大鼠子宫宫腔粘连模型的建立及评价指标
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摘要
子宫内膜损伤是导致宫腔粘连的最主要原因,建立有效的子宫内膜损伤动物模型是研究此类疾病发生、发展和治疗反应等不可或缺的支撑条件。通过宫腔注射95%的乙醇制造大鼠(Rattus norvegicus)子宫内膜损伤模型,检测胚胎植入数目来分析子宫内膜损伤对大鼠生育情况的影响,观察大鼠子宫内膜厚度、腺体数量和纤维化面积,分析乙醇处理对子宫内膜的影响;检测细胞角蛋白(CK-19)和波性蛋白(Vimentin)的表达水平,分析上皮细胞和间质细胞的增殖分化及子宫内膜损伤程度。结果显示,正常组大鼠子宫比损伤组更为平滑且有韧性,与正常组相比,损伤组大鼠生育率显著降低(P<0.01),子宫内膜厚度变薄(P<0.01)、腺体数量显著减少(P<0.01),纤维化面积显著增大(P<0.01),CK-19和Vimentin表达量显著下调。结果提示已成功建立大鼠子宫内膜损伤动物模型。
Endometrial lesion is the main reason leading to cervical adhesion. Establishing an effective animal model of endometrial injury is an indispensable condition to study the disease occurrence, development and treatment. We established a Rat(Rattus norvegicus) model of endometrial injury by intrauterine injection of 95% alcohol. The numbers of embryo implantation were detected to analyze the effect of endometrial injury on the fertility. The thickness of endometrium, gland numbers and the area of fibrosis were observed to analyze the effects of ethanol treatment on the endometrium. The expressions of cytokeratin 19(CK-19) and Vimentin were detected to analyze the status of epithelium and mesenchymal cells. The results showed that the specimen of Rat uterus in normal group was smooth and showed strong toughness compared with injury group, and the fertility in the injury group was significantly lower than that in control group(P < 0.01)(Fig. 1). Rat endometrium in the injury group was much thinner than that in control group(P < 0.01)(Fig. 2e). Moreover, compared with control group, the gland numbers in the injury group was significantly reduced(P < 0.01)(Fig. 2f), the area of fibrosis was increased(P < 0.01)(Fig. 2g) and the expression of CK-19 and vimentin was remarkably decreased(Fig. 3). The results suggest that the animal model of Rat endometrial injury is successfully established.
Endometrial lesion is the main reason leading to cervical adhesion. Establishing an effective animal model of endometrial injury is an indispensable condition to study the disease occurrence, development and treatment. We established a Rat(Rattus norvegicus) model of endometrial injury by intrauterine injection of 95% alcohol. The numbers of embryo implantation were detected to analyze the effect of endometrial injury on the fertility. The thickness of endometrium, gland numbers and the area of fibrosis were observed to analyze the effects of ethanol treatment on the endometrium. The expressions of cytokeratin 19(CK-19) and Vimentin were detected to analyze the status of epithelium and mesenchymal cells. The results showed that the specimen of Rat uterus in normal group was smooth and showed strong toughness compared with injury group, and the fertility in the injury group was significantly lower than that in control group(P < 0.01)(Fig. 1). Rat endometrium in the injury group was much thinner than that in control group(P < 0.01)(Fig. 2e). Moreover, compared with control group, the gland numbers in the injury group was significantly reduced(P < 0.01)(Fig. 2f), the area of fibrosis was increased(P < 0.01)(Fig. 2g) and the expression of CK-19 and vimentin was remarkably decreased(Fig. 3). The results suggest that the animal model of Rat endometrial injury is successfully established.
引文
Bargagna-Mohan P,Lei L,Thompson A,et al.2015.Vimentin phosphorylation underlies myofibroblast sensitivity to withaferin a in vitro and during corneal fibrosis.PLo S One,10(7):e0133399.
Berman J M.2008.Intrauterine adhesions.Seminars in Reproductive Medicine,26(4):349-355.
Conforti A,Alviggi C,Mollo A,et al.2013.The management of Asherman syndrome:a review of literature.Reproductive Biology and Endocrinology,11(1):118.
Polishuk W Z.1975.Endometrial regeneration and adhesion formation.Samj South African Medical Journal,49(12):440-442.
Yang X,Zhang M,Zhang Y,et al.2011.Mesenchymal stem cells derived from Wharton jelly of the human umbilical cord ameliorate damage to human endometrial stromal cells.Fertility and Sterility,96(4):1029-1036.e4.
Yu D,Wong Y M,Cheong Y,et al.2008.Asherman syndrome-one century later.Fertility and Sterility,89(4):759-779.
陈学忠,舒沪英,钱振坤,等.1956.活血化瘀方防治子宫内膜粘连的实验研究.中西医结合杂书,6(12):740-742.
高红,赵静,李艳萍.2011.大鼠薄型子宫内膜模型的建立和鉴定生命科学研究,15(5):426-431.
曲军英,吕一帆,林茵,等.2011.建立小鼠子宫内膜损伤模型的研究.福建医科大学学报,45(1):34-36,44.
项华,章晓玲,查捷,等.2008.SD大鼠生长发育指标和主要脏器正常参考值的探讨.中国卫生检验杂志,18(2):299-301.
徐慧颖,李娜,张云山.2014.胚胎植入--子宫内膜容受性是关键.生殖医学杂志,23(3):198-202.
俞凌.2012.人脐带间充质干细胞治疗卵巢早衰的初步实验研究北京:解放军总医院军事进修学院博士学位论文.
Berman J M.2008.Intrauterine adhesions.Seminars in Reproductive Medicine,26(4):349-355.
Conforti A,Alviggi C,Mollo A,et al.2013.The management of Asherman syndrome:a review of literature.Reproductive Biology and Endocrinology,11(1):118.
Polishuk W Z.1975.Endometrial regeneration and adhesion formation.Samj South African Medical Journal,49(12):440-442.
Yang X,Zhang M,Zhang Y,et al.2011.Mesenchymal stem cells derived from Wharton jelly of the human umbilical cord ameliorate damage to human endometrial stromal cells.Fertility and Sterility,96(4):1029-1036.e4.
Yu D,Wong Y M,Cheong Y,et al.2008.Asherman syndrome-one century later.Fertility and Sterility,89(4):759-779.
陈学忠,舒沪英,钱振坤,等.1956.活血化瘀方防治子宫内膜粘连的实验研究.中西医结合杂书,6(12):740-742.
高红,赵静,李艳萍.2011.大鼠薄型子宫内膜模型的建立和鉴定生命科学研究,15(5):426-431.
曲军英,吕一帆,林茵,等.2011.建立小鼠子宫内膜损伤模型的研究.福建医科大学学报,45(1):34-36,44.
项华,章晓玲,查捷,等.2008.SD大鼠生长发育指标和主要脏器正常参考值的探讨.中国卫生检验杂志,18(2):299-301.
徐慧颖,李娜,张云山.2014.胚胎植入--子宫内膜容受性是关键.生殖医学杂志,23(3):198-202.
俞凌.2012.人脐带间充质干细胞治疗卵巢早衰的初步实验研究北京:解放军总医院军事进修学院博士学位论文.