通脉益肾方拮抗肾间质纤维化的作用机制

Action mechanism of Tongmai Yishen Fang in antagonism of renal interstitial fibrosis

目的观察通脉益肾方对肾间质纤维化模型大鼠肾脏肾上腺髓质素及肾小管上皮细胞表型转化的影响,探讨其拮抗肾间质纤维化的作用机理。方法采用单侧输尿管结扎的方法复制大鼠肾间质纤维化模型,将其随机分为假手术组、模型组、通脉益肾方组和缬沙坦组。通脉益肾方组、缬沙坦组分别给予通脉益肾方1.5 g/(kg.g)和缬沙坦10 mg/(kg.d)灌胃治疗,假手术组及模型组给予等量的生理盐水。观察梗阻侧肾脏肾上腺髓质素(ADM)、肾小管上皮细胞表型转化标志物α-平滑肌肌动蛋白(α-SMA)的蛋白表达,做相关性分析;采用HE及天狼星红染色观察各组大鼠肾脏的病理改变。结果模型组及各治疗组大鼠肾脏中ADM蛋白表达较假手术组明显降低(P<0.05),通脉益肾方组、缬沙坦组的ADM的蛋白含量及表达均高于模型组(P<0.05),但两治疗组之间无显著性差异。模型及各治疗组α-SMA的蛋白表达较假手术组明显升高(P<0.05),通脉益肾方组、缬沙坦组的表达均低于模型组(P<0.05),但两治疗组之间无显著性差异。ADM与α-SMA的表达呈明显负相关。结论通脉益肾方可能通过上调间质纤维化大鼠肾脏ADM的表达、抑制肾小管上皮细胞的表型转化起到拮抗肾间质纤维化的作用。

Objective To observe the influences of Tongmai Yishen Fang on adrenomedullin （ADM） and tubular epithelial phcnotypic transdifferentiation in rats with renal interstitial fibrosis, and to discuss the antagonism mechanism. Methods The rat model of renal interstitial fibrosis was established through unilateral ureteral obstruction, and then all rats were randomly divided into sham-operation group, model group, Tongmai Yishen Fang group and valsartan group. Tongmai Yishen Fang group was given orally Tongmai Yishen Fang [ l. 5 g/（ kg · d） ], valsartan group, valsartan [ 10 nag/（ kg · d） ], and sham- operation group and model group, normal saline water in the same dose. The protein expressions of ADM and α-smooth muscular actin （ α-SMA）, a landmark of tubular epithelial phenotypic transdifferentiation, were observed and analyzed. The pathological changes of kidney were observed after HE staining and Sirius-red staining in rats of all groups. Results The protein expression of ADM decreased significantly in model group, Tongmai Yishen Fang group and valsartan group compared with sham-operation group （P 〈 0.05 ）. The content and protein expression of ADM were higher in Tongmai Yishen Fang group and valsartan group than those in model group （P 〈 0.05 ）, and the difference was not signilicant between two treatment groups. The protein expression of α-SMA increased significantly in model group, Tongmai Yishen Fang group and valsartan group compared with sham-operation group （ P 〈0. 05 ）, and lower significantly in Tongmai Yishen Fang group and valsartan group compared with model group（P 〈0.05 ）, but the difference was not significant between two treatment groups. The expression of ADM and expression of α-SMA showed significant negative correlation. Conclusion Tongmai Yishen Fang can play a role of antagonizing renal interstitial fibrosis through up-reguating the expression of ADM and inhibiting tubular epithelial phenotypic transdifferentiation.