基于Rho/Rock通路的三七茜草有效组分宫腔给药对IUD模型大鼠子宫血管平滑肌细胞表型转化机制研究

Effects of Intrauterine Administration of Effective Components of Panax Notoginseng and Radix Rubiae on Phenotype and Rho/Rock Pathway of Uterine Vascular Smooth Muscle Cells in IUD Rats

目的通过三七茜草有效组分宫腔给药对IUD(Intrauterine device)模型大鼠进行干预,观察相关指标以探讨其对子宫血管平滑肌细胞(Vascular smooth muscle cell,VSMC)的表型转化影响的机制。方法选用40只SPF级SD雌性已生育大鼠,随机分为空白对照组(空白组)、宫内节育器组(IUD组)、三七茜草有效组分宫腔给药组(SQ-IUD组)、宫宁颗粒灌胃组(宫宁颗粒组)各10只,观察大鼠精神活动状态、子宫炎症水肿情况,电镜下观察子宫内膜形态学改变,ELISA检测宫腔冲洗液血小板源性生长因子(Plateletderived growth factor,PDGF)含量,免疫组化法检测RhoA蛋白表达水平,PCR法检测RhoA,Rho激酶Ⅱ(ROCKⅡ)、平滑肌22 alpha(SM22α)的mRNA含量。结果在大鼠造模后状态方面,宫宁颗粒组与SQ-IUD组一般状态与空白组无差别,子宫炎症水肿情况较IUD组轻;在PDGF方面,SQ-IUD组含量显著低于IUD组(P<0.05);在免疫组化方面,SQ-IUD组RhoA蛋白表达显著低于IUD组、宫宁颗粒组(P<0.01);在RhoA mRNA方面,SQ-IUD组含量显著低于IUD组、宫宁颗粒组(P<0.01);在RockⅡmRNA方面,SQ-IUD组含量显著低于IUD组(P<0.05),但与宫宁颗粒组差异无统计学意义(P>0.05);在SM22αmRNA方面,SQ-IUD组含量显著高于IUD组、宫宁颗粒组(P<0.05)。结论三七茜草有效组分宫腔给药可明显减轻IUD放置后引起的炎症反应,可能通过抑制Rho/Rock信号通路激活减少子宫VSMC的表型转化,从而起到防治IUD出血不良反应。

Objective To investigate the mechanism of the effect of Panax notoginseng and Radix Rubiae on the phenotypic transformation in uterine vascular smooth muscle cells(VSMCs)by intrauterine administration of Panax notoginseng effective components in intrauterine device(IUD)model rats.Methods A total of 40 SPF SD female fertile rats were randomly divided into blank control group(n=10),IUD group(n=10),Panax notoginseng effective component intrauterine administration group(SQ-IUD group),and Gongning granule intragastric administration group(n=10).The mental activity,uterine inflammation,and edema were observed,and the morphological changes of the endometrium were observed under the electron microscope.The content of platelet-derived growth factor(PDGF)in uterine lavage fluid was detected by ELISA.The expression of RhoA protein was detected by immunohistochemical method,and the contents of RhoA mRNA,RockII and SM22αmRNAs were detected by PCR method.Results There was no difference in the general state between Gongning granule group and SQ-IUD group and blank group,uterine inflammation,and edema in Gongning granule group and SQ-IUD group was slighter than that in IUD group,the content of PDGF in SQIUD group was significantly lower than that in IUD group,and the expression of RhoA protein in SQ-IUD group was significantly lower than that in IUD group and Gongning granule group(P<0.05).The content of RhoAmRNA in SQIUD group was significantly lower than that in IUD group and Gongning granule group,and the content of RockⅡmRNA in SQ-IUD group was significantly lower than that in IUD group,but there was no significant difference between SQ-IUD group and Gongning granule group(P>0.05).The content of SM22αmRNA in SQ-IUD group was significantly higher than that in IUD group and Gongning granule group.Conclusion Intrauterine administration of Panax notoginseng and Radix Rubiae can significantly reduce the inflammatory reaction caused by the placement of IUD and may reduce the phenotypic transformation of uterine VSMC by inhibiting the activation of Rho/Rock signal pathway,thus preventing and treating the adverse reactions of IUD bleeding.