自拟宝肾方对糖尿病大鼠肾组织CTGF和MMP-9的影响
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摘要
糖尿病肾病(Diabetic nephropathy,DN)是糖尿病严重的微血管并发症,是导致终末期肾病的重要原因。其发病机理尚未完全阐明,一般认为它与高血糖、多元醇-肌醇代谢异常、氧化应激、血流动力学障碍、凝血机制障碍、蛋白的非酶促糖基化等多种因素协同作用有关。糖尿病肾病重要的病理改变之一就是细胞外基质(Extracellular matrix,ECM)的堆积。转化生长因子-β_1(Transformin growthfactorβ_1,TGF-β_1)和结缔组织生长因子(Connective tissue growth factor,CTGF)目前己被公认为是包括糖尿病肾小球硬化在内的多种纤维化病变发病机制中的驱动因子,是抗纤维化治疗的有效靶点。研究证实,转化生长因子-β_1可调节炎症反应,促进细胞外基质的合成和沉积,减少细胞外基质的降解,其过度表达可引起肾小球硬化和肾间质纤维化。结缔组织生长因子(CTGF)是TGF-β_1的下游效应介质。研究发现CTGF不介导TGF-β_1的抗增殖和抗炎作用,仅介导TGF-β_1过度表达时促细胞外基质合成的作用。它可以通过自分泌与旁分泌的形式,刺激肾小球系膜细胞和其他肾脏固有细胞的增殖,促进细胞外基质的堆积,参与着肾小球肥大、肾小球硬化以及间质纤维化。
基质金属蛋白酶(Matrix metalloproteinase,MMPs)主要起到降解细胞外基质的作用。基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)又称明胶酶B,是MMPs家族的主要成员之一,其作用底物主要是Ⅳ、Ⅴ、Ⅶ、Ⅹ型胶原、明胶以及弹性纤维。MMP-9以酶原形式分泌,必须激活后才能降解胶原及其它基质蛋白。普遍认为是纤溶酶原在纤溶酶原激活物的作用下活化为纤溶酶,后者以级联放大的瀑布效应激活MMP-9而导致基质降解。
腹腔注射链脲佐菌素(Streptozotocin,STZ)可在短时间内建立糖尿病大鼠模型,并出现糖尿病肾脏损害。本文以此模型为基础,探讨糖尿病肾病早期结缔组织生长因子和基质金属蛋白酶-9的表达以及自拟中药宝肾方治疗糖尿病肾病的机制。
目的:本研究以自拟中药宝肾方汤剂干预实验性糖尿病大鼠,应用免疫组化方法检测对照组(C组)、糖尿病组(D组)、糖尿病宝肾方治疗组(DB组)肾组织CTGF和MMP-9表达水平,并检测各组大鼠的生化指标,探讨自拟中药宝肾方汤剂保护肾脏的作用机理,为临床防治糖尿病肾病提供实验性理论依据。
方法:将实验动物Wistar大鼠随机分为对照组(C组)、糖尿病组(D组)、糖尿病宝肾方治疗组(DB组)。对照组大鼠腹腔注射枸橼酸缓冲液(假造模),糖尿病组大鼠腹腔注射链脲佐菌素(STZ)建立糖尿病大鼠模型,糖尿病宝肾方治疗组是在建立糖尿病模型基础上给予宝肾方汤剂灌胃。在实验8周后,各组大鼠留取血、尿以及肾脏标本,观察三组大鼠肾重(Weight of kidney)、体重(Bodyweight)、24小时尿蛋白(24hUpro)、血糖(Glu)、胆固醇(CHO)、尿素氮(BUN)、血肌酐(Scr)以及肾组织病理学变化;并通过免疫组织化学方法检测各组大鼠肾组织中CTGF和MMP-9的表达情况。
结果:和对照组相比,糖尿病组肾组织CTGF蛋白表达明显升高[(0.234±0.009)vs(0.146±0.007),P＜0.01],而MMP-9蛋白表达明显降低[(0.146±0.006)vs(0.236±0.007),P＜0.01],肾脏肥大指数、24小时尿蛋白、血糖、胆固醇、尿素氮、血肌酐明显升高(P＜0.01);和糖尿病组相比,宝肾方治疗组肾组织CTGF蛋白表达降低[(0.194±0.008)vs(0.234±0.009),P＜0.01],而MMP-9蛋白表达升高[(0.175±0.007)vs(0.146±0.006),P＜0.01],肾脏肥大指数降低(P＜0.05),24小时尿蛋白、胆固醇、尿素氮、血肌酐也降低(P＜0.01)。
结论:自拟宝肾方对糖尿病大鼠肾脏具有保护作用,其机制可能与影响肾组织CTGF和MMP-9的表达有关。
Diabetic nephropathy is a serious cap vascular complication of diabetes.It is a very important cause bringing about the terminal stage of nephropathy.The pathogenesis of diabetic nephropathy is not clear.It is generally considered that it is interrelated with the synergy of Hyperglycemia,metabolic disturbance of polylol-phaseomannite,oxidizes stress,abnormality of hemodynamics,abnormality of zymoplastic mechanism,nonenzymatic promoted glycosylation.The accumulation of extracellular matrix is one of the important pathologic changes of diabetic nephropathy.Now,TGF-β_1 and CTGF are considered to be one of the important driver molecules for ECM accumulation in the pathogenesis of diverse forms renal fibrosis including glomerulosclerosis in diabetic nephropathy,and may constitute a more specific target for futuramic anti-fibrosis therapies.It is very important to keep the function of inflammatory reaction.It can influence the formation of extracellular matrix(ECM),which includes inducing synthesis of various matrix proteins,inhibiting expression of ECM degradation enzyme,disturbing synthesis and decomposition of ECM,and leading to excessive accumulation of ECM,which result in glomerulosclerosis at last.It was found that CTGF played a key role in the occurrence and development of diabetic nephropathy.CTGF is the downstream medium of transforming growth factor-β_1(TGF-β_1),but CTGF does not involve in the resisting proliferation and inflammation function of TGF-β_1,and only involve in accelerating cell matrix synthesis function of TGF-β_1.CTGF can induce mesangial cells and other renal resident cells producing ECM by autocrine and paracrine,thereafter accelerate the glomerular hypertrophy and glomerulosclerosis inevitably.
