黄芩对复发性流产孕鼠母胎界面的保护作用

Protective Effect of Scutellaria Baicalensis on Maternal-fetal Interface in RSA Mice and Its Mechanism

目的:探究黄芩是否对复发性流产模型孕鼠母胎界面免疫微环境具有保护作用。方法:正常妊娠模型(正常组)以CBA/J雌鼠×BALB/C雄鼠建立;复发性流产模型以CBA/J雌鼠×DBA/2雄鼠建立,复发性流产模型孕鼠再随机分为模型组和黄芩组,正常组和模型组给予生理盐水,黄芩组给予黄芩水煎剂,连续灌胃14 d,1次/d。14 d后,处死孕鼠,记胚胎数,并计算胚胎丢失率;HE染色后分别观察正常组、模型组及黄芩组胎盘组织的病理改变;透射电镜(TEM)分别观察正常组、模型组及黄芩组的胎盘组织超微结构改变;免疫荧光法测量正常组、模型组及黄芩组白细胞介素6(IL-6)、转化生长因子β1(TGF-β1)表达。结果:与正常组比较,模型组的胚胎丢失率明显上升(P<0.05);与模型组比较,黄芩组胚胎丢失率明显下降(P<0.05);HE染色,正常组胎盘组织结构完整,细胞排列整齐;与正常组比较,模型组出现蜕膜带脱落,巨细胞滋养细胞及海绵滋养层局部坏死,有炎症细胞浸润;与模型组比较,黄芩组未见明显病理改变。TEM观察,正常组胎盘超微结构正常,血管内皮细胞形态结构完整清晰;与正常组比较,模型组胎盘组织超微结构明显异常,合体滋养层细胞、细胞滋养层细胞及血管内皮细胞内线粒体可见不同程度肿胀,并见线粒体自噬小体形成;与模型组比较,黄芩组胎盘组织超微结构较正常,仅见合体滋养层细胞胞浆内出现少量线粒体轻度肿胀。免疫荧光检测提示与正常组比较,模型组孕鼠胎盘组织中IL-6水平上调(P<0.05),TGF-β1水平下调(P<0.05);与模型组比较,黄芩组IL-6水平下调(P<0.05),TGF-β1水平上调(P<0.05)。结论:黄芩可能通过下调IL-6的表达,上调TGF-β1的表达,且调节异常的线粒体自噬过程,介导异常炎性反应,促进免疫耐受,改善胚胎发育进程,使妊娠向良性结局发展,为中医临床应用黄芩安胎提供了理论支持。

Objective:This paper investigates the protective effect of Seutellarla baicalensis on the maternal-fetal interface immune microenvironment in mice with recurrent abortion(RSA)model and then clarifies its mechanism,so as to provide theoretical support for clinical application of Scutellaria baicalensis in fetal safety.Methods:The researcher establishes the normal pregnancy model(normal group)and the recurrent abortion model with CBA/J female rats×BALB/C male rats and CBA/J female rats×DBA/2 male rats respectively.The recurrent abortion model rats are randomly divided into model group and Scutellaria baicalensis group.Normal group and model group are given normal saline,and Scutellaria baicalensis group is given baicalensis decoction once per day for consecutive 14 days.After 14 days,the researcher dissects the mice to observe the number of embryos in the uterus and calculate the rate of embryo loss.Then the researcher observes the pathological changes of placenta were observed through HE staining as well as the ultrastructural changes of placenta by TEM and adopts immunofluorescence assay to dectect the levels of IL-6 and TGF-β1.Results:Compared with normal group,the embryo absorption rate of model group increased(P<0.05),Compared with model group,embryo absorption rate of Scutellaria baicalensis group decreased(P<0.05);HE staining showed that the structure of placental tissue in normal group was intact and cells were orderly arranged.Compared with normal group,the model group showed decidua strip shedding,local necrosis of giant cell trophoblast cells and sponge trophoblast layer,and inflammatory cell infiltration.Compared with model group,no obvious pathological changes were found in Scutellaria baicalensis group.TEM observation shows that the ultrastructure of placenta in normal group was normal,and the morphology and structure of vascular endothelial cells were intact and clear.Compared with the normal group,the ultrastructure of the placental tissues in the model group was obviously abnormal,and the mitochondria in the cytoplasm of syncytiotrophoblast cells,cytotrophoblast cells and vascular endothelial cells were significantly swollen,and the formation of mitochondrial autophagosomes was visible.While in comparison with model group,the ultrastructure of placenta in Scutellaria baicalensis group was not abnormal.Through the Immunofluorescence assay,it shows that IL-6 level in model group is significantly up-regulated(P<0.05)but TGF-β1 level is significantly down-regulated(P<0.05)when compared to normal group.While compared with model group,the level of IL-6 in Seutellarla baicalensis group is significantly decreased(P<0.05)but the level of TGF-β1 is significantly increased(P<0.05).Conclusion:Seutellarla baicalensis has an effective influence on regulating the abnormal inflammatory response,inducing immune tolerance and promoting the development of embryos so as to improve the outcome of pregnancy by the expression of IL-6 down-regulation,the expression of TGF-β1up-regulatio and inhibition of the process of mitochondrial autophagy.It can thus be seen that Seutellarla baicalensis function of miscarriage prevention holds water in the clinical application in traditional Chinese medicine,but its specific mechanism of inhibiting the release of inflammatory factors and mitochondrial autophagy still needs further exploration.