基于TGF-β1/Smad3信号通路研究肾炎1号方对单侧输尿管梗阻大鼠肾纤维化的改善作用  

Effects of Shenyan 1 Prescription on renal fibrosis improvement in rats with unilateral ureteral obstruction based on TGF-β1/Smad3 signaling pathway

作　　者：梁国强[1] 徐进 周丽霞 倪道磊[2] 任燕[2] 蒋春波[2] Liang Guoqiang;Xu Jin;Zhou Lixia;Ni Daolei;Ren Yan;Jiang Chunbo(Central Laboratory,Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,China;Nephrology Department,Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,China)

机构地区：[1]南京中医药大学附属苏州市中医医院中心实验室,苏州215009 [2]南京中医药大学附属苏州市中医医院肾内科,苏州1215009

出　　处：《国际中医中药杂志》2024年第1期42-48,共7页International Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82074368);江苏省高层次卫生人才科研项目(LGY2020046);江苏省中医药科技发展专项(2020ZX13);苏州市第五批和第九批姑苏卫生人才项目(GSWS2019063,GSWS2022107);苏州市科技计划项目(SKY2021058)。

摘　　要：目的研究肾炎1号方通过TGF-β1/Smad同源物3(Smad3)通路调控铁死亡对单侧输尿管梗阻(UUO)大鼠肾纤维化的保护作用及相关机制。方法将48只雄性SD大鼠按随机数字表法分为假手术组、模型组、肾炎1号方低(10 g生药/kg)、高(20 g生药/kg)剂量组,每组12只。除假手术组外,其余各组大鼠均采用梗阻单侧输尿管的方法复制UUO大鼠模型。造模完成后,各组灌胃相应药物或双蒸水,1次/d,连续4周。4周后,称重并测量左肾重量,采用生化分析法测定大鼠24 h尿蛋白定量和血清ALB、ALT、SCr、BUN水平;采用化学荧光测定试剂盒测定肾组织活性氧(ROS)水平,采用ELISA法测定大鼠左肾组织SOD、MDA水平;通过HE和普鲁士蓝染色法分别观察肾组织形态和含铁血黄素特异性蓝染情况;Western blot检测肾组织中TGF-β1、Smad3、谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族1成员5(SLC1A5)表达。结果与模型组比较,肾炎1号方高剂量组24 h尿蛋白定量降低(P<0.05),血清ALB水平升高(P<0.05),ALT水平降低(P<0.05),肾组织SLC1A5表达降低(P<0.05);肾炎1号方低、高剂量组左肾重量/体重降低(P<0.05);血清ROS、MDA水平降低(P<0.05),SOD活性明显升高(P<0.05);肾组织TGF-β1、Smad3表达降低(P<0.05),GPX4表达升高(P<0.05),并能改善肾组织病理损伤及铁离子沉积。结论肾炎1号方可能通过TGF-β1/Smad3通路调控铁死亡而抑制UUO大鼠肾纤维化,从而保护肾组织结构和功能。Objective To investigate the protective effects and mechanism of Shenyan 1 Prescription on renal fibrosis of unilateral ureteral obstruction(UUO)rats through TGF-β1/Smad homologous 3(Smad3)pathway regulating ferroptosis.MethodsTTotally 48 male SD rats were divided into four groups:sham-operation group,Uuo model group,and Shenyan 1 Prescription low-(10 drug/kg),and high-dosage(20 crude drug/kg)groups according to random number table method,with 12 rats in each group.The UUO model was induced by the method of unilateral ureteral obstruction except for those sham-operation group.After modeling,rats received corresponding drugs or normal saline by gavage for 4 weeks,once per day.After 4 weeks,the body mass and the left kidney weight were measured.The 24 h urine protein and the levels of serum albumin(ALB),alanine aminotransferase(ALT),serum creatinine(SCr)and blood urea nitrogen(BUN)were detected by biochemical analysis method;the ROs level in renal tissue was measured using a chemical fluorescence assay kit,and the SOD and MDA levels in left renal tissue of rats were measured using ELISA method;the morphology of renal tissue and the specific blue staining of hemosiderin were observed using HE and Prussian blue staining methods,respectively;the expressions of transforming growth factor-β1(TGF-β1),Smad3,glutathione peroxidase 4(GPX4),and solute carrier family 1 member 5(SLC1A5)were detected by Western blot.Results Compared with the model group,the 24 h urinary protein excretion in Shenyan 1 Prescription high-dosage group decreased(P<0.05),the serum ALB level increased(P<0.05),the ALT level decreased(P<0.05),and the expression of SLC1A5 in renal tissue decreased(P<0.05);the left kidney weight/body decreased in Shenyan 1 Prescription low-and high-dosage groups(P<0.05);the levels of serum ROS and MDA decreased(P<0.05),and the activity of SOD significantly increased(P<0.05);the expressions of TGF-β1 and Smad3 in renal tissue decreased(P<0.05),and the expression of GPX4 increased(P<0.05),and the renal pathological inju

关 键 词：铁死亡 肾纤维化 肾炎1号方 转化生长因子Β1 SMAD3蛋白 大鼠 

分 类 号：R285.5[医药卫生—中药学]