Matrix metalloproteinases are mainly play degradation of extracellular matrix. Matrix metalloproteinases-9 say again gelatinases B,which is one of the main members of the family of MMPs.Ⅳ,Ⅴ,Ⅶ,Ⅹtype collagen,gelatin and elastic fiber are the substrate.MMP-9 in the form of enzyme secretion,must be activated to degrade matrix collagen and other proteins.Generally considered fibrinolytic enzyme in fibrinolytic enzyme activator activation is under the action of fibrinolytic enzyme, the latter to cascade amplification waterfall effect MMP-9 to activate substrate biodegradation.
The intraperitoneal injection of streptozotocin was conducted to made models of diabetic rats within a short period of time and impairment of diabetic nephropathy occurred after that.This article aims at discuss the expressions of CTGF and MMP-9 at the early stage of diabetic nephropathy and the mechanism of the curative effects of self-made Bao'shen recipe on diabetic nephropathy.
Objective:The study use Bao'shen recipe to intervent diabetic rats and detect the expression level of CTGF and MMP-9 of renal tissue in control group,diabetic rat group and Bao'shen recipe treating diabetic rat group by immumohistochemistry way. In addition,in order to provide theoretical basis for the treatment of DN and the possible mechanism of protecting kidney,we have detected the clinical and metabolic parameters of this three groups.
Methods:Wistar rats were randomized divided into control group,diabetic rat group and Bao'shen recipe treatment group.The control group received intraperitoneal injection with citric acid buffer solution.STZ was injected through abdominal cavity to make models of diabetic rats.Bao'shen recipe treatment group was set up based on models of diabetic rats with feeding Bao'shen recipe.Blood, urine and renal tissue were obtained after 8 weeks.Blood glucose,cholesterol,urea nitrogen,serum creatinine,24 hour-urinary albumin,weight of kidney,body weight and the pathological changes of kidney tissues were measured.Technique of immunohistochemistry was also taken to measure the expression of CTGF and MMP-9 in this three groups.
Result:Compared with group C,expressions of CTGF were higher[(0.234±0.009) vs(0.146±0.007),P＜0.01],while the expressions of MMP-9 were lower[(0.146±0.006) vs(0.236±0.007),P＜0.01]in renal tissue of group D. Kidney hypertrophy index,24-hour urinary protein,blood glucose,cholesterol,urea nitrogen and serum creatinine increased significantly in the group D.The level of MMP-9 expression was advanced[(0.175±0.007) vs(0.146±0.006),P＜0.01],while the level of CTGF expression was reduced[(0.194±0.008) vs(0.234±0.009),P＜0.01] by using Bao'shen recipe.In group DB,kidney hypertrophy index,24-hour urinary protein,cholesterol,urea nitrogen and serum creatinine obviously lower than diabetic rat group.
