桑叶生物碱抑制糖尿病肾病大鼠肾间质纤维化的研究  

Inhibition of renal interstitial fibrosis by mulberry leaf alkaloids in rats with diabetic nephropathy

作　　者：吴锋[1] 韦丽[1] Wu Feng;Wei Li(Department of Endocrinology,Municipal Peoples Hospital,Baoji 721000,China)

机构地区：[1]宝鸡市人民医院内分泌科,宝鸡721000

出　　处：《临床肾脏病杂志》2022年第8期666-673,共8页Journal Of Clinical Nephrology

摘　　要：目的研究桑叶生物碱能否抑制糖尿病肾病(diabetic nephropathy,DN)大鼠肾间质纤维化。方法50只SD大鼠采用高脂饮食+尾静脉注射链脲佐菌素(streptozotocin,STZ)方法建立DN大鼠模型,将建模成功的大鼠用随机数字表分为模型组、贝那普利组、桑叶生物碱低/中/高剂量组,每组10只。10只SD大鼠同时予以标准饮食+尾静脉注射等体积枸橼酸钠缓冲液,记为对照组。贝那普利组予以1.7 mg/kg贝那普利灌胃,桑叶生物碱低、中、高剂量组分别予以50、100、200 mg/kg桑叶生物碱灌胃,模型组和对照组均予以4 mL/kg生理盐水灌胃,每天1次,共给药12周。全自动生化分析仪检测尿微量白蛋白(microalbuminuria,mAlb)、血肌酐(serum creatinine,Scr)、血尿素氮(blood urea nitrogen,BUN)并计算内生肌酐清除率(endogenous creatinine clearance rate,Ccr);使用苏木素-伊红(hematoxylin-eosin,HE)染色法染色,观察各组大鼠肾组织病理改变;Masson染色观察各组肾组织胶原纤维沉积并计算肾间质纤维化相对面积;实时逆转录荧光定量聚合酶链反应检测各组肾组织转化生长因子-β1(transforming growth factor-β1,TGF-β1)、Smad2、Smad3、结缔组织生长因子(connective tissue growth factor,CTGF)、纤维连接蛋白(fibronectin,FN)、信使核糖核酸(messenger RNA,mRNA)表达;蛋白质印迹法检测各组肾组织TGF-β1、Smad2、Smad3、CTGF、FN蛋白表达及p-Smad2、p-Smad3水平。结果模型组肾小球体积增大、系膜基质增生、基底膜增厚,肾小管上皮细胞空泡样变,肾间质有大量炎症细胞浸润,Masson染色观察可见大量蓝灰色胶原纤维沉积;桑叶生物碱3剂量组和贝那普利组血管肾组织病变及纤维化情况均有不同程度改善;与对照组比,模型组、桑叶生物碱3剂量组和贝那普利组mAlb、Scr、BUN和肾间质纤维化相对面积、TGF-β1、Smad2、Smad3、CTGF、FN mRNA和蛋白的表达及p-Smad2、p-Smad3水平升高,Ccr降低(P<0.05);与模型Objective To explore whether or not mulberry leaf alkaloids can inhibit renal interstitial fibrosis in diabetic nephropathy(DN)rats.Methods DN rat model was established by a high-fat diet and a tail vein injection of streptozotocin(STZ)in 50 Sprague-Dawley(SD)rats.The successfully modeled rats were randomly divided into five groups of model,benazepril and mulberry leaf alkaloid low/medium/high dose(n=10 each).In control group,10 rats received a standard diet and a tail vein injection of equal volume of sodium citrate buffer simultaneously.Benazepril group received 1.7 mg/kg benazepril,mulberry leaf alkaloid low/medium/high-dose groups 50/100/200 mg/kg mulberry leaf alkaloid and model and control groups 4 ml/kg normal saline once daily for 12 weeks.Urinary microalbumin(mAlb),serum creatinine(Scr)and blood urea nitrogen(BUN)were measured by an automatic biochemical analyzer and endogenous creatinine clearance(Ccr)was calculated.Hematoxylin eosin(HE)staining was utilized for observing the pathological changes of renal tissue in each group;Masson staining for observing the deposition of collagen fibers in renal tissue and calculating the relative area of renal interstitial fibrosis.Reverse transcription-quantitative polymerase chain reaction(RT qPCR)was utilized for detecting the expressions of transforming growth factorβ1(TGF-β1),connective tissue growth factor(CTGF),Smad2,Smad3 and fibronectin(FN)messenger RNA(mRNA)in renal tissue of each group.TGF-β1,Smad2,Smad3,CTGF,FN protein expression and p-Smad2,p-Smad3 levels in renal tissue of each group was detected by Western blot.Results In model group,glomerulus volume expanded,mesangial matrix proliferated,basement membrane thickened,renal tubular epithelial cells vacuolized and numerous inflammatory cells infiltrated in renal interstitium.Masson staining showed that a large number of blue gray collagen fibers were deposited.The pathological changes and fibrosis of vascular and renal tissues improved in varying degrees in mulberry leaf alkaloid low/medium/high-dose

关 键 词：桑叶生物碱 糖尿病肾病 肾间质纤维化 

分 类 号：R285.5[医药卫生—中药学]