BMP-7对TGF-β_1诱导人近端肾小管上皮细胞转分化及细胞外基质分泌的影响
|
摘要
目的:观察骨形态发生蛋白-7(BMP-7)对转化生长因子-β1(TGF-β1)诱导人近端肾小管上皮细胞(HK-2)向间充质细胞转分化(EMT)及细胞外基质(ECM)分泌的影响,探讨其逆转肾间质纤维化的可能机制。
方法:将HK-2细胞在体外与TGF-β1或TGF-β1+BMP-7共培养,分成对照组,不同浓度TGF-β1诱导组,TGF-β1不同时间诱导组,3ng/ml TGF-β1加不同浓度BMP-7组。相差显微镜观察细胞形态的改变;间接免疫荧光测定α-SMA、E-cadherin的表达; RT-PCR检测α-SMA、E-cadherin、ColⅠα1、FN、PAI-1 mRNA的表达;Western Blot检测α-SMA、E-cadherin蛋白的表达;ELISA方法测定细胞上清液ColⅠ、FN的表达。
结果:(1)随着TGF-β1浓度的增加及作用时间的延长,HK-2细胞形态逐渐发生改变,由立方形铺路石样转变为梭形长条样;在3ng/ml TGF-β1诱导48h后再加入200ng/ml BMP-7组中,可见到已转变为梭形长条样的细胞大部分逆转回原来的形态。(2)免疫荧光检测显示,正常HK-2细胞表达E-cadherin、而无α-SMA表达,经3ng/ml TGF-β1刺激48h后,细胞浆中α-SMA表达明显增强,细胞质膜E-cadherin表达显著减弱;去除TGF-β1,再加入200ng/ml BMP-7干预48h,能明显抑制α-SMA的表达、恢复E-cadherin的表达。(3)TGF-β1在10ng/ml内呈剂量依赖性上调α-SMA mRNA的表达。在3ng/ml TGF-β1诱导下,α-SMA mRNA表达随作用时间的延长而增加,E-cadherin mRNA表达随作用时间的延长而逐渐减少。在3ng/ml TGF-β1与BMP-7共同干预下,α-SMA mRNA及蛋白的表达随着BMP-7(100 ng/ml ~400 ng/ml )的浓度增加而减少,400 ng/ml BMP-7即可显著减少α-SMA mRNA及蛋白的表达(P<0.01)。在3ng/ml TGF-β1的持续刺激下,E-cadherin mRNA及蛋白的表达随着BMP-7的浓度增加而逐渐增加,400 ng/ml BMP-7即可显著增加E-cadherin mRNA及蛋白的表达(P<0.01)。(4)在3ng/ml TGF-β1诱导下,随刺激时间的延长,ColⅠα1、FN mRNA表达逐渐增加; BMP-7呈剂量依赖性抑制ColⅠ、FN的表达, 400 ng/ml BMP-7即可显著抑制ColⅠ、FN的表达(P<0.01)。(5)在3ng/ml TGF-β1诱导下,PAI-1 mRNA表达明显增加(P<0.01);BMP-7呈剂量依赖性抑制PAI-1 mRNA表达,400ng/ml BMP-7可明显减少PAI-1 mRNA表达(P<0.01)。
结论:(1)TGF-β1呈剂量和时间依赖性上调α-SMA的表达、抑制E-cadherin的表达,诱导HK-2细胞转分化,并促进ColⅠ、FN的合成(。2)BMP-7可呈剂量依赖性地抑制TGF-β1诱导下α-SMA mRNA和蛋白的表达,并通过再诱导E-cadherin mRNA和蛋白的表达抑制、逆转TGF-β1诱导的EMT。(3)BMP-7可呈剂量依赖性抑制TGF-β1诱导下HK-2细胞外基质成分ColⅠ、FN mRNA和蛋白的表达;抑制PAI-1 mRNA的表达,即BMP-7可抑制TGF-β1诱导下肾小管上皮细胞ECM的合成、促进ECM降解。
Objective: To investigate the effects of bone morphogenic protein(BMP)-7 on epithelial-to-mesenchymal transition(EMT) and the expression of fibronectin(FN) and type collagenⅠ(ColⅠ) in human renal proximal tubular cells(HK-2) induced by transforming growth factor-β1(TGF-β1),and to explore the possible mechanisms of BMP-7 for the inhibition and reversal of renal interstitial fibrosis.
Methods: HK-2 cells were treated with TGF-β1 or a combination of TGF-β1 and BMP-7. HK-2 cells divided into the following groups according to different factors: Control group, TGF-β1(different time and concentration) group, 3ng/ml TGF-β1+ BMP-7 (different concentration) group. Morphological changes were assessed by phase contrast microscopy.The expression ofα-SMA and E-cadherin were analysed by indirect immunofluorescence ,RT-PCR and Western Blot respectively.The mRNA levels of extracelluar matrix components(ColⅠα1 and FN) and plasminogen activator inhibitior(PAI)-1 were measured by RT-PCR..The secretion of ColⅠand FN protein was quantitated by ELISA.