Conclusion:Self-made Bao'shen recipe can depress the expressions of CTGF and increase the expressions of MMP-9 in the kidney of diabetic rats,as a result, which can be served as a protective effect to prevent progress of diabetic nephropathy.
基质金属蛋白酶(Matrix metalloproteinase,MMPs)主要起到降解细胞外基质的作用。基质金属蛋白酶-9(Matrix metalloproteinase-9,MMP-9)又称明胶酶B,是MMPs家族的主要成员之一,其作用底物主要是Ⅳ、Ⅴ、Ⅶ、Ⅹ型胶原、明胶以及弹性纤维。MMP-9以酶原形式分泌,必须激活后才能降解胶原及其它基质蛋白。普遍认为是纤溶酶原在纤溶酶原激活物的作用下活化为纤溶酶,后者以级联放大的瀑布效应激活MMP-9而导致基质降解。
腹腔注射链脲佐菌素(Streptozotocin,STZ)可在短时间内建立糖尿病大鼠模型,并出现糖尿病肾脏损害。本文以此模型为基础,探讨糖尿病肾病早期结缔组织生长因子和基质金属蛋白酶-9的表达以及自拟中药宝肾方治疗糖尿病肾病的机制。
目的:本研究以自拟中药宝肾方汤剂干预实验性糖尿病大鼠,应用免疫组化方法检测对照组(C组)、糖尿病组(D组)、糖尿病宝肾方治疗组(DB组)肾组织CTGF和MMP-9表达水平,并检测各组大鼠的生化指标,探讨自拟中药宝肾方汤剂保护肾脏的作用机理,为临床防治糖尿病肾病提供实验性理论依据。
方法:将实验动物Wistar大鼠随机分为对照组(C组)、糖尿病组(D组)、糖尿病宝肾方治疗组(DB组)。对照组大鼠腹腔注射枸橼酸缓冲液(假造模),糖尿病组大鼠腹腔注射链脲佐菌素(STZ)建立糖尿病大鼠模型,糖尿病宝肾方治疗组是在建立糖尿病模型基础上给予宝肾方汤剂灌胃。在实验8周后,各组大鼠留取血、尿以及肾脏标本,观察三组大鼠肾重(Weight of kidney)、体重(Bodyweight)、24小时尿蛋白(24hUpro)、血糖(Glu)、胆固醇(CHO)、尿素氮(BUN)、血肌酐(Scr)以及肾组织病理学变化;并通过免疫组织化学方法检测各组大鼠肾组织中CTGF和MMP-9的表达情况。
结果:和对照组相比,糖尿病组肾组织CTGF蛋白表达明显升高[(0.234±0.009)vs(0.146±0.007),P＜0.01],而MMP-9蛋白表达明显降低[(0.146±0.006)vs(0.236±0.007),P＜0.01],肾脏肥大指数、24小时尿蛋白、血糖、胆固醇、尿素氮、血肌酐明显升高(P＜0.01);和糖尿病组相比,宝肾方治疗组肾组织CTGF蛋白表达降低[(0.194±0.008)vs(0.234±0.009),P＜0.01],而MMP-9蛋白表达升高[(0.175±0.007)vs(0.146±0.006),P＜0.01],肾脏肥大指数降低(P＜0.05),24小时尿蛋白、胆固醇、尿素氮、血肌酐也降低(P＜0.01)。
结论:自拟宝肾方对糖尿病大鼠肾脏具有保护作用,其机制可能与影响肾组织CTGF和MMP-9的表达有关。
Diabetic nephropathy is a serious cap vascular complication of diabetes.It is a very important cause bringing about the terminal stage of nephropathy.The pathogenesis of diabetic nephropathy is not clear.It is generally considered that it is interrelated with the synergy of Hyperglycemia,metabolic disturbance of polylol-phaseomannite,oxidizes stress,abnormality of hemodynamics,abnormality of zymoplastic mechanism,nonenzymatic promoted glycosylation.The accumulation of extracellular matrix is one of the important pathologic changes of diabetic nephropathy.Now,TGF-β_1 and CTGF are considered to be one of the important driver molecules for ECM accumulation in the pathogenesis of diverse forms renal fibrosis including glomerulosclerosis in diabetic nephropathy,and may constitute a more specific target for futuramic anti-fibrosis therapies.It is very important to keep the function of inflammatory reaction.It can influence the formation of extracellular matrix(ECM),which includes inducing synthesis of various matrix proteins,inhibiting expression of ECM degradation enzyme,disturbing synthesis and decomposition of ECM,and leading to excessive accumulation of ECM,which result in glomerulosclerosis at last.It was found that CTGF played a key role in the occurrence and development of diabetic nephropathy.CTGF is the downstream medium of transforming growth factor-β_1(TGF-β_1),but CTGF does not involve in the resisting proliferation and inflammation function of TGF-β_1,and only involve in accelerating cell matrix synthesis function of TGF-β_1.CTGF can induce mesangial cells and other renal resident cells producing ECM by autocrine and paracrine,thereafter accelerate the glomerular hypertrophy and glomerulosclerosis inevitably.
