摘要
[目的]探讨解聚复肾宁(JJFSN)对糖尿病大鼠肾脏转化生长因子-β1(TGF-β1)mRNA,Ⅳ型胶原蛋白及肝细胞生长因子(HGF)表达的作用及其肾保护作用的机理。
[方法] 75只健康雄性SD大鼠,8周龄,体重200-220g,自由摄水1周后,单次腹腔注射链尿佐菌素法建立糖尿病大鼠模型,随机分为模型组(D组)、解聚复肾宁治疗组(J组)、依那普利治疗组(Y组)、JJFSN+依那普利治疗组(J+Y组),另设正常对照组(N组),采取相应干预措施处理12周后,检测各组大鼠体重(BW),肾指数,肾小球平均面积(MGPA)、肾小球平均体积(MGV)、空腹血糖(FBG)、血肌酐(Scr)、尿素氮(BUN)、24h尿白蛋白排泄率(UAER)、24h尿β2微球蛋白(β2-MG)、肌酐清除率(Ccr),原位杂交法检测大鼠肾组织TGF-β1mRNA表达,免疫组化法检测Ⅳ型胶原蛋白、HGF的表达,透射电镜观察肾组织超微结构。
[结果]各治疗组一般情况、生化指标、肾指数、MGPA、MGV变化均优于模型组,同时TGF-β1mRNA、Ⅳ型胶原蛋白表达明显下调,肾组织HGF表达明显上调,肾组织病理变化减轻,其中以J+Y组改善最显著(P<0.05)。
[结论] JJFSN对DM大鼠肾脏形态和功能有明显保护作用,其机制可能与其抑制TGF-β1mRNA、Ⅳ型胶原蛋白的表达,上调肾脏HGF表达,延缓肾脏纤维化进程有关。
Objective: To explore the effect of JJFSN on the expression of TGF-β1mRNA,Ⅳcollagen protein and HGF in the kidney of diabetic rats, and mechanism of its renal protection.
Methods: 75 SD rats, 8 weeks old, 200-220g, got one week’s free diet. Then 60 rats were choosed out randomly to establish diabetic model by STZ injection. And the 54 diabetic rats were divided into four groups:diabetic model group(D group, n=14), JJFSN group(J group, n=13), enalapril group(Y group, n=13), JJFSN + enalapril group(J + Y group, n=14), and the normal control group(N group, n=13).After treatment with the above responding drugs for 12 weeks, then the BW, KW/BW, MGPA, MGV, FBG, Scr, BUN, UAER,β2-MG, Ccr, were measured by routine analyse methods, the expression of TGF-β1 mRNA was detected by in situ Hybridization, the expression of HGF was detected by immuno- histochemistry, and kidney ultrastructure by transmission electron microscopy.
Result: Compare with D group, the fasting blood glucose, kidney index, serum creatinine, UEAR, MGPA, MGV were decreased significantly, and pathological abnormality obviously improved, especially in J+Y group; expression of TGF-β1mRNA andⅣcollagen were down-regulated significantly in 3 treatment groups.
Conclusion: JJFSN have some effect of protecting kidney of diabetic rats, which the mechanisms might relate to decreasing the levels of TGF-β1 andⅣcollagen expressing, upregulating the HGF, and inhibiting kidney hypertrophy.
引文
[1]胡仁明.内分泌代谢病临床新技术[M].第1版.北京:人民军医出版社, 2002,423-480.
[2]茅彩萍.糖尿病肾病发病机制的研究进展[J].中国血液流变学杂志,2002,12 (2):148-52.
[3]林善锬.糖尿病肾病发病机制的研究进展[J].中华内科杂志,2001,40(11):782-783.
[4] Mizuno S,Matsumoto K,Kurosawa T,et a1.Reciprocal balance of hepatocyte growth factor and transforming growth factor-βl in renal fibrosis in mice[J].Kidney Int,2000,57(3):937-948.
[5] Wang S,DeNichilo M,Brabaker C,et al. Connective tissue growth factor in tubulointerstitial injury of diabetic nephropathy[J].Kidney Int,2001,60(1):96-105.
[6] Manson RM,wahab NA.Extracellular matrix metabolism in diabetic nephmpathy [J].J Am Soc Nephrol,2003,14(5):1358-1373.
[7] Strutz F,Zeisberg M,RemiehausenA,et al.TGF-beta l induces proliferation in human renal fibroblasts via induction of basic fibroblast growth factor(EGF-2)[J].Kidney Int,2001,59(2):579-592
[8] Matsumoto K, Nakamura T. Hepatocyte growth factor: renotropic role and potential therapeutics for renal diseases [J]. Kidney Int, 2001, 59(6):2023-38.
