肾衰灵方作用于SHH信号通路对UUO大鼠肾间质纤维化的影响  

Effect of Shen Shuai Ling Decoction on SHH Signal Pathway in Renal Interstitial Fibrosis in UUO Rats

作　　者：骆言 刘洪[1] 黎颖[1] 宋娜[2] 熊维建[1] 张玲[1] 丁伟森 Luo Yan;Liu Hong;Li Ying;Song Na;Xiong Weijian;Zhang Ling;Ding Weisen(Department of Nephrology Chongqing Trandition Chinese Medicine Hospital,Chongqing 400021,China;Department of Oncology Chongqing Traditional Chinese Medicine Hospital,Chongqing 400021,China)

机构地区：[1]重庆市中医院肾病科,重庆400021 [2]重庆市中医院肿瘤科,重庆400021

出　　处：《世界科学技术-中医药现代化》2020年第8期3016-3022,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会青年科学基金项目(81904012):基于SHH-Gli-FoxM1途径研究肾衰灵方减轻肾间质纤维化的作用机制,负责人:黎颖;重庆市科学技术局自然科学基金项目(cstc2018jcyjAX0783):肾衰灵方上调PLZF表达减轻肾间质纤维化的体外机制研究,负责人:黎颖;重庆市科学技术局自然科学基金项目(cstc2017jcyjAX0465):肾衰灵方抑制SHH信号通路减轻大鼠肾间质纤维化的机制研究,负责人:黎颖

摘　　要：目的探讨当代名医国医大师郑新经验方—肾衰灵方对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的抑制作用。方法1.50只雄性SD大鼠随机分为五组:假手术组、模型组、肾衰灵方低剂量组、肾衰灵方中剂量组、肾衰灵方高剂量组。除假手术组,其余各组结扎单侧输尿管建立肾间质纤维化动物模型,肾衰灵方低中高剂量组予肾衰灵方(5、15、25 mg·kg-1)连续灌胃14天。检测各组大鼠血清肌酐(SCr)、尿素氮(BUN)、谷丙转氨酶(ALT),谷草转氨酶(AST),碱性磷酸酶(ALP),乳酸脱氢酶(LDH)。采用real-time PCR检测结缔生长因子(connective tissue growth factor,CTGF)核酸水平,采用Masson染色方法检测肾组织观察肾小管和间质病理形态变化。2.根据结果选择肾衰灵中剂量组为治疗组,再将30只雄性SD大鼠分为假手术组、模型组、肾衰灵方治疗组共3组。用Western blot方法检测音猬因子(Sonic hedgehog,SHH)、神经胶质瘤相关癌基因同源蛋白-1(Glioma related cancer gene homologous proteins 1,Gli1)、锌指转录因子(Snail Family Zinc Finger 1,Snail)蛋白的表达。结果与模型组相比,模型组和肾衰灵各剂量组大鼠血清SCr和BUN的表达水平均明显下调,其中肾衰灵中剂量组疗效最佳(P<0.05);Masson染色发现相较模型组,肾衰灵方各剂量组肾间质纤维化明显减轻,其中肾衰灵中剂量组疗效最佳(P<0.05);real-time PCR结果发现肾衰灵各剂量组CTGF核酸水平均较模型组下调,其中肾衰灵中剂量组核酸水平最低(P<0.05)。相较假手术组,模型组与肾衰灵方组SHH、Gli1、Snai1蛋白的表达均升高(P<0.05);与模型组相比,肾衰灵方组SHH、Gli1、Snai1蛋白水平明显降低(P<0.05)。结论单侧输尿管梗阻(UUO)大鼠模型中,肾衰灵方可以通过抑制肾间质纤维化改善大鼠肾功能,其作用可能与其阻断SHH信号通路相关。Objective To explore the inhibitory effect of ShenShuaiLing Decoction(SSLF),the experienced physician of contemporary famous doctor ZhengXin’s empirical prescription,on renal interstitial fibrosis.Methods 1.50 male SD rats were randomly divided into 5 groups:sham,model,SSLF-LOW,SSLF-Mid,SSLF-High,10 rats in each group.Unilateral ureteml obstmction(UUO)model rats,except the sham group,were established by ligating left ureter.The sham and model group were treated with equal volume of normal saline.The SSLF groups were lavaged with ShenShuaiLing Decoction(5,15,25 mg·kg-1)once daily.Serum creatinine SCr and BUN,ALT,AST,ALP,LDH of rats in each group were detected.The connective growth factor(connective tissue growth factor,CTGF)was detected by realtime PCR.The morphological change and the collagen fiber was observed by Masson staining.2.According to the results,30 male SD rats were divided into sham,model,SSLF groups.The expression of sonic hedgehog(SHH),glioma related cancer gene homologous proteins 1(Gli1)and Transcriptional factor Snail was detected by Western blot method.Result 1.Compared with model group,the SCr and BUN expressions of each dose SSLF group were significantly decreased remarkably(P<0.05),and the renal interstitium showed the fibrotic lesions in model group more remarkably than SSLF groups(P<0.05).The CTGF mRNA level of each dose group of SSLF was down-regulated compared with that of the model group(P<0.05).The effect of SSLF medium dose group was the best.Compared with Model group,the SCr,BUN,and the expression of SHH,Glil and Snail in SSLF groups were significantly decreased remarkably(P<0.05).Conclusion SSLF could remarkably improve renal function and alleviates renal interstitial fibrosis of UUO rats.And its possible mechanism may have a relationship with inhibiting activating SHH signal pathway.

关 键 词：肾衰灵方 UUO模型大鼠 肾间质纤维化 SHH信号通路 

分 类 号：R285.5[医药卫生—中药学]