Matrix metalloproteinases are mainly play degradation of extracellular matrix. Matrix metalloproteinases-9 say again gelatinases B,which is one of the main members of the family of MMPs.Ⅳ,Ⅴ,Ⅶ,Ⅹtype collagen,gelatin and elastic fiber are the substrate.MMP-9 in the form of enzyme secretion,must be activated to degrade matrix collagen and other proteins.Generally considered fibrinolytic enzyme in fibrinolytic enzyme activator activation is under the action of fibrinolytic enzyme, the latter to cascade amplification waterfall effect MMP-9 to activate substrate biodegradation.
The intraperitoneal injection of streptozotocin was conducted to made models of diabetic rats within a short period of time and impairment of diabetic nephropathy occurred after that.This article aims at discuss the expressions of CTGF and MMP-9 at the early stage of diabetic nephropathy and the mechanism of the curative effects of self-made Bao'shen recipe on diabetic nephropathy.
Objective:The study use Bao'shen recipe to intervent diabetic rats and detect the expression level of CTGF and MMP-9 of renal tissue in control group,diabetic rat group and Bao'shen recipe treating diabetic rat group by immumohistochemistry way. In addition,in order to provide theoretical basis for the treatment of DN and the possible mechanism of protecting kidney,we have detected the clinical and metabolic parameters of this three groups.
Methods:Wistar rats were randomized divided into control group,diabetic rat group and Bao'shen recipe treatment group.The control group received intraperitoneal injection with citric acid buffer solution.STZ was injected through abdominal cavity to make models of diabetic rats.Bao'shen recipe treatment group was set up based on models of diabetic rats with feeding Bao'shen recipe.Blood, urine and renal tissue were obtained after 8 weeks.Blood glucose,cholesterol,urea nitrogen,serum creatinine,24 hour-urinary albumin,weight of kidney,body weight and the pathological changes of kidney tissues were measured.Technique of immunohistochemistry was also taken to measure the expression of CTGF and MMP-9 in this three groups.
Result:Compared with group C,expressions of CTGF were higher[(0.234±0.009) vs(0.146±0.007),P＜0.01],while the expressions of MMP-9 were lower[(0.146±0.006) vs(0.236±0.007),P＜0.01]in renal tissue of group D. Kidney hypertrophy index,24-hour urinary protein,blood glucose,cholesterol,urea nitrogen and serum creatinine increased significantly in the group D.The level of MMP-9 expression was advanced[(0.175±0.007) vs(0.146±0.006),P＜0.01],while the level of CTGF expression was reduced[(0.194±0.008) vs(0.234±0.009),P＜0.01] by using Bao'shen recipe.In group DB,kidney hypertrophy index,24-hour urinary protein,cholesterol,urea nitrogen and serum creatinine obviously lower than diabetic rat group.
Conclusion:Self-made Bao'shen recipe can depress the expressions of CTGF and increase the expressions of MMP-9 in the kidney of diabetic rats,as a result, which can be served as a protective effect to prevent progress of diabetic nephropathy.
引文
[1].Collins A J, Kasiske B, Herzog C, et al.United States renal data system 2006 annual data report abstract.Am J Kidney Dis.2007, 49: 6-71.
[2].Agardh CD,Stenram U, Torffvit O,et al.Efects of inhibition of glycation and oxidative stress on the development of diabetic nephropathy in rats.J Diabetes Complications,2002,16(6):395-400.
[3].Mason RM, Wahab NA.Extracellular matrix metabolism in diabetic nephropathy.! Am Soc Nephrol,2003,14:1358-1373.
[4].Bradham DM,Igarashi A,PotterRL,et al.Connective tissue growth factona cysteine-rich mitogen secreted by human vascuclar endothelial cells is related to the SRC-induced immediate early gene product CEF-lO.Cell Biol,1991,H4(6):1285-1294.
[5].Bork P.The modular architecture of a new family of growth regulators related to connective tissue growth factor.FEBS Lett,1993,327(2):125-130.
