红藤方对子宫内膜异位症大鼠间质-上皮转化的影响
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摘要
目的:探讨中药红藤方对子宫内膜异位症大鼠异位内膜形成包囊过程中间质-上皮转化的影响。方法:采用自体移植法建立大鼠子宫内膜异位症模型,3周后取造模成功的大鼠18只随机分为模型组、红藤方组、孕三烯酮组,分别连续灌胃等容量0.9%Na Cl溶液、红藤方27.72 g·kg~(-1)·d~(-1)、孕三烯酮0.5 mg·kg~(-1)·d~(-1),均持续21 d,结束后取异位内膜组织。采用HE染色方法对异位内膜组织进行病理学观察,免疫组化方法观察上皮标志物上皮细胞钙黏蛋白(E-cadherin)、间质标志物波形蛋白(Vimentin)在异位内膜包囊中的定位和表达情况,Western blot法检测异位内膜包囊形成组织中E-cadherin、Vimentin的蛋白表达。结果:HE染色结果显示,模型组异位内膜组织中腺体减少、间质较多,红藤方组异位内膜组织腔上皮变薄、间质细胞减少,孕三烯酮组异位内膜组织腔上皮完整。免疫组化结果显示,红藤方组和孕三烯酮组异位内膜包囊腔上皮E-cadherin、Vimentin阳性信号较模型组减弱。Western blot结果显示,红藤方组Vimentin和孕三烯酮组E-cadherin蛋白含量较模型组显著降低,差异均有统计学意义(P<0.05)。结论:红藤方可能通过上调间质-上皮转化,导致间质细胞减少,从而抑制子宫内膜异位症大鼠异位内膜包囊的生长。
Objective: To discuss the effects of Hongteng Formula on mesenchymal-epithelial transformation in ectopic endometrium cysts of rats with endometriosis( EMs). Methods: The EMs model was established in rats by autologous transplantation. Three weeks later,18 successfully modeling rats were randomly divided into the model group,Hongteng Formula group and gestrinone group,which treated with 0.9% Na Cl solution at equivalent volume,Hongteng Formula at daily dose of 27. 72 g/kg and gestrinone at daily dose of0.5 mg/kg by intragastric administration for continuous 21 days,respectively. The ectopic endometrium was collected,and the histopathological observation on ectopic endometrium was performed by HE staining. The location and expression of epithelial cadherin( Ecadherin) and Vimentin( as interstitial marker) in the ectopic endometrial cysts were detected by immunohistochemistry. The protein expressions of E-cadherin and Vimentin in the ectopic endometrial cysts were detected by Western blot. Results: The results of HE staining showed that less gland and more mesenchyme were observed in the ectopic endometrium in the model group. The luminal epithelium was thinner and the interstitial cells were decreased in the Hongteng Formula group. The luminal epithelium of ectopic endometrium was complete in the gestrinone group. The results of immunohistochemistry showed that the positive expressions of E-cadherin and Vimentin in the luminal epithelium of ectopic endometrial cysts in the Hongteng Formula group and gestrinone group were lower than those in the model group. The results of Western blot showed that the protein expression of E-cadherin in the gestrinone group and the protein expression of Vimentin in the Hongteng Formula group were significantly lower than those in the model group,with statistically significant differences( P < 0. 05). Conclusion: Hongteng Formula can reduce the mesenchymal cells through mesenchymal-epithelial transformation,and thus inhibit the growth of ectopic endometrial cysts in rats with endometriosis.
Objective: To discuss the effects of Hongteng Formula on mesenchymal-epithelial transformation in ectopic endometrium cysts of rats with endometriosis( EMs). Methods: The EMs model was established in rats by autologous transplantation. Three weeks later,18 successfully modeling rats were randomly divided into the model group,Hongteng Formula group and gestrinone group,which treated with 0.9% Na Cl solution at equivalent volume,Hongteng Formula at daily dose of 27. 72 g/kg and gestrinone at daily dose of0.5 mg/kg by intragastric administration for continuous 21 days,respectively. The ectopic endometrium was collected,and the histopathological observation on ectopic endometrium was performed by HE staining. The location and expression of epithelial cadherin( Ecadherin) and Vimentin( as interstitial marker) in the ectopic endometrial cysts were detected by immunohistochemistry. The protein expressions of E-cadherin and Vimentin in the ectopic endometrial cysts were detected by Western blot. Results: The results of HE staining showed that less gland and more mesenchyme were observed in the ectopic endometrium in the model group. The luminal epithelium was thinner and the interstitial cells were decreased in the Hongteng Formula group. The luminal epithelium of ectopic endometrium was complete in the gestrinone group. The results of immunohistochemistry showed that the positive expressions of E-cadherin and Vimentin in the luminal epithelium of ectopic endometrial cysts in the Hongteng Formula group and gestrinone group were lower than those in the model group. The results of Western blot showed that the protein expression of E-cadherin in the gestrinone group and the protein expression of Vimentin in the Hongteng Formula group were significantly lower than those in the model group,with statistically significant differences( P < 0. 05). Conclusion: Hongteng Formula can reduce the mesenchymal cells through mesenchymal-epithelial transformation,and thus inhibit the growth of ectopic endometrial cysts in rats with endometriosis.
