温肾健脾活血泄浊方对慢性肾脏病大鼠肾脏转化生长因子-β1-Smad2/3信号通路的作用  

Effect of warming kidney and strengthening spleen and promoting blood circulation and eliminating turbidity decoction on TGF-β1-Smad2/3 signaling pathway in kidney tissue of rats with chronic kidney disease

作　　者：严胜利 谢娜 舒占钧[2] YAN Shengli;XIE Na;SHU Zhanjun(Department of Traditional Chinese Medicine Surgery,the First Affiliated Hospital of Shihezi University,Shihezi 832000,China)

机构地区：[1]石河子大学第一附属医院中医外科,832000 [2]新疆医科大学附属中医医院中医药临床研究基地

出　　处：《环球中医药》2024年第11期2212-2218,共7页Global Traditional Chinese Medicine

基　　金：石河子大学科研项目(ZZZC2022098)。

摘　　要：目的探讨金洪元教授温肾健脾活血泄浊方对慢性肾脏病(chronic kidney disease,CKD)实验大鼠的肾脏组织内转化生长因子(transforming growth factor,TGF)-β1-Smad2/3信号通路的作用。方法将56只大鼠随机分为空白组13只,其余43只大鼠灌服腺嘌呤溶液制备CKD大鼠模型,经造模鉴定后,取10只空白组大鼠作为空白对照组,40只造模成功大鼠随机分为模型组、中药低、中、高剂量组,每组10只。中药低、中、高剂量组分别给予的灌服浓度为9.18 g/(kg·d)、18.36 g/(kg·d)、36.72 g/(kg·d),空白对照组及模型组大鼠均灌服等体积生理盐水,连续灌服4周。经干预后,观察大鼠一般情况;运用酶联免疫吸附法检测大鼠尿液24小时尿蛋白定量、肌酐、TGF-β1及血清TGF-β1、胱抑素-C及肌酐定量水平;苏木精—伊红染色、马松(Masson)染色观察大鼠肾脏病理改变;蛋白质免疫印迹法检测大鼠肾组织内TGF-β1、Smad2、Smad3、p-Smad2、p-Smad3、Smad7的蛋白表达水平。结果各组大鼠经温肾健脾活血泄浊方干预后,与模型组相比,中药低、中、高剂量组大鼠尿液中TGF-β1、肌酐、24小时尿蛋白定量水平显著下降(P<0.05);血清中TGF-β1、胱抑素-C及肌酐定量显著降低,且中药低、中、高剂量组大鼠血清TGF-β1、胱抑素-C及肌酐定量水平均随着中药剂量增加而降低,具有负相关性,差异有统计学意义(P<0.05)。与模型组比较,中药低、中、高剂量组大鼠肾组织中TGF-β1、Smad2、Smad3蛋白定量显著降低,且随着中药剂量增加而降低,但差异无统计学意义;Smad7随中药剂量增加而升高,且差异有统计学意义(P<0.05),具有正相关性。苏木精—伊红染色结果示:中药各剂量组大鼠肾小管病变程度较模型组显著降低,纤维化面积较模型组明显减少;中药低、中、高剂量组较模型组大鼠肾组织结构紊乱程度降低,肾间质炎性细胞浸润减少,纤维组织减少;中药中、高剂Objective To investigate the effect of Professor Jin Hongyuan’s warming kidney and strengthening spleen and promoting blood circulation and eliminating turbidity decoction on transforming growth factor in kidney tissue of rats with chronic kidney disease(CKD)and to discuss the role of TGF-β1-Smad2/3 signaling pathway.Methods 56 rats were randomly divided into the blank group with 13 rats,and the remaining 43 rats were given adenine solution to prepare CKD rat model.After modeling identification,10 rats in the blank group were selected as the blank control group,and 40 rats with successful modeling were randomly divided into the model group,low,medium and high dose groups of traditional Chinese medicine with 10 rats in each group.The concentrations of the low,medium and high dose groups were administrated by gavage in the dose of 9.18 g/(kg•d),18.36 g/(kg•d)and 36.72 g/(kg•d),respectively.The rats in the blank control group and the model group were given equal volume of normal saline for 4 weeks.After intervention,the general condition of rats was observed.The quantitative levels of urine protein,creatinine,TGF-β1 and serum TGF-β1,cystatin C and creatinine in 24 hours were detected by enzyme-linked immunosorbent assay.The pathological changes of kidney were observed by hematoxylin-eosin and Masson staining.The protein expression levels of TGF-β1,Smad2,Smad3,p-Smad2,p-Smad3 and Smad7 in rat renal tissue were detected by Western blot.Results Compared with the model group,the levels of TGF-β1,creatinine and 24-hour urinary protein in urine of rats in the low,medium and high dose groups of traditional Chinese medicine were significantly decreased(P<0.05).The quantitative levels of TGF-β1,cystatin C and creatinine in serum were significantly decreased,and the quantitative levels of TGF-β1,cystatin C and creatinine in serum of rats in the high,medium and low dose groups of traditional Chinese medicine were decreased with the increase of dose,with a negative correlation,and the difference was statistically

关 键 词：慢性肾脏病 温肾健脾活血泄浊方 大鼠肾组织 肾功能 肾脏纤维化 转化生长因子-β1-Smad2/3 

分 类 号：R285.5[医药卫生—中药学]