[6].Grotendorst GR,Okochi H,Hayashi N.A novel transforming growth factor beta response element controls the expression of the connective tissue growth factor gene.Cell Growth Differ, 1996,7(4):469-480.
[7].Abdel-Wahab N,Weston BS,Roberts T,et al.Connective Tissue Growth Factor and Regulation of the Mesangial Cell Cycle:Role in Cellular Hypertrophy.J Am Soc Nephrol,2002,13(10):2437-2445.
[8].Liu BC,Chen Q, Luo DD,et al.Mechanisms of irbesartan in prevention of renal lesion in streptozotocin-induced diabetic rats.Acta Pharmacol Sin,2003,24:67-73.
[9].Weston BS,Wahab NA,Mason RM.CTGF mediates TGF-beta-induced fibronectin matrix deposition by upregulating active alpha5betal integrin in human mesangial cells .J Am Soc Nephrol,2003,14(3):601-610.
[10].Wang S,Denichilo M,Brubaker C,et al.Connective tissue growth factor in tubulointerstitial injury of diabetic nephropathy.Kidney Int,2001,60(1):96-105.
[11].Crean JK,Furlong F,Finlay D,et al.Connective tissue growth factor [CTGFJ/CCN2 stimulates mesangial cell migration through integrated dissolution of focal adhesion complexes and activation of cell polarization.FASEB J,2004,18(13):1541-1543.
[12].McLennan SV,Wang XY,Moreno V,et al.Connective tissue growth factor mediates high glucose effects on matrix degradation through tissue inhibitor of matrix metalloproteinase type 1:implications for diabetic nephropathy.Endocrinology,2004,145(12):5646-5655.
[13].亓文波,陈凌,王兰芳,等.黄芪对糖尿病鼠肾脏氧化糖基化影响的实验研究.泰山医学院学报,2001,22(3):190-192.
[14].牟娜,张庆怡,倪兆慧,等.黄芪对高糖作用下肾间质成纤维细胞表达HGF 的影响.中国中西医结合肾病杂志,2002.3(1):7-9.
[15].杨黎明,蒋志宏.维生素E和硒对糖尿病肾病及高血压肾损害时丙二醛及超氧化物歧化酶的作用观察.上海医学,2002,25(11):687-689.
[16].鲁瑾,邹大进,张家庆.黄芪预防肿瘤坏死因子-a所致胰岛素抵抗.中国中西医结合杂志,1999,17(7):420-422.
[17].刘志强,李全志,秦贵军.黄芪注射液对早期糖尿病肾病患者血小板功能和血浆内皮素的影响.中国中西医结合杂志.2001,21(4):274-276.
[18].邱世翠,李清波,刘金荣,等.黄芪对小鼠免疫功能的影响.时珍国医国药,2000,11(5):389.
[19].陈琼,周端.藏红花对冠心病心绞痛患者血流变学的作用.辽宁中医杂志,1997,24(8):372-373.
[20].李再新,黄福.资源植物藏红花研究进展.四川食品与发酵,2005.4:1-4.
[21].邹建华.番红花提取物对纤维蛋白溶酶的影响.国外医学中医中药分册.1994,16(5):47.
[22].宁英远,王俭勤,屈逐林.大黄素对人肾成纤维细胞增殖的影响.中国中西医结合杂志,2000,20(2):105-106.
[23].郭啸华,刘志红,戴春笋,等.大黄酸抑制TGF-β_1诱导的肾小管上皮细胞及细胞外基质产生.肾脏病与透析肾移植杂志,2001,10(2):101-105.
[24].刘志红,朱加明,黄海东,等.大黄酸对转化生长因子诱导的内皮细胞纤溶酶原激活物抑制物1表达的影响.中华肾脏病杂志,2002,18(5):337-341.
[25].Bolzan AD,Bianchi MS.Genotoxicity of streptozotocin.Mutat Res,2002,512(2-3):121-134.
[26].Pirson Y,Physiopathology of diabetic nephropathy.what we learn from transplantation.Nephrologie,1998;19:105.
[27].刘道芳.黄芪研究进展信息.中医药信息,1998,15(2):13.
[28].王法德,王立琴.参芪克糖灵治疗糖尿病的实验研究.山东中医杂志,1997,16(7):316-319.
[29].郭啸华,刘志红,彭艾等.大黄酸对2型糖尿病大鼠疗效观察.中华肾脏病杂志,2002,18(4):280-284.
[30].雷载权,张廷模主编.中华临床中药学[下卷]第一版.北京:人民卫生出版社.1994:317-324.
