were given equal volume of normal saline for 28 d.The body mass of the mice was weighed and recorded weekly.Vaginal smears were taken daily from the 7th day of modeling to detect changes in the estrous cycle.At the end of the pharmacological intervention,blood was taken from the tail vein and FPG was measured using a glucometer;blood was taken from the eyeballs,and FINS was measured by ELISA to calculate the insulin resistance index(HOMA-IR);serum testosterone(T),luteinizing hormone(LH),and follicle-stimulating hormone(FSH)were measured by ELISA to calculate LH/FSH;and serum GIP and leptin were measured by ELISA.After the blood was taken,mice were executed,the morphology of ovarian tissues was observed by HE staining,and GIP protein expression in ovarian tissues was detected by immunohistochemistry.Results Compared with the control group,the model group had increased body mass and disordered estrous cycle;compared with the model group,the intervention group had decreased body mass and partially restored estrous cycle.Compared with the control group,FPG,FINS and HOMA-IR increased in the model group;compared with the model group,FINS and HOMAIR decreased in the intervention group(P<0.05 or P<0.01).Compared with the control group,serum T,LH and FSH were elevated in the model group(all P<0.05);compared with the model group,serum T,LH and FSH were reduced in the intervention group(all P<0.05);no significant difference was found in the LH/FSH among the three groups(P>0.05).Compared with the control group,serum leptin was elevated in the model group(P<0.05);compared with the model group,serum leptin decreased in the intervention group(P<0.05).Compared with the control group,ovarian index was elevated in the model group(P<0.05),and there were obvious polycystic changes in the ovarian tissues;compared with the model group,ovarian index was reduced in the intervention group(P<0.05),and polycystic changes in the ovarian tissues were improved.Compared with the control group,serum and ovarian GIP were elevated in the model group;compared with the model group,serum and ovarian GIP were reduced in the intervention group(both P<0.05).Conclusion GIP receptor antagonists reduced body mass,restored the estrous cycle,improved insulin resistance,down-regulated serum testosterone,and restored ovarian polycystic changes to some extent in PCOS mice.