[9]焦颖华,曹文富,陈益山,等.解聚复肾宁对糖尿病大鼠肾脏的保护作用[J].中药新药与临床药理,2008,19(2):91-93
[10]李伟,张红,殷松楼,等.不同剂量链脲佐菌素诱导SD大鼠糖尿病肾病模型的研究[J].徐州医学院学报,2006,26(1):52-55
[11]王永炎主编,中医内科学[M].上海科学技术出版社.1997,p301.
[12]李彦,中医药治疗糖尿病肾病研究进展[J].实用中医内科杂志,2008,22(10):45-47
[13]吕仁和,糖尿病及其并发症中西医诊治学[M].北京:人民卫生出版社,1997:528
[14]张晓丽,叶志斌,等.丹参在防治老年糖尿病肾病中的价值[J].老年医学与保健2008,l4(5):307-310
[15] Guo BL, Daisuke K, Motohide I, et al.Peroxisome proliferator-activated receptor-gamma ligands inhibit TGF-β1-induced fibronectin expression in glomerular mesangial cells[J].Diabetes,2004 ,53(1):200-208.
[16] Isono, Motohide, Andras Mogyorosi, Dong Cheol Han, et al. Stimulation of TGF-βtypeⅡreceptor by high gluocse in mouse mesangial cells and in diabetic kidney [J] .Am. J. Physiol. Renal Physiol, 2000, 278(5) :F830.
[17]高燕风,谢璇,吴娟娟,等.糖尿病肾病患者血浆转化生长因子β1测定及临床意义[J].中国糖尿病杂志,2002;10(6):376-379
[18] Park IS, Kiyomoto H, Annoud SL, et al. Expression of transforming growth factorβand IV collagen in early streptozocin induced diabetes[J].Diabetes,1997,46:924.
[19] Sassy Prigent C,Heudes D,Mandet C,et al.Early glomerular macrophage recruitment in streptozotocin induced diabetic rats.Diabetes ,2000,49(3):466-475.
[20]罗勇.糖尿病肾病与Ⅳ型胶原的关系[J].医学理论与实践,2005,18(7):745-747
[21] Singh R,Song RH,Alavi N,et al.High glucose decreases matrix metalloproteinase-2 activity in rat mesangial cells via transforming growth factor-beta1[J].Exp Nephrol,2001,9(4):249-257.
[22] Matsumoto K,Nakamura T.Hepatocyte growth factor:renotropic role and potenial therapeutics for renal disease[J].Kindey Int, 2001, 59:2023-2038.
[23] Liu Y, Rajur K, Tolbert E, et al.Endogenous hepatocyte growth factor ameliorates chronic renal injury by activating matrix degradation pathways [J]. Kidney Int,2000,58(5):2028-2043
[24] Naoki Y, Tomokazu N, Ikue M et al. Hepatocyte growth factor protects functional and histological disorders of HgCl2-induced acute renal failure mice. Nephron, 2002, 90(10):195-205
[25] Cruzado J,Lioheras N,Torras J,et al.Retression of advanced diabetic nephropathy by hepatocyte growth factor gene therapy in rats[J].Diabetes,2004,53(4):1119 -1127.
[26] Mizuno S, Nakamura T. Suppressions of chronic glomerular injuries and TGF-β1 production by HGF in attenuation of murine diabetic nephropathy [J].Am J Physiol Renal Physiol, 2004, 286(1) :F134 -F143.
[27] Dai C,Li YJ,Yang JW,et al. Hepatocyte growth factor preserves beta cell mass and mitigates hyperglycemia in streptozotocin-induced diabetic mice [J].J Biol Chem,2003, 278(29):27080-27087
[28] Kagami S,Border WA,Miller DE,et al.AngiotensinⅡstimulates extrea cellular matrix protein synthesis through inducdon of transforming growth factor expresion in rat glomemlar mesaugial cells[J].J Clin Invest,1994,93(6):2431.
[29]程红,田雪飞,董鸿瑞等.波生坦及依那普利对糖尿病肾病模型干预的对比研究[J].中华肾脏病杂志,2004,20(3):154-158
[30]陈慎仁,许文灿,林丹等.依那普利防治糖尿病肾病的疗效初探[J].中国药房,2002,13(7):418-420
[31]严萍萍,张文芳,李勇,等.黄芪注射液对心力衰竭患者神经内分泌系统的影响[J] .中华实用中西医杂志,2004,4 (17):3188-3190.
