基于内皮/上皮-间质转化探讨瓜蒌薤白汤治疗Ⅱ型心肾综合征的作用机制

Effect of Gualou Xiebai Decoction on typeⅡcardiorenal syndrome based on endothelial/epithelial-to-mesenchymal transition and its mechanism

目的观察不同剂量瓜蒌薤白汤(Gualou Xiebai Decoction,GXD)对Ⅱ型心肾综合征(typeⅡcardiorenal syndrome,typeⅡCRS)的保护作用,并探讨其初步作用机制。方法通过结扎左冠状动脉前降支复制Ⅱ型心肾综合征大鼠模型,灌胃给药10周后,超声心动图评估各组大鼠的心脏功能;检测血清中生化指标;HE、Masson染色分别观察心肾组织病理改变;碱水解法测定心、肾组织中的羟脯氨酸含量;免疫荧光法、Western blot法检测心脏、肾脏组织中内皮/上皮-间质转化相关蛋白的表达情况。结果GXD低、高剂量组动物心、肾组织脏器指数明显降低(P<0.05,P<0.01),心脏舒张收缩功能指标均有明显提升(P<0.05,P<0.01);血清生化指标AST、LDH、CK-MB、UA、Cr和BUN均有明显改善(P<0.01);心、肾组织的病变程度明显改善,羟脯氨酸含量明显减少(P<0.01);心、肾组织CD31、E-cadherin蛋白的表达水平明显上调(P<0.05,P<0.01),Vimentin、α-SMA和TGF-β1蛋白的表达水平明显下调(P<0.05,P<0.01)。结论GXD对结扎左冠状动脉前降支所致Ⅱ型心肾综合征大鼠的心肾功能有明显的改善作用,延缓心、肾纤维化的进程,其机制可能与抑制内皮/上皮-间质转化有关。

Aim To investigate the protective effects of different doses Gualou Xiebai Decoction(GXD)on typeⅡcardiorenal syndrome(typeⅡCRS)and explore its preliminary mechanisms.Methods The typeⅡcardiorenal syndrome rat model was replicated by ligating the left anterior descending coronary artery.After 10 weeks of intragastric administration,the cardiac function of the rats in each group was evaluated by echocardiography;serum were collected for biochemical testing;heart and kidney tissue samples were stained with HE and Masson to observe pathological changes.The hydroxyproline content in the heart and kidney was detected.The expression levels of endothelial/epithelial-to-mesenchymal transition(EndMT/EMT)related proteins in heart and kidney tissues were detecterd by immunofluorescence double staining、Western blot.Results The index of heart and kidney organs of the low and high doses of GXD group significantly decreased(P<0.05,P<0.01),the cardiac function indexes were significantly improved(P<0.05,P<0.01).Serum characteristic indexes showed that heart and kidney functions were significantly improved(P<0.01).In addition,the pathological changes of the heart and kidney were significantly ameliorated.The hydroxyproline content in heart and kidney correspondingly declined(P<0.01).The expression levels of CD31 and E-cadherin were significantly up-regulated(P<0.05,P<0.01),and the expression levels of Vimentin,α-SMA and TGF-β1 were down-regulated in heart and kidney tissues(P<0.05,P<0.01).Conclusions Therefore,GXD can significantly improve the heart and kidney function of typeⅡCRS rats caused by ligation of the left anterior descending coronary artery,and delay the process of heart and kidney fibrosis.The mechanism may be related to the inhibition of EndMT/EMT.