[31].宗颂梅,李宗友.藏红花的最新研究进展.国外医学中医中药分册,1997,19(5):20-21.
[32].李今玉,金松竹,卢波.藏红花的鉴定及临床应用.中国初级卫生保健,2007,21(6):71-72.
[33].绪广林,余书勤,龚祝南等.西红花对大鼠实验性高脂血症的影响及机制研究.中国中药杂志,2005,30(5):369-372.
[34].Eddy AA,Giachelli GM.Renal expression of genes that promotes interstitial inflammation and fibrosis in rats with protein overload proteinuria.Kidney Int,1995,47(6):1546.
[35].Adler SG,Kang S W,Feld S,et al.Can glomerular mRNAs in human type 1diabetes be used to predict transition fromnor moalbuminuria to microalbuminuria.Am J kidney Dis,2002,40(1):184-188.
[36].张春,朱忠华,邓安国.结缔组织生长因子对人肾小管上皮细胞转分化影响的研究.中华肾脏病杂志,2003,19(6):374-377.
[37].卓文磊,张璟.反义寡核苷酸抑制TGF-β_1诱导人肾小管上皮细胞表达CTGF.重庆医学,2004,33(5):731-734.
[38].Zhu QY,Jiang YH.Current research into interrelationship of matrix proteinases-9 and kidney disease.Clinical Medical Journal of China,2006,13(4):639-641.
[39].姚新明,叶山东.基质金属蛋白酶-9与糖尿病肾病关系的研究进展.中国临床保健杂志,2007,10(5):542-545.
[40].Mclennan SV,Yue DK,Turtle JR.Effect of glucose on matrix metalloproteinase activity in mesangial cells.Nephron,1998,79(3):293- 298.
[41].马洪波,张木勋.MMP-9在糖尿病肾病中的研究进展.国外医学·老年医学分册,2003,24(3):114-118.
[42].Mclennan SV,Kelly DJ,Cox AJ,et al.Decreased matrix degradation in diabetic nephropathy:effects of ACE inhibition on the expression and activities of matrix metalloproteinases.Diabetologia,2002,45(2):268- 275.
[43].王秀芬,李英,王保兴等.糖尿病大鼠肾组织基质金属蛋白酶-9的表达及缬沙坦肾脏保护机制探讨.中华肾脏病杂志,2004,20(3):163-165.
[44].王巍巍,陈以平.黄芪牛蒡子不同配伍对糖尿病大鼠肾组织结缔组织生长因子的影响.上海中医药杂志,2008,42(9):63-66.
[45].黄海长,闵亚丽,李惊子等.黄芪当归合剂及依那普利对结缔组织生长因子在肾间质纤维化中表达的比较研究.中华肾脏病杂志,2003,19(3):133-136.
[46].刘宽芝,高贤荣,马军力等.三七、黄芪对2型糖尿病大鼠肾脏MMP-9表达的影响.中成药,2008,30(6):920-922.
[47].邹建华翻译.番红花提取物对纤维蛋白溶酶的影响.国外医学中医中药分册,1994,16(5):47.
[48].何晓峰,刘蔚,刘晓城等.黄芪对左肾静脉狭窄大鼠肾组织TGF-β_1和PAI-1表达的影响.中国组织化学与细胞化学杂志,2008,17(3):247-251.
[49].郑龙轶,文秀英,许明望等.贞清方对2型糖尿病大鼠肾组织基质金属蛋白酶-9和基质金属蛋白酶组织抑制物-1表达的调节作用.中国中西医结合杂志,2006,26:72-76.
[50].胡仲仪,陈以平,沈玲妹,等.黄芪注射液治疗糖尿病肾病性水肿的疗效观察.中国中西医结合杂志,2000,20(8):618.
[51].童黄锦,许惠琴.黄芪保护糖尿病肾病作用机制的研究进展.黑龙江中医药,2007,6:57-59.
[52].凌学静,张海石,黄岩.西红花对小鼠免疫增加作用的研究.中国中医基础医学杂志,1998.4(12):28.
[53].朱加明,刘志红,李颖健,等.大黄酸对葡萄糖转蛋白1基因转染系膜细胞功能的影响.中华内科杂志,2001,40(8):537-542.
[2].Agardh CD,Stenram U, Torffvit O,et al.Efects of inhibition of glycation and oxidative stress on the development of diabetic nephropathy in rats.J Diabetes Complications,2002,16(6):395-400.
[3].Mason RM, Wahab NA.Extracellular matrix metabolism in diabetic nephropathy.! Am Soc Nephrol,2003,14:1358-1373.
