魏丹丹 张超 朱星昊 梁磊 李闪闪 刘湘花 李真珍 蒋士卿

中圖分类号 R285.5 文献标志码 A 文章编号 1001-0408（2021）08-0915-06

DOI 10.6039/j.issn.1001-0408.2021.08.04

摘 要 目的：研究丹参多酚酸盐对肾间质纤维化（RIF）模型大鼠的改善作用及可能机制。方法：将50只雄性SD大鼠按体质量随机分为正常组、模型组、氯沙坦组（阳性对照组，9 mg/kg）和丹参多酚酸盐低、高剂量组（18、36 mg/kg），每组10只。除正常组外，其余各组大鼠均灌胃腺嘌呤250 mg/kg建立RIF模型。造模结束后，各给药组大鼠灌胃相应药物，正常组和模型组大鼠灌胃等体积生理盐水，灌胃体积均为10 mL/kg，每天1次，连续30天。末次给药后，采用酶联免疫吸附测定法检测大鼠血清中肌酐（Scr）、尿素氮（BUN）和尿液中24 h尿蛋白（24 h UPro）水平;采用苏木精-伊红染色法和Masson染色法分别观察大鼠肾组织病理学特点和纤维化情况，对肾小管损伤、肾小球硬化程度进行评分并计算肾组织阳性染色面积百分比;采用免疫组化法和Western blot法分别测定大鼠肾组织中Wnt蛋白（Wnt5a、Wnt5b）、β-联蛋白（β-catenin）的表达水平。结果：与正常组比较，模型组大鼠Scr、BUN、24 h UPro水平和肾小管损伤评分、肾小球硬化评分、肾组织阳性染色面积百分比以及Wnt5a、β-catenin蛋白的表达水平均显著升高（P<0.05），Wnt5b蛋白的表达水平显著降低（P<0.05）;显微镜下可见大鼠肾组织出现系膜增生、纤维组织增多、炎性细胞浸润等病理学变化。与模型组比较，氯沙坦组和丹参多酚酸盐低、高剂量组大鼠的上述指标均显著改善（P<0.05），且丹参多酚酸盐的作用具有一定的剂量依赖性趋势。结论：丹参多酚酸盐对腺嘌呤致RIF模型大鼠具有改善作用，其机制可能与抑制Wnt/β-catenin信号通路有关。

关键词 丹参多酚酸盐;腺嘌呤;肾间质纤维化;肾功能;Wnt蛋白/β-联蛋白信号通路;大鼠

Study on Improvement Effect of Salvianolate on Renal Interstitial Fibrosis Model Rats and Its Mechanism

WEI Dandan1，ZHANG Chao1，ZHU Xinghao1，LIANG Lei1，LI Shanshan1，LIU Xianghua1，LI Zhenzhen2，JIANG Shiqing1（1. First Clinical College， Henan University of TCM， Zhengzhou 450046， China; 2. Medical Research Center， the First Affiliated Hospital of Zhengzhou University， Zhengzhou 450052， China）

ABSTRACT   OBJECTIVE： To study the improvement effect of salvianolate on renal interstitial fibrosis （RIF） model rats and its possible mechanism. METHODS： Totally 50 male SD rats were randomly divided into normal group， model group， losartan group （positive control group， 9 mg/kg） and salvianolate low-dose and high-dose groups （18， 36 mg/kg） according to body weight， with 10 rats in each group. Except for normal group， other groups were given adenine 250 mg/kg intragastrically to establish RIF model. After modeling， administration groups were given relevant medicine intragastrically， and normal group and model group were given constant volume of normal saline intragastrically， the volume was 10 mL/kg， once a day， for consecutive 30 days. After last medication， the serum levels of creatinine （Scr）， urea nitrogen （BUN） and 24 h proteinuria （24 h UPro） were detected by ELISA. HE staining and Masson staining were used to observe the histopathological characteristics and fibrosis of the kidney. The degree of renal tubular injury and glomerulosclerosis were scored， and the percentage of positive staining area of renal tissue was calculated;immunohistochemistry and Western blot assay were used to determine the protein expression of Wnt5a， Wnt5b， and β-catenin. RESULTS： Compared with normal group， Scr， BUN and 24 h UPro levels， renal tubular injury score，  glomerulosclerosis score， the percentage of positive staining area in renal tissue， the protein expression of Wnt5a and β-catenin were increased significantly in model group （P<0.05）， while the expression of Wnt5b protein was decreased significantly （P<0.05）. Pathological changes such as mesangial hyperplasia， fibrous tissue increase and inflammatory cell infiltration were observed under microscope. Compared with model group， above indexes of rats were improved significantly in losartan group， salvianolate low-dose and high-dose groups （P<0.05）， and the effect of salvianolate had dose-dependent trend. CONCLUSIONS： Salvianolate has the improvement effect on RIF model rats induced by adenine， and its mechanism may be related to inhibition of Wnt/β-catenin signal pathway.

KEYWORDS   Salvianolate; Adenine; Renal interstitial fibrosis; Renal function; Wnt/β-catenin signal pathway; Rat

肾间质纤维化（RIF）是各种病理因素引发的慢性肾病发展至终末期肾衰竭的主要病理特征，主要包括肾血管纤维化、肾小球硬化和肾小管间质纤维化[1-2]。RIF发生的主要环节包括炎性细胞浸润、致纤维化因子激活、肾小管上皮细胞-肾间质肌成纤维细胞分化以及细胞外基质过度沉积等[3]。……