富血小板血浆治疗小鼠薄型子宫内膜的效果及机制研究  

The effect and mechanism of platelet-rich plasma therapy on thin endometrium in mice

作　　者：王晓寒 刘玲[3,4] 牟善茂[2] 张敏 孙朝霞 郑洋洋 郝翠芳 WANG Xiaohan;LIU Ling;MOU Shanmao;ZHANG Min;SUN Zhaoxia;ZHENG Yangyang;HAO Cuifang(Department of Obstetrics and Gynecology,Rizhao Hospital of Traditional Chinese Medicine,Shandong Rizhao 276800,China;Department of Encephalopathy,Rizhao Hospital of Traditional Chinese Medicine,Shandong Rizhao 276800,China;Centre for Reproductive Medicine,Affiliated Weihai Second Municipal Hospital of Qingdao University,Shandong Qingdao 264200,China;Shandong University,Jinan 250100;Reproductive Health Center,Women and Children’s Hospital,Qingdao University,Shandong Qingdao 266011,China)

机构地区：[1]日照市中医医院妇产科,山东日照276800 [2]日照市中医医院脑病科,山东日照276800 [3]青岛大学附属威海市立第二医院生殖医学中心,山东威海264200 [4]山东大学,济南250100 [5]青岛大学附属妇女儿童医院生殖中心,山东青岛266011

出　　处：《中国妇产科临床杂志》2023年第4期401-405,共5页Chinese Journal of Clinical Obstetrics and Gynecology

基　　金：山东省医药卫生科技发展计划项目(202005031192);日照市优秀青年科学基金项目(RZ2021ZR24)。

摘　　要：目的探讨宫腔灌注富血小板血浆(platelet-rich plasma,PRP)治疗薄型子宫内膜的效果及机制。方法取18只动情周期规律的成熟未交配雌性小鼠,随机数字法分为正常对照组(Control组)、模型对照组(Model组)和PRP治疗组(PRP组),每组均6只。Model组及PRP组小鼠行宫腔注射95%乙醇制作薄型子宫内膜模型后,PRP组宫腔灌注PRP 100μl,Model组宫腔灌注生理盐水100μl,每隔3天灌注一次,共3次,Control组不行任何处理。3个动情周期后取材。通过HE染色观察子宫内膜厚度、面积及微血管改变;免疫组化、ELISA和Western-blot检测子宫内膜中角蛋白(cytokeratin,CK9)、波形蛋白(vimentin,VIM)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、白血病抑制因子(leukemia inhibitory factor,LIF)和白介素-6(Interleukin-6,IL-6)的表达;并通过免疫荧光双染法检测小鼠子宫内膜间充质干细胞(endometrial mesenchymal stem cells,EnMSCs)的阳性表面标志物:CD90和CD105,并观察EnMSCs的在体变化情况。结果PRP组小鼠子宫内膜面积[(713164.67±81618.56)μm^(2)]明显高于Model组[(434001.67±28595.37)μm^(2),P<0.05];PRP组小鼠的子宫内膜容受性指标:CK9、VIM、VEGF、LIF蛋白的表达量均明显高于Model组(P<0.05);而炎性指标IL-6[(25.75±2.35)pg/mL]明显低于Model组[(35.33±1.96)pg/mL,P<0.05]。PRP组小鼠在体EnMSCs的数量[(0.13±0.04)%]低于Control组[(0.22±0.06)%,P<0.05],与Model组比较,差异无统计学意义(P>0.05)。结论PRP宫腔灌注薄型子宫内膜,可促进受损子宫内膜增殖修复,改善子宫内膜容受性,减轻子宫内膜炎症反应。Objective To investigate the effect and mechanism of platelet-rich plasma(PRP)treating thin endometrium.Methods Eighteen mature unmated female mice with regular estrous cycle were randomly divided into Control group,Model group and PRP group,with 6 mice in each group.Model group and PRP group were perfused with 95%ethanol into uterine cavity to make thin endometrial model.After operation,PRP group was perfused with PRP 100μl and Model group was perfused with normal saline 100μl every 3 days for 3 times.Control group had no management.Samples were obtained after 3 estrous cycles.The endometrial thickness,area and microvascular changes were observed by HE staining.The expressions of cytokeratin(CK9),vimentin(VIM),vascular endothelial growth factor(VEGF),leukemia inhibitory factor(LIF)and interleukin-6(IL-6)were detected by immunohistochemistry,ELISA and Western-blot.Immunofluorescence double staining was used to detect the positive markers of endometrial mesenchymal stem cells(EnMSCs)in mice uterus including CD90 and CD105 to observe the EnMSCs changes situation.Results The endometrial area of PRP group(713164.67±81618.56)μm^(2)was significantly higher than that of Model group[(434001.67±28595.37)μm^(2),P<0.05].The protein expression of endometrial tolerance markers CK9,VIM,VEGF,and LIF were significantly higher in PRP group compared with Model group(P<0.05),but with lower level of IL-6(25.75±2.35 pg/ml)than Model group[(35.33±1.96 pg/ml),P<0.05].Although the number of EnMSCs(0.13±0.04%)in vivo in PRP group was lower than that in Control group[(0.22±0.06%),P<0.05],there was not statistically different from the Model group(P>0.05).Conclusions PRP intrauterine perfusion of thin endometrium can promote the proliferation and repair of damaged endometrium,improve the receptivity of endometrium and reduce the inflammatory reaction of endometrium.

关 键 词：富血小板血浆 薄型子宫内膜 子宫内膜间充质干细胞 子宫内膜容受性 

分 类 号：R711.74[医药卫生—妇产科学]