[4].Bradham DM,Igarashi A,PotterRL,et al.Connective tissue growth factona cysteine-rich mitogen secreted by human vascuclar endothelial cells is related to the SRC-induced immediate early gene product CEF-lO.Cell Biol,1991,H4(6):1285-1294.
[5].Bork P.The modular architecture of a new family of growth regulators related to connective tissue growth factor.FEBS Lett,1993,327(2):125-130.
[6].Grotendorst GR,Okochi H,Hayashi N.A novel transforming growth factor beta response element controls the expression of the connective tissue growth factor gene.Cell Growth Differ, 1996,7(4):469-480.
[7].Abdel-Wahab N,Weston BS,Roberts T,et al.Connective Tissue Growth Factor and Regulation of the Mesangial Cell Cycle:Role in Cellular Hypertrophy.J Am Soc Nephrol,2002,13(10):2437-2445.
[8].Liu BC,Chen Q, Luo DD,et al.Mechanisms of irbesartan in prevention of renal lesion in streptozotocin-induced diabetic rats.Acta Pharmacol Sin,2003,24:67-73.
[9].Weston BS,Wahab NA,Mason RM.CTGF mediates TGF-beta-induced fibronectin matrix deposition by upregulating active alpha5betal integrin in human mesangial cells .J Am Soc Nephrol,2003,14(3):601-610.
[10].Wang S,Denichilo M,Brubaker C,et al.Connective tissue growth factor in tubulointerstitial injury of diabetic nephropathy.Kidney Int,2001,60(1):96-105.
[11].Crean JK,Furlong F,Finlay D,et al.Connective tissue growth factor [CTGFJ/CCN2 stimulates mesangial cell migration through integrated dissolution of focal adhesion complexes and activation of cell polarization.FASEB J,2004,18(13):1541-1543.
[12].McLennan SV,Wang XY,Moreno V,et al.Connective tissue growth factor mediates high glucose effects on matrix degradation through tissue inhibitor of matrix metalloproteinase type 1:implications for diabetic nephropathy.Endocrinology,2004,145(12):5646-5655.
[13].亓文波,陈凌,王兰芳,等.黄芪对糖尿病鼠肾脏氧化糖基化影响的实验研究.泰山医学院学报,2001,22(3):190-192.
[14].牟娜,张庆怡,倪兆慧,等.黄芪对高糖作用下肾间质成纤维细胞表达HGF 的影响.中国中西医结合肾病杂志,2002.3(1):7-9.
[15].杨黎明,蒋志宏.维生素E和硒对糖尿病肾病及高血压肾损害时丙二醛及超氧化物歧化酶的作用观察.上海医学,2002,25(11):687-689.
[16].鲁瑾,邹大进,张家庆.黄芪预防肿瘤坏死因子-a所致胰岛素抵抗.中国中西医结合杂志,1999,17(7):420-422.
[17].刘志强,李全志,秦贵军.黄芪注射液对早期糖尿病肾病患者血小板功能和血浆内皮素的影响.中国中西医结合杂志.2001,21(4):274-276.
[18].邱世翠,李清波,刘金荣,等.黄芪对小鼠免疫功能的影响.时珍国医国药,2000,11(5):389.
[19].陈琼,周端.藏红花对冠心病心绞痛患者血流变学的作用.辽宁中医杂志,1997,24(8):372-373.
[20].李再新,黄福.资源植物藏红花研究进展.四川食品与发酵,2005.4:1-4.
[21].邹建华.番红花提取物对纤维蛋白溶酶的影响.国外医学中医中药分册.1994,16(5):47.
[22].宁英远,王俭勤,屈逐林.大黄素对人肾成纤维细胞增殖的影响.中国中西医结合杂志,2000,20(2):105-106.
[23].郭啸华,刘志红,戴春笋,等.大黄酸抑制TGF-β_1诱导的肾小管上皮细胞及细胞外基质产生.肾脏病与透析肾移植杂志,2001,10(2):101-105.
[24].刘志红,朱加明,黄海东,等.大黄酸对转化生长因子诱导的内皮细胞纤溶酶原激活物抑制物1表达的影响.中华肾脏病杂志,2002,18(5):337-341.
[25].Bolzan AD,Bianchi MS.Genotoxicity of streptozotocin.Mutat Res,2002,512(2-3):121-134.
[26].Pirson Y,Physiopathology of diabetic nephropathy.what we learn from transplantation.Nephrologie,1998;19:105.
[27].刘道芳.黄芪研究进展信息.中医药信息,1998,15(2):13.
[28].王法德,王立琴.参芪克糖灵治疗糖尿病的实验研究.山东中医杂志,1997,16(7):316-319.
