氯吡格雷抑制PDGF的表达改善大鼠宫腔粘连

Inhibition of PDGF Expression by Clopidogrel Improves Intrauterine Adhesion in Rats

目的:本文拟研究氯吡格雷对改善大鼠宫腔粘连的作用及其可能的机制,为临床治疗提供理论依据。方法:30只雌性SD大鼠随机分为3组:正常对照组(NC)(n=10)、宫腔粘连模型组(IUA)(n=10)和氯吡格雷处理组(IUA+Clo)(n=10)。采用机械损伤法建立IUA模型。建模7天后,氯吡格雷处理组大鼠通过灌胃给予氯吡格雷治疗,60 mg/kg/d;模型组大鼠通过灌胃给予等剂量生理盐水,持续7天。苏木精-伊红(HE)染色法检测大鼠子宫组织病理学变化,计算子宫内膜腺体数量及测量子宫内膜厚度。Masson染色法检测大鼠子宫组织纤维化变化。酶联免疫吸附实验(ELISA)检测大鼠血清中炎性因子TNF-α和IL-6表达水平。免疫组化法(IHC)检测大鼠子宫组织PDGF、collagenⅠ和α-SMA表达水平。实时定量聚合酶链式反应(qRT-PCR)检测子宫组织PDGF、collagenⅠ、α-SMA m RNA表达水平。蛋白免疫印迹法(Western blot)检测子宫组织PDGF、collagenⅠ、α-SMA蛋白表达水平。结果:与正常对照组相比,宫腔粘连模型组大鼠子宫内膜腺体数目显著减少(P<0.05),子宫内膜变薄、纤维化程度明显增加(P<0.05)。与宫腔粘连模型组相比,氯吡格雷处理组大鼠子宫内膜腺体数目显著增加(P<0.05),子宫内膜显著增厚(P<0.05),血清炎性因子TNF-α、IL-6水平明显降低(P<0.05),子宫组织PDGF、collagenⅠ、α-SMA表达均显著降低(P<0.05)。结论:氯吡格雷能够抑制大鼠体内血小板的聚集活化,抑制炎症水平,降低PDGF表达,抑制大鼠子宫内膜纤维化的发生及进展。氯吡格雷可能成为临床治疗宫腔粘连的药物之一。

Objective:Intrauterine adhesion(IUA)is a common disease in women of childbearing age and one of the common causes of female infertility.This study proposes whether clopidogrel can improve intrauterine adhesion in rats and its possible mechanism.Providing a theoretical basis for clinical treatment.Methods:Thirty female Sprague-Dawley rats were randomly divided into 3 groups:normal control group(NC)(n=10,gavage with normal saline),intrauterine adhesion model group(IUA)(n=10,gavage with normal saline)and clopidogrel treatment group(IUA+Clo)(n=10,gavage with clopidogrel).The mechanical damage method was used to establish the IUA model,after 7 days of modeling,the rats in the clopidogrel treatment group were treated with clopidogrel by gavage,60 mg/kg/d.Rats in the model group were given the same dose of normal saline by gavage for 7 days.Hematoxylin-eosin(HE)staining was used to detect the pathological changes of the uterus,calculate the number of endometrial glands and measure the thickness of the endometrium.Masson staining was used to detect the fibrosis of rat uterus.The expression levels of TNF-αand IL-6 in serum of rats were detected by enzyme-linked immunosorbent assay(ELISA).The expression levels of PDGF,collagenⅠandα-SMA in rat uterine tissues were detected by immunohistochemistry(IHC).Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression levels of PDGF,collagenⅠandα-SMA in uterine tissue.The protein expression levels of PDGF,collagenⅠandα-SMA in uterine tissue were detected by Western blot.Results:Compared with that in the normal control group,the number of endometrial glands in the intrauterine adhesion model group reduced significantly(P<0.05).The degree of endometrial thinning and fibrosis increased significantly(P<0.05).Compared with that in the intrauterine adhesion model group,the number of endometrial glands in the clopidogrel treatment groups increased significantly(P<0.05).endometrium exhibited a significant increase(P<0.05),the levels of serum inflammatory factors TNF-αand IL-6 decreased significantly(P<0.05).The expressions of PDGF,collagenⅠandα-SMA in uterine tissue decreased significantly(P<0.05).Conclusion:Clopidogrel can inhibit the aggregation and activation of platelets,inhibit the level of inflammation,reduce the expression of PDGF and inhibit the occurrence and progression of endometrial fibrosis in rats,Clopidogrel may be one of the drugs for the treatment of intrauterine adhesions.