肾安方对慢性肾脏病大鼠Hedgehog信号通路的影响  

Effect of Shen′an Formula(肾安方)on Hedgehog Signal Pathway in Rats with Chronic Kidney Disease

作　　者：宋坚[1,2] 王旭东[1] 郑炜贞[1] 刘宏斌 万洋洋[1] SONG Jian;WANG Xudong;ZHENG Weizhen;LIU Hongbin;WANG Yangyang(Nantong First People′s Hospital,Nantong 226000,Jiangsu,China;Nanjing University of Chinese Medicine,Nanjing 210046,Jiangsu,China)

机构地区：[1]南通市第一人民医院,江苏南通226000 [2]南京中医药大学,江苏南京210046

出　　处：《辽宁中医杂志》2024年第12期192-196,I0021-I0023,共8页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年基金项目(82202436);江苏省中医药科技发展项目(MS2023114);南通市科技局项目(JC2019131)。

摘　　要：目的观察肾安方对慢性肾脏病(chronic kidney disease,CKD)大鼠生化指标和Hedgehog信号通路的影响,探讨其对慢性肾脏病的保护机制。方法60只大鼠随机分为正常组10只和造模组50只,造模组大鼠采用腺嘌呤诱导法复制CKD模型,造模成功的40只大鼠随机分为模型组,肾安方低、高、中剂量组,环巴胺组,各8只。肾安方低、中、高剂量组大鼠分别给予肾安方(12.05、24.1、48.2 g·kg^(-1)·d^(-1))灌胃;环巴胺组大鼠予以环巴胺(6 mg·kg^(-1)·d^(-1))灌胃;正常组和模型组大鼠给予生理盐水等体积灌胃。每日1次,持续给药28 d。第29天,测定各组大鼠血清尿素氮(blood urea nitrogen,BUN)、肌酐(creatinine,Cr)、尿酸(uric acid,UA)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、转化生长因子-β1(transforming growth factor-β1,TGF-β1);HE染色和Masson染色观察肾脏病理变化程度;Western blotting法检测信号通路关键蛋白下游核转录因子GLI家族(GLI1)、Hh分泌型糖蛋白配体(SHH)、成纤维细胞特异蛋白1(fibroblast-specific protein 1,FSP1)、蛋白激酶B(protein kinase B,Akt)、磷酸化蛋白激酶B(p-Akt)蛋白的表达水平;Real-time PCR法检测大鼠肾组织中GLI1mRNA、SHHmRNA、FSP1mRNA、AktmRNA表达水平。结果与模型组比较,肾安方中、高剂量组及环巴胺组BUN、Scr、UA、IL-6、TNF-α、TGF-β1、GLI1mRNA、SHHmRNA、FSP1mRNA、AktmRNA、GLI1、SHH、FSP1、p-Akt及肾脏纤维化程度显著降低(P<0.05);肾安方高剂量组肾功能及炎症指标改善情况与环巴胺组比较差异无统计学意义(P>0.05),但肾安方高剂量组GLI1mRNA、SHHmRNA、FSP1mRNA、AktmRNA、GLI1、SHH、FSP1、p-Akt表达略高于环巴胺组(P<0.05),且肾安方各组间上述指标差异有统计学意义(P<0.05)。结论肾安方可以改善慢性肾脏病大鼠肾功能,可能与抑制Hedgehog信号通路激活有关。Objective To observe the effects of Shen′an Formula(肾安方)on biochemical indicators and Hedgehog signaling pathway in rats with chronic kidney disease(CKD),and explore its protective mechanism against chronic kidney disease.Methods Sixty rats were randomly divided into a normal group(10 rats)and a model group(50 rats).The rats of the model group were induced by adenine to replicate the CKD model.Forty rats that successfully established the model were randomly divided into a model group,low,high and medium dose groups of Shen′an Formula,8 rats in each.The rats in the low,medium and high dose groups of Shen′an Formula were given Shen′an Formula(12.05,24.1,48.2 g·kg^(-1)·d^(-1))by gavage.The rats in the cycloparamide group were given cycloparamide(6 mg·kg^(-1)·d^(-1))by gavage.The rats in the normal group and model group were given an equal volume of normal saline by gavage.The administration was given once a day for 28 days.On day 29,the levels of serum urea nitrogen(BUN),creatinine(Cr),uric acid(UA),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and transforming growth factor-β1(TGF-β1)were measured in each group.HE staining and Masson staining were used to observe the degree of pathological changes in the kidney.Real time PCR was used to detect the expression levels of GLI1mRNA,SHHmRNA,FSP1 mRNA and Akt mRNA in kidney tissue.Western blotting was used to detect the expressions of GLI1,SHH,FSP1,Akt and p-Akt proteins,which were key proteins in the signaling pathway.Results Compared with those of the model group,the middle and high dose groups of Shen′an Formula and the cyclophosphamide group showed the levels of BUN,Scr,UA,IL-6,TNF-α,TGF-β1.GLI1mRNA,SHHmRNA,FSP1mRNA,AktmRNA,GLI1,SHH,FSP1 and p-Akt and renal fibrosis were significantly reduced(P<0.05).There was no statistical significancein the improvement of renal function and inflammatory indicators between the high dose group of Shen′an Formula and the cyclophosphamide group(P>0.05),but the expressions of GLI1mRNA,SHHmRNA,FSP1mRNA,Akt

关 键 词：慢性肾脏病肾 肾脏纤维化 肾安方 HEDGEHOG信号通路 

分 类 号：R285[医药卫生—中药学]