五苓散干预小鼠肾间质纤维化Wnt/β-catenin信号通路的研究

Wuling Powder Prevents Renal Interstitial Fibrosis in Mice Through Wnt/β-catenin Pathway

目的 探究五苓散(Wuling Powder,WLP)防治小鼠肾间质纤维化(RIF)的机制。方法 将60只小鼠随机分为空白组,模型组,五苓散低、中、高剂量组,采用单侧输尿管梗阻的方法制备小鼠肾间质纤维化模型,空白组及模型组给予生理盐水灌胃,五苓散各组小鼠给予不同剂量五苓散灌胃,连续21 d。检测血清中尿素氮(BUN)和肌酐(Cre)含量;Masson染色观察肾脏组织形态学变化;采用免疫印迹实验(Western Blot)和聚合酶链式反应(PCR)分别检测肾脏组织中Wnt4、β连环蛋白(β-catenin)、结缔组织生长因子(CTGF)mRNA和蛋白的表达水平。结果 与空白组比较,模型组小鼠的血清肌酐、尿素氮含量明显升高(P<0.01);而五苓散低、中、高剂量组的血清尿素氮、肌酐水平下降明显(P<0.05,P<0.01)。模型组小鼠梗阻侧肾脏组织肾间质胶原沉积明显增多(P<0.01);而五苓散干预治疗后,肾间质胶原沉积明显减少(P<0.01)。Western Blot和PCR实验结果显示,与模型组比较,五苓散各剂量组可以不同程度抑制Wnt4、β-catenin和CTGF mRNA和蛋白的表达(P<0.05,P<0.01)。结论 五苓散可以对抗肾间质的纤维化过程,其相关机制可能与抑制肾脏组织中Wnt/β-catenin通路蛋白的表达有关。

Objective To explore the mechanism of Wuling Powder(WLP)in preventing and treating renal interstitial fibrosis(RIF)of mice. Methods Sixty mice were randomly divided into blank group, model group, WLP low-,medium-and high-dose groups. Renal interstitial fibrosis model was prepared by unilateral ureteral obstruction(UUO). The blank group and model group were given normal saline,mice in WLP low-,medium-and high-dose groups were given different doses of WLP. The treatments in all administration groups lasted for 21 days. The contents of blood urea nitrogen(BUN)and serum creatinine(Cre)were determined by kit. Masson staining was used to detect pathological changes of kidney tissue. Western Blot and Real-time PCR were used to detect the protein and mRNA expression levels of Wnt4, β-catenin, and connective tissue growth factor(CTGF) in kidney tissue. Results Compared with the blank group,the levels of Cre and BUN in model group were significantly increased(P<0.01),and the contents of Cre and BUN in WLP low-,medium-and high-dose groups were significantly decreased(P<0.05,P<0.01). In the model group,the collagen deposition of the renal interstitium in the obstructed kidney tissue was significantly increased(P<0.01),and the collagen deposition of renal interstitium was significantly decreased after treatment with WLP(P<0.01). The results of Western Blot and Real-time PCR showed that compared with the model group, different doses of WLP could inhibit the protein and mRNA expressions of Wnt4, β-catenin and CTGF(P<0.05, P<0.01). Conclusion WLP has anti-renal interstitial fibrosis effect, which may be related to inhibiting the expression of Wnt/β-catenin pathway protein in renal tissue.