引文
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[7]何钰,付芳华,谷灿灿,等.清热化瘀方治疗血瘀型子宫腺肌病临床观察[J].上海中医药杂志,2015,49(12):52-55.
[8]PROESTLING K,BIRNER P,GAMPERL S,et al.Enhanced epithelial to mesenchymal transition(EMT)and upregulated MYC in ectopic lesions contribute independently to endometriosis[J].Rep Biol Endocrinol,2015,13(1):75-85.
[9]KALLURI R,WEINBERG R A.The basics of epithelial-mesenchymal transition[J].J Clin Invest,2009,119(6):1420-1428.
[10]SCOTTI S,REGIDOR P A,SCHINDLER A E,et al.Reduced proliferation and cell adhesion in endometriosis[J].Mol Hum Reprod,2000,6(7):610-617.
[11]MATSUZAKI S,DARCHA C.Epithelial to mesenchymal transitionlike and mesenchymal to epithelial transition-like processes might be involved in the pathogenesis of pelvic endometriosis[J].Hum Reprod,2012,27(3):712-721.
[12]庄梦斐,张婷婷,孙兆贵,等.清热化瘀法对子宫内膜异位症大鼠血管生成和炎症相关因子表达的影响[J].中华中医药杂志,2016,31(5):1595-1599.
[2]BALDI A,CAMPIONI M,SIGNORILE P G.Endometriosis:pathogenesis,diagnosis,therapy and association with cancer(review)[J].Oncol Rep,2008,19(4):843-846.
[3]竺天虹,张信美.上皮间充质转化介导子宫内膜异位症发生发展的研究进展[J].浙江大学学报(医学报),2016,45(4):439-445.
[4]杨峰,杨凤梅,张婷婷,等.红藤方治疗子宫内膜异位症82例临床观察[J].山东中医药杂志,2006,25(12):824-825.
[5]HAHN D W,CARRAHER R P,FOLDESY R G,et al.Development of an animal model for quantitatively evaluating effects of drugs on endometriosis[J].Fertil Steril,1985,44(3):410-415.
[6]邱琳,邓姗.孕三烯酮治疗子宫内膜异位症效果及不良反应的Meta分析[J].国际妇产科学杂志,2013,40(5):451-463.
[7]何钰,付芳华,谷灿灿,等.清热化瘀方治疗血瘀型子宫腺肌病临床观察[J].上海中医药杂志,2015,49(12):52-55.
[8]PROESTLING K,BIRNER P,GAMPERL S,et al.Enhanced epithelial to mesenchymal transition(EMT)and upregulated MYC in ectopic lesions contribute independently to endometriosis[J].Rep Biol Endocrinol,2015,13(1):75-85.
[9]KALLURI R,WEINBERG R A.The basics of epithelial-mesenchymal transition[J].J Clin Invest,2009,119(6):1420-1428.
[10]SCOTTI S,REGIDOR P A,SCHINDLER A E,et al.Reduced proliferation and cell adhesion in endometriosis[J].Mol Hum Reprod,2000,6(7):610-617.
[11]MATSUZAKI S,DARCHA C.Epithelial to mesenchymal transitionlike and mesenchymal to epithelial transition-like processes might be involved in the pathogenesis of pelvic endometriosis[J].Hum Reprod,2012,27(3):712-721.
[12]庄梦斐,张婷婷,孙兆贵,等.清热化瘀法对子宫内膜异位症大鼠血管生成和炎症相关因子表达的影响[J].中华中医药杂志,2016,31(5):1595-1599.