[29].郭啸华,刘志红,彭艾等.大黄酸对2型糖尿病大鼠疗效观察.中华肾脏病杂志,2002,18(4):280-284.
[30].雷载权,张廷模主编.中华临床中药学[下卷]第一版.北京:人民卫生出版社.1994:317-324.
[31].宗颂梅,李宗友.藏红花的最新研究进展.国外医学中医中药分册,1997,19(5):20-21.
[32].李今玉,金松竹,卢波.藏红花的鉴定及临床应用.中国初级卫生保健,2007,21(6):71-72.
[33].绪广林,余书勤,龚祝南等.西红花对大鼠实验性高脂血症的影响及机制研究.中国中药杂志,2005,30(5):369-372.
[34].Eddy AA,Giachelli GM.Renal expression of genes that promotes interstitial inflammation and fibrosis in rats with protein overload proteinuria.Kidney Int,1995,47(6):1546.
[35].Adler SG,Kang S W,Feld S,et al.Can glomerular mRNAs in human type 1diabetes be used to predict transition fromnor moalbuminuria to microalbuminuria.Am J kidney Dis,2002,40(1):184-188.
[36].张春,朱忠华,邓安国.结缔组织生长因子对人肾小管上皮细胞转分化影响的研究.中华肾脏病杂志,2003,19(6):374-377.
[37].卓文磊,张璟.反义寡核苷酸抑制TGF-β_1诱导人肾小管上皮细胞表达CTGF.重庆医学,2004,33(5):731-734.
[38].Zhu QY,Jiang YH.Current research into interrelationship of matrix proteinases-9 and kidney disease.Clinical Medical Journal of China,2006,13(4):639-641.
[39].姚新明,叶山东.基质金属蛋白酶-9与糖尿病肾病关系的研究进展.中国临床保健杂志,2007,10(5):542-545.
[40].Mclennan SV,Yue DK,Turtle JR.Effect of glucose on matrix metalloproteinase activity in mesangial cells.Nephron,1998,79(3):293- 298.
[41].马洪波,张木勋.MMP-9在糖尿病肾病中的研究进展.国外医学·老年医学分册,2003,24(3):114-118.
[42].Mclennan SV,Kelly DJ,Cox AJ,et al.Decreased matrix degradation in diabetic nephropathy:effects of ACE inhibition on the expression and activities of matrix metalloproteinases.Diabetologia,2002,45(2):268- 275.
[43].王秀芬,李英,王保兴等.糖尿病大鼠肾组织基质金属蛋白酶-9的表达及缬沙坦肾脏保护机制探讨.中华肾脏病杂志,2004,20(3):163-165.
[44].王巍巍,陈以平.黄芪牛蒡子不同配伍对糖尿病大鼠肾组织结缔组织生长因子的影响.上海中医药杂志,2008,42(9):63-66.
[45].黄海长,闵亚丽,李惊子等.黄芪当归合剂及依那普利对结缔组织生长因子在肾间质纤维化中表达的比较研究.中华肾脏病杂志,2003,19(3):133-136.
[46].刘宽芝,高贤荣,马军力等.三七、黄芪对2型糖尿病大鼠肾脏MMP-9表达的影响.中成药,2008,30(6):920-922.
[47].邹建华翻译.番红花提取物对纤维蛋白溶酶的影响.国外医学中医中药分册,1994,16(5):47.
[48].何晓峰,刘蔚,刘晓城等.黄芪对左肾静脉狭窄大鼠肾组织TGF-β_1和PAI-1表达的影响.中国组织化学与细胞化学杂志,2008,17(3):247-251.
[49].郑龙轶,文秀英,许明望等.贞清方对2型糖尿病大鼠肾组织基质金属蛋白酶-9和基质金属蛋白酶组织抑制物-1表达的调节作用.中国中西医结合杂志,2006,26:72-76.
[50].胡仲仪,陈以平,沈玲妹,等.黄芪注射液治疗糖尿病肾病性水肿的疗效观察.中国中西医结合杂志,2000,20(8):618.
[51].童黄锦,许惠琴.黄芪保护糖尿病肾病作用机制的研究进展.黑龙江中医药,2007,6:57-59.
[52].凌学静,张海石,黄岩.西红花对小鼠免疫增加作用的研究.中国中医基础医学杂志,1998.4(12):28.
[53].朱加明,刘志红,李颖健,等.大黄酸对葡萄糖转蛋白1基因转染系膜细胞功能的影响.中华内科杂志,2001,40(8):537-542.