慢肾康宁对腺嘌呤致肾间质纤维化大鼠肾组织TGF-β1/Smad3/Snail1通路的影响  

Manshenkangning Has Effects on TGF-β1/Smad3/Snail1 Signaling Pathway in Kidney Tissue of Rats with Renal Interstitial Fibrosis Due to Adenine

作　　者：葛茂功 孙升云[2] 张静 朱诗平[2] 冯伟峰[2] GE Maogong;SUN Shengyun;ZHANG Jing;ZHU Shiping;FENG Weifeng

机构地区：[1]河南医学高等专科学校附属医院,河南郑州451191 [2]暨南大学附属第一医院,广东广州510632 [3]广州市天河区棠下街社区卫生服务中心,广东广州510632

出　　处：《新中医》2022年第7期1-7,共7页New Chinese Medicine

基　　金：广东省科技计划项目(2014A020221015)。

摘　　要：目的:观察慢肾康宁对腺嘌呤致肾间质纤维化(RIF)大鼠肾组织中转化生长因子-β1 (TGF-β1)、Smad3、Snail1、E-钙黏着蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)表达水平的影响。方法:将60只Wistar大鼠随机分为对照组、模型组、氯沙坦组及慢肾康宁高、中、低剂量组,每组10只。除对照组外,其余各组制备腺嘌呤致RIF大鼠模型。模型制备成功后,对照组及模型组灌胃蒸馏水,氯沙坦组按10 mg/(kg·d)的剂量灌胃氯沙坦混悬液,高、中、低剂量组分别按30、15、7.5 mg/(kg·d)的剂量灌胃慢肾康宁混悬液,均灌胃30 d。采用HE染色和Massan染色观察肾组织病理变化;免疫组织化学法测定E-cadherin、α-SMA、TGF-β1蛋白表达水平;采用蛋白质免疫印迹法检测肾组织中Smad3、Snail1蛋白表达水平;采用全自动生化分析仪检测血肌酐(SCr)、尿素氮(BUN)、24小时尿蛋白定量(24 h MTP)、肾小球滤过率(eGFR)水平。结果:与模型组比较,氯沙坦及慢肾康宁组SCr、BUN、24 h MTP水平均显著下降,e GFR上升;病理观察,模型组可见肾小管萎缩或扩张,炎症细胞浸润明显,肾小球及肾间质内可见腺嘌呤代谢产物聚集及蓝染胶原纤维聚集,氯沙坦及慢肾康宁各组中病理表现相对较轻;与模型组比较,各给药组α-SMA、TGF-β1、Snail1蛋白表达水平下降、E-cadherin蛋白表达水平升高,氯沙坦及慢肾康宁中、高剂量组中Smad3蛋白表达水平明显下降。结论:慢肾康宁能够抑制RIF大鼠TGF-β1/Smad3/Snail1信号传导,增加E-cadherin、拮抗α-SMA蛋白的表达,具有肾脏保护作用,这可能是其抗RIF的作用机制之一。Objective:To observe the effect of Manshenkangning on expression levels of transforming growth factor-β1(TGF-β1),Smad3,Snail1,E-cadherin,α-smooth muscle actin(α-SMA) in the kidney tissue of rats with renal interstitial fibrosis(RIF) due to adenine. Methods:A total of 60 Wistar rats were randomly divided into the control group,the model group,the losartan group,and the Manshenkangning groups of high-dose,medium-dose and low-dose,with 10 rats in each group. Except for the control group, the rat model of RIF induced by adenine was prepared in other groups. After successful modeling,the control group and the model group were given gastric perfusion with distilled water;the losartan group was given gastric perfusion with losartan suspension at the dose of 10 mg/(kg·d);the groups of high-dose,mediumdose and low-dose were given gastric perfusion with Manshenkangning suspension at the doses of 30,15 and 7.5 mg/(kg·d)respectively;all the groups were perfused for 30 days. The pathological changes of kidney tissue were observed by HE staining and Massan staining;expression levels of E-cadherin, α-SMA, and TGF-β1 protein were determined by immunohistochemistry;the expression levels of Smad3 and Snail1 protein in kidney tissue were detected by Western blot method;the levels of serum creatinine(SCr), blood urea nitrogen(BUN), 24-hour urine protein quantity(24 h MTP) and estimated glomerular filtration rate(eGFR) were measured by automatic biochemical analyzer. Results:When compared with those in the model group,levels of SCr,BUN and 24 h MTP in the losartan group and the Manshenkangning groups were significantly decreased, and levels of eGFR were increased. According to pathological observation, there were tubular atrophy or expansion, obvious infiltration of inflammatory cells, accumulation of adenine metabolite products and bluestained collagen fibers in the glomerulus and renal interstitium in the model group,and the pathological manifestations in the losartan group and the Manshenkangning groups were relatively mil

关 键 词：肾间质纤维化 慢肾康宁 腺嘌呤 TGF-β1/Smad3/Snail1信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]