益肾软坚散对慢性马兜铃酸肾病大鼠模型肾间质纤维化的保护作用

Protective Effects of Yishen Ruanjian Power on Renal Interstitial Fibrosis in Chronic Aristolochic Acid Induced Nephropathy Rat Model

目的研究中药方剂益肾软坚散能否对慢性马兜铃酸肾病(CAAN)大鼠的肾损害起到保护作用。方法雄性SD大鼠18只分为3组,每组6只。模型组予关木通浸膏水溶液间断灌胃制成CAAN动物模型,中药组在造模基础上同步予益肾软坚散水煎剂灌胃,对照组予自来水灌胃。于1、4、8、12、16周分别检测大鼠体重、24h尿蛋白定量、尿β2微球蛋白、血清肌酐(SCr);并于第16周末处死大鼠,取肾脏组织,切片做Masson染色观察肾脏病理变化;用RT-PCR及免疫组化方法检测肾组织中转化生长因子-β1(TGF-β1)、结缔组织生长因子(CTGF)、纤溶酶原激活物抑制物-1(PAI-1)、金属蛋白酶组织抑制物-1(TIMP-1)和Ⅰ型胶原(ColⅠ)mRNA及蛋白质表达。结果与对照组比较,模型组大鼠24h尿蛋白定量、尿β2微球蛋白、SCr自造模后1周出现不同程度的上升(P<0.01,P<0.05);16周时肾间质纤维化面积显著扩大(P<0.01);肾组织内TGF-β1、CTGF、PAI-1、TIMP-1及ColⅠ各指标的mRNA及蛋白质表达均显著上调(P<0.01)。中药干预后,上述上调指标除24h尿蛋白定量外均被显著抑制(P<0.01,P<0.05)。结论益肾软坚散可抑制肾间质内细胞外基质蓄积,从而改善CAAN的肾间质纤维化及肾功能。

Objective To study the protective effects of Yishen Ruanjian Power （YRP） on renal interstitial fibrosis in rats with chronic aristolochic acid induced nephropathy （CAAN）. Methods Eighteen male SD rats were divided into 3 groups, 6 in each group. Water solution of Caulis Aristolochia Manshuriensis （CAM） Liquid Extract were given to the mice in the model group by gastrogavage to make CAAN animal model. For those in the TCM group, decocted water solution of YRP was given by gastrogavage after the mice being modeled with the above-mentioned method. Tap water was given by gastrogavage to the mice in the control group. Body weight, 24-hr urinary protein excretion and/32 microglobulin （/32-MG）, and serum creatinine （SCr） were determined at the end of the 1st, 4th, 8th, 12th and 16th week. At the end of the 16th week, the rats were sacrificed and the pathological figure of their kidneys were observed by Masson staining. Transforming growth factor-β1 （TGF-β1 ）, connective tissue growth factor （CTGF）, plasminogen activator inhibitor-1 （PAI-1）, tissue inhibitor of metalloproteinase-1 （TIMP-1） and type Ⅰ collagen （Col Ⅰ ） in kidney tissue were determined by RT-PCR and immunohistochemical method, respectively. Results At end of the 1st week, urinary protein excretion, urinary β2-MG and SCr in the model group were significantly increased to the levels higher than those in the control group （P 〈 0.01 or 0.05）. Relative area of interstitial fibrosis was significantly enlarged in the model group at the end of the 16th week （P〈0.01）, and at the same time, the mRNA and protein expression of TGF-/31 , CTGF, PAI-1, TIMP-1 and Col Ⅰ in kidney tissue were significant up-regulated （ P 〈 0.01 ）. After intervention with YRP, the above-mentioned up-regulated parameters, except 24-hr urinary protein excretion, were all significantly inhibited （ P 〈 0.01 or 0.05）. Conclusion YRP could inhibit the accumulation of extracellular matrix in renal interstitial tissue, so as to alleviate the renal interstitial fibrosis and improve the renal function.