[32]王竹,王娟,吴喜利等,糖尿病肾病验方用药规律分析[J].陕西中医学院学报,2007,30(1):14-16
[33]牟姗,王琴,施蓓莉,等.高糖对肾小管上皮细胞胶原分泌和PAI-1合成的影响以及黄芪调控作用[J].中国中西医结合肾病杂志,2009,10(1):11-14
[34]黄映红,刘晓城,黄征宇,等.姜黄素对大鼠糖尿病肾的实验研究[J].中国现代医学杂志2004,14(7):94-97
[35]宁勇,刘晓城,等.姜黄素对糖尿病大鼠肾脏保护作用机制研究[J].湖北中医学院学报,2004,6(1):10-11
[36]陈丽,刘晓城,宁勇,姜黄素对糖尿病大鼠肾脏病变的作用及对肾脏Smad7表达的影响[J].华中科技大学学报(医学版) 2004,33(4):438-441
[37]熊礼佳,刘慎微,姜黄素对2型糖尿病大鼠肾组织OPN/NF-κB途径的影响[J].现代医学2008,36(1):15-18
[38]付燕,胡本容,汤强,等.药根碱、小檗碱、黄连煎剂及模拟方对小鼠血糖的影响[J].中草药, 2005, 36(4):548-551.
[39]游甦,刘波,黄连素对糖尿病患者尿Ⅳ型胶原影响的临床观察[J].中国医院药学杂志,2007,27(6):793-795
[40]刘慰华,黄河清,邓艳辉,等.黄连素对糖尿病肾损伤大鼠肾功能、氧化应激、肾脏醛糖还原酶的影响[J].中国药理学通报2008,24(7):955-959
[41]刘圣,唐丽琴,陈礼明,等.复方黄连胶囊对糖尿病肾病大鼠肾组织TGF-β1与Ⅳ型胶原蛋白表达的影响[J].中国中药杂志2008,33(1):68-72
[42]赵春云,刘素钗,温凤娥,等.贝那普利联合葛根素、前列地尔治疗糖尿病肾病临床疗效观察[J].临床荟萃2008,23(15):1117-1118
[43]陆秀锋,陆静,刘乃丰,等.氨基胍与葛根素对糖尿病大鼠肾脏结缔组织生长因子表达的影响[J].东南大学学报:医学版,2005,24(1):1317-1319
[44]张爱军,郭宏伟.丹参川芎嗪注射液联合地尔硫卓治疗早期糖尿病肾病疗效分析[J].数理医药学杂志, 2008, (6):676-677
[45]胡波,李锋,王燕午,等.丹参对糖尿病肾病大鼠MMP-2,TIMP-1,TGF-βl和Ⅳ表达的影响[J].时珍国医国药,2008,19(12):3020-3022
[46]柳刚,关广聚,亓同钢,等.丹参对糖尿病大鼠肾脏的保护作用及其机制研究[J].中西医结合学报,2005;3(6):459-462
[47]郭维玲,川芎嗪注射液和黄芪注射液联合应用对早期糖尿病肾病尿蛋白排泄率、肾功能的影响[J].实用中西医结合临床,2008,8(5):56
[48]张小如,彭建韫,杨明正,吴凌慧。川芎嗪对早期糖尿病肾病患者血清转化生长因子-βl的影响。浙江中医药大学学报,2007,31(4):435-436
[49]郑鸿翱,陈少如,陈穗等.当归或三七对兔甘油致急性肾衰的保护作用及其机制研究[J].中国现代医学杂志,2003年,9(9):46-49
[50]李世云,李道宽,牛素蒲,等.黄芪当归合剂治疗糖尿病肾病30例疗效观察[J].新中医,2008,40(6):21-22
[51]张家庆,关于糖尿病并发症中西医结合治疗及研究[J].中国中西医结合杂志,1996,16(1):3-4
[52]刘长山,黄芩甙对糖尿病患者血细胞醛糖还原酶活性及肾小球滤过率的影响[J].中华医学会第六次全国内分泌学术会议汇编。2001:广州:299-300;
[53]胡蜀红,张木勋。黄芩苷对STZ糖尿病大鼠肾脏的保护及其与TGF-βl的关系[J].中国医院药学杂志2007,27(3):290-293
[1] Massague J,Chen YG.Controlling TGF-beta signaling [J].Genes Dev,2000,14(6):627-644.
[2] Kolavennu V,Zeng L,Peng H,et a1.Targeting of RhoA/ROCK signaling ameliorates progression of diabetic nephropathy independent of glucose control [J].Diabetes,2008,57(3):714-723.
