目的 观察清化固肾排毒颗粒对单侧输尿管梗阻大鼠Wnt/β-catenin 信号通路中Wnt1、β 连环蛋白(β-catenin)表达的影响,从而探讨清化固肾排毒颗粒对肾间质纤维化的作用机制。方法 108 只SPF 级SD 大鼠随机分为假手术组和模型组,清化固肾排毒颗粒高、中、低剂量(3.00、1.50、0.75 g/kg)组,洛丁新(1.60 mg/kg)组,每组18 只;除假手术组外,各组大鼠均以结扎左侧输尿管制备单侧输尿管梗阻肾纤维化模型。各组分别于造模第7、14、21 天时随机处死6 只大鼠,采血,检测血清尿素氮(BUN)和肌酐(SCr)水平;取手术侧肾脏组织进行相关病理检测,分别采用HE 染色和Masson 染色评价肾小管间质纤维化损伤程度,免疫组织化学染色检测肾脏组织Wnt1 和β-catenin 的表达情况。结果 与假手术组比较,各造模组大鼠血清SCr、BUN 水平升高(P < 0.05),间质纤维化程度加重(P < 0.05),肾组织β-catenin 表达明显增多(P < 0.05),随着输尿管梗阻时间延长,各指标程度加重;与模型组比较,清化固肾排毒颗粒高、中、低剂量组和洛丁新组大鼠血清SCr、BUN 水平下降(P < 0.05),间质纤维化程度有所缓解(P < 0.05),肾组织β-catenin 表达降低(P < 0.05)。免疫组化结果显示:与假手术组比较,各造模组大鼠肾小管上皮细胞内Wnt1、β-catenin 高表达(P < 0.05),在梗阻后第7 天Wnt1表达量最高(P < 0.05),随后降低;与模型组比较,清化固肾排毒颗粒各剂量组、洛丁新组大鼠Wnt1、β-catenin 表达降低(P < 0.05)。结论 清化固肾排毒颗粒可以改善肾纤维化的发生发展,其作用机制可能是通过抑制Wnt1、β-catenin 的表达,起到保护肾脏的作用。

Objective To investigate the influence and mechaninsm of renal interstitial fibrosis of Qinghua Gushen Paidu Granule (QGPG) on renal tubular epithelial mesenchymal transition in unilateral ureteral obstruction rats. Methods The male Sprague-Dawley (SD) rats (108 cases) were randomly divided into Sham-operated group, model group, high-, mid-, low-dose QGPG groups (HQ, MQ, and LQ groups), and Lotensin treated group, 18 in each group. The rats in HQ, MQ, LQ, and Lotensin treated groups were respectively ig administered with QGPG (3, 1.5, and 0.75 g/kg) and Lotensin (1.6 mg/kg, dissolved in nomal saline). At the same time, nomal saline was ig administered in Sham-opration and model groups, once daily. At days 7, 14, and 21, six rats in each group were randomly sacrificed. Serum blood urea nitrogen (BUN) and serum creatitine (SCr) were tested. Also their left surgery renal tissues were collected for pathological examination. The pathological changes of the obstruction renal tissue were examined by HE and Masson staining. The protein expression of Wnt1 and β-catenin was detected by immunohistochemical staining. Results Compared with the Sham-operation group, serum levels of BUN and SCr of rats in other groups were increased (P < 0.05), protein expression level of β-catenin increased in the renal tissue (P < 0.05). The renal interstitial fibrosis degree was gradually aggratated along with the ureteral obstruction time. Compared with the model group, after the treatment, serum levels of BUN and SCr in rats decreased significantly (P < 0.05), protein expression of β-catenin decreased in the renal tissue (P < 0.05). Compared with the Sham-operation group, the protein expression of Wnt1 in other group increased (P < 0.05). The protein expression of Wnt1 was the highest in day 7 after ureteral obstruction (P < 0.05), then decreased. Compared with the model group, after the treatment by drugs, the protein expression of Wnt1 decreased in the renal tissue (P < 0.05). Conclusion QGPG may ameliorate renal interstitial fibrosis by down-regulating the expression of Wnt1 and β-catenin.

浙江省自然科学基金资助项目(LY14H270001,LY15H280014);浙江省151人才专项;温州市551人才专项;温州市高层次人才课题资助