【摘要】 目的 探讨芪黄补肾泄浊方对UUO大鼠模型Notch信号通路表达的影响，探讨其延缓慢性肾衰竭(CRF)肾间质纤维化(RIF)进展的机制。方法 将80只SPF级雄性SD大鼠随机分为假手术组、模型组、芪黄补肾泄浊方组(芪黄补肾组)与厄贝沙坦组，每组20只。采用左侧输尿管完全梗阻法(UUO)建立慢性肾衰竭肾间质纤维化模型，大鼠灌胃，芪黄补肾泄浊方组及厄贝沙坦组给予相应药物水溶液灌胃，假手术组与模型组给予等体积蒸馏水灌胃，每日1次。观察大鼠一般状态；收集大鼠尿液、血清进行尿量、24hUPQ、血清肌酐、血清尿素氮检测；收集各组大鼠梗阻侧肾组织，HE、Masson染色观察肾脏病理改变。免疫组化法观察Notch1、Jagged1和Hes1蛋白表达；Real-Time PCR法检测Notch1 mRNA、Jagged1 mRNA、Hes1 mRNA基因转录；Western blot法检测Notch1、Jagged1、Hes1蛋白表达。结果 与假手术组相比，模型组大鼠状态差，皮毛暗淡，反应迟钝，活动减少，喜蜷卧，饮食量明显减少，芪黄补肾泄浊方可以改善UUO大鼠一般状态，且优于厄贝沙坦组。模型组大鼠24hUPQ、血清肌酐及血清尿素氮与假手术组相比明显升高(P<0.05);芪黄补肾泄浊方及厄贝沙坦可降低UUO大鼠24hUPQ、血清肌酐及血清尿素氮水平(P<0.05)。HE及Masson染色结果显示：与模型组比，芪黄补肾泄浊方可改善UUO大鼠肾脏的病理变化程度，可减少UUO大鼠肾组织纤维化面积百分比(P<0.05)。免疫组化、Real-Time PCR、Western blot结果显示：在大鼠肾组织中，与假手术组相比，Notch1、Jagged1、Hes1蛋白及mRNA表达在模型组明显增加(P<0.05),在芪黄补肾组、厄贝沙坦组中的表达均少于模型组(P<0.05),且芪黄补肾组少于厄贝沙坦组(P<0.05)。结论 芪黄补肾泄浊方可改善肾功能，延缓慢性肾衰竭肾间质纤维化，其作用可能与降低Notch1mRNA、Jagged1mRNA、Hes1mRNA基因转录，下调Notch1、Jagged1、Hes1蛋白表达，从而抑制Notch信号通路有关。更多还原

【Abstract】 Objective The purpose of this study was to investigate the effect of Qihuang Bushen Xiezhuo Formula on the expression of Notch signaling pathway in UUO rat model and its mechanism of delaying the progression of renal interstitial fibrosis(RIF) in chronic renal failure(CRF). Methods A total of 80 SPF-grade male SD rats were randomly divided into a sham operation group, a model group, a Qihuang Bushen Xiezhuo Formula group(Qihuang Bushen group) and an irbesartan group, 20 rats in each group.After the left complete ureteral obstruction(UUO) was used to establish the renal interstitial fibrosis model of chronic renal failure, the rats were intragastrically administered with the corresponding aqueous solutions of drugs in the Qihuang Bushen Xiezhuo Formula group and irbesartan group. The rats in the sham operation group and the model group were intragastrically administered with the same volume of distilled water once a day. Then we observe that general state of rat; Urine and serum were collected for determination of urine volume, 24hUPQ, serum creatinine and serum urea nitrogen. The tissues of the obstructed kidney were collected from each group, and the pathological changes of the kidney were observed by HE and Masson staining. The expressions of Notch1, Jagged1 and Hes1 proteins were observed by immunohistochemistry. The transcription of Notch1 mRNA, Jagged1 mRNA and Hes1 mRNA was detected by Real-Time PCR. The expressions of Notch1, Jagged1 and Hes1 proteins were detected by Western blot.Results Compared with the sham operation group, the rats in the model group had poor condition, dark fur, slow reaction, reduced activity, like curled up and lying down, and significantly reduced diet. Qihuang Bushen Xiezhuo Formula could improve the general state of UUO rats, and the effect was better than that of irbesartan group.The 24hUPQ, serum creatinine and serum urea nitrogen levels in the model group were significantly higher than those in the sham operation group(P < 0.05). Qihuang Bushen Xiezhuo Formula and irbesartan could reduce the level of 24hUPQ, serum creatinine and serum urea nitrogen in UUO rats(P < 0.05). HE and Masson staining results showed that compared with the model group, Qihuang Bushen Xiezhuo Formula could improve the degree of pathological changes in the kidney of UUO rats and reduce the percentage of renal fibrosis area in UUO rats(P < 0.05). Immunohistochemical, Real-Time PCR and Western blot results showed that compared with the sham operation group, the protein and mRNA expression levels of Notch1, Jagged1, Hes1 in the model group were significantly increased(P < 0.05). The expression levels in the Qihuang Bushen group and irbesartan group were less than that in the model group(P < 0.05). The expression levels of Notch 1, Jagged 1, Hes1 in the Qihuang Bushen group were less than that in the irbesartan group(P < 0.05). Conclusion Qihuang Bushen Xiezhuo Formula can improve renal function and delay renal interstitial fibrosis in CRF patients. Its effect may be related to reducing the transcription of Notch1mRNA, Jagged1mRNA and Hes1mRNA genes, down-regulating the expression of Notch1, Jagged1 and Hes1 proteins, and thus inhibiting Notch signaling pathway.更多还原