[3] Ziyadeh FN,Hoffman BB,Han DC,et al.Long-term prevention of renal insufficiency,excess matrix gene expression,and glomerular mesangial matrix expansion by treatment with monoclonal antitransforming growth factor-beta antibody in db/db diabetic mice[J].Proc Natl Acad Sci USA,2000,97(14):8015-8020
[4] Guha M,Xu ZG,Tung D,et a1.Specific down-regulation of connective tissue growth factor attenuates progression of nephropathy in mouse models of type l and type 2 diabetes [J].FASEB J,2007,21(12):3355-3368
[5] Fujita H,Omori S,Ishikura K,et a1. ERK and p38 mediate high-glucose-induced hypertrophy and TGF-beta expression in renal tubular cells[J]. Am J Physiol Renal Physiol,2004,286:F120-F126
[6]王丽晖,吴广礼,张丽霞,等,氟伐他汀对高糖培养的大鼠肾小球系膜细胞外基质p38 MAPK表达的影响. [J]药学学报,2009,44(2):121-125
[7] Kagami S,Border WA,Miller DE,et a1.AngiotensinⅡStimulates extracellutar matrix protein synthesis through induction of transforming growth factor-beta expression in rat glomerular mesangial cells[J].J Clin Invest,1994,93(6):2421-2427.
[8] Wolf G,Mueller E,Stahl RAK,et a1.AngiotensinⅡinduced hypertrophy of cultured murine proximal tubular cells is mediated by endogenous transforming growth factor-beta.[J]. J Clin Invest.1993.92:l366-l372.
[9]王巍巍,张金元,程劲,等,丹参对马兜铃酸肾损害大鼠肾组织病理学改变及ACE、ACE2表达的影响,[J]中国中西医结合肾病杂志,2009,10(2)109-113
[10] Chen S,Evans T,Deng D,et al. Hyperhexose mia induced functional and structural changes in the kidneys:role of endothelins [J].Nephron,2002,90(1):86-94.
[11]李晓苗,李源,曹雪滨,等,肾宁胶囊对2型糖尿病大鼠肾脏的保护作用. [J]第四军医大学学报,2009,30(3):217-220.
[12]郝娅宁,龚勇,尹爱萍,等.肾清饮对肾间质纤维化大鼠Ⅰ型胶原及TGF-β1表达的影响. [J]中国中西医结合肾病杂志,2009,10(1):21-24.
[13] Krussel JS,Bchr B,MilkiA,et a1.Vascular endothelial growth factor VEGF mRNA,splice variants are differentially expressed in human blastocysts [J].Mol Hum,Reprod,2001,7(1):57-63.
[14] Cha DR,Kim NH,Jung GHH.Expression of vascular endothelial growth factor in response to high glucose in rat mesangial cells[J].J Endocinol,2000,165 (3):617-624.
[15] Koizumi T,Shiraishi T,Hagihara N,et a1.Expression of vascular endothelial growth factors and their receptors in and around intracranial arteriovenous malformations[J].Neurosurgery,2002,50 (1):117-126
[16] Flyvbjerg A, Dagns - Hansen F, De Vriese AS, et al. Amelioration of long - term renal changes in obese type 2 diabetic mice by a neutralizing vascular endothelial growth factor antibody[J].Diabetes, 2002 ,51 (10):3090 -3094.
[17] Cooper M E.Interaction of metabolic and homodynamic factors in mediating experimental diabetic nephropathy[J].Diabetologia,2001,44(11):1957-72
[18]史永红,王晨,赵松,等,氟伐他汀对糖基化终末产物诱导肾小管细胞转分化及ERK1/2信号通路的影响. [J]中国药理学通报2008;24(10):1356-60
[19]杜晓霞,李学旺,李航,等,晚期糖基化终产物诱导人肾小管上皮细胞表达结缔组织生长因子[J].中华肾脏病杂志,2006. 22(12),753-757
[20]孙辽,余学清,祝胜郎,等,AGEs对肾小管上皮细胞转分化、1型胶原合成及smad信号通路的影响. [J]中国病理生理杂志,2006,22(12):2429-2433
[21]李长天,陈雁飞,葛根素对糖尿病大鼠肾脏糖基化终产物和转化生长因子-β1表达的影响. [J]中国中医药信息杂志,2006,13(3)36-38
[22]许艳芳,吴异兰,万建新,等,BMP-7对TTGF-β诱导人肾小管上皮细胞外基质表达的影响. [J]中国药理学通报2008;24(11):1453-1457
[23]谢席胜,刘衡川,樊均明,等,人参皂甙Rbl对转化生长因子-β1诱导的肾小管上皮细胞p47phox的表达及细胞外基质的影响.[J]四川大学学报(医学版)2009,40(1):106- l10
[24]李娅,陈楠,俞海瑾,等.骨形成蛋白-7基因转染对人肾小管上皮细胞外基质分泌的影响[J].中华医学杂志,2006,86(8):544-8.
[25]贾凤新,李亚秋,李桂,等,芪丹益肾降糖胶囊对糖尿病肾病患者T细胞亚群及血液TGF-β1含量变化的干预研究.[J]中华中医药学刊,2008,26(10):2201-2203
[26] Wang SN,Lapage J,Hirschberg R,et a1.Role of glomerular ultrafiltration of growth factors in progressive interstitial fibrosis in diabetic nephropathy.Kid hat,2000,57(3):l002-l0l4