Results: (1) TGF-β1 treatment of confluent HK-2 cells resulted in distinct morphological changes in a time and dose-dependent manner.The control cells displayed typical cobblestone morphology of epithelial cells. 3ng/ml TGF-β1 induced profound morphologic changes after 48h, with cells becoming elongated in shape, but addition of 200ng/ml BMP-7 for 48h restored the epithelial morphology of the HK-2 cells.(2)Indirect immunofluorescence staining showed that expression ofα-SMA could not be seen in control cells but upregulated and enhanced by 3ng/ml TGF-β1 after 48h.Incubation with 200ng/ml BMP-7 for 48h dramatically abrogated TGF-β1-inducedα-SMA expression. Treatment of 3ng/ml TGF-β1 resulted in distinct loss of Ecadherin staining in the plasma membrane of HK-2 cells but subsequent treatment with 200ng/ml BMP-7 largely restored the E-cadherin protein staining. (3)The expression ofα-SMA mRNA were upregulated by TGF-β1 in a dose-dependent manner maximally at the concentration of 10ng/ml TGF-β1. The 3ng/ml TGF-β1-induced levels ofα-SMA mRNA and protein were increased while E-cadherin decreased in a time-dependent manner. BMP-7 dramatically supressed TGF-β1-inducedα-SMA mRNA and protein expression in a dose-dependent manner. 400 ng/ml BMP-7 could significantly block the expression ofα-SMA mRNA and protein induced by 3ng/ml TGF-β1(P<0.01).Under the continuous influence of 3ng/ml TGF-β1,increasing doses of BMP-7(100～400ng/ml) substantially restored the expression of E-cadherin mRNA and protein (P<0.01, 400 ng/ml BMP-7+ TGF-β1VS. TGF-β1 alone). (4)3ng/ml TGF-β1 could increase the mRNA expression levels of FN and ColⅠα1 in a time-depedent manner.BMP-7 could inhibit the expression of ECM components(FN, ColⅠ) induced by TGF-β1 in a dose-depedent manner:400ng/ml BMP-7 could supress the expression of FN and ColⅠsignificantly(P<0.01). (5) 3ng/ml TGF-β1 could increase the expression of PAI-1 mRNA(P<0.01 VS. control).BMP-7 could inhibit the expression of PAI-1 mRNA induced by TGF-β1 in a dose-dependent manner.400ng/ml BMP-7 could inhibit the expression of PAI-1 mRNA significantly(P<0.01).
Conclusions: (1) TGF-β1 can induce EMT and produce ECM components in association with an inhibition of E-cadherin expression, increase ofα-SMA, ColⅠand FN expression and loss of epithelial cell morphology.(2)BMP-7 can block and reverse TGF-β1-induced EMT by supressing the expression ofα-SMA and restoring the expression of E-cadherin in a dose-depedent manner.(3)BMP-7 can prevent accumulation of ECM components and enhance degradation of ECM components induced by TGF-β1 through reducing the expression of ColⅠ,FN and PAI-1.
方法:将HK-2细胞在体外与TGF-β1或TGF-β1+BMP-7共培养,分成对照组,不同浓度TGF-β1诱导组,TGF-β1不同时间诱导组,3ng/ml TGF-β1加不同浓度BMP-7组。相差显微镜观察细胞形态的改变;间接免疫荧光测定α-SMA、E-cadherin的表达; RT-PCR检测α-SMA、E-cadherin、ColⅠα1、FN、PAI-1 mRNA的表达;Western Blot检测α-SMA、E-cadherin蛋白的表达;ELISA方法测定细胞上清液ColⅠ、FN的表达。
结果:(1)随着TGF-β1浓度的增加及作用时间的延长,HK-2细胞形态逐渐发生改变,由立方形铺路石样转变为梭形长条样;在3ng/ml TGF-β1诱导48h后再加入200ng/ml BMP-7组中,可见到已转变为梭形长条样的细胞大部分逆转回原来的形态。(2)免疫荧光检测显示,正常HK-2细胞表达E-cadherin、而无α-SMA表达,经3ng/ml TGF-β1刺激48h后,细胞浆中α-SMA表达明显增强,细胞质膜E-cadherin表达显著减弱;去除TGF-β1,再加入200ng/ml BMP-7干预48h,能明显抑制α-SMA的表达、恢复E-cadherin的表达。(3)TGF-β1在10ng/ml内呈剂量依赖性上调α-SMA mRNA的表达。在3ng/ml TGF-β1诱导下,α-SMA mRNA表达随作用时间的延长而增加,E-cadherin mRNA表达随作用时间的延长而逐渐减少。在3ng/ml TGF-β1与BMP-7共同干预下,α-SMA mRNA及蛋白的表达随着BMP-7(100 ng/ml ~400 ng/ml )的浓度增加而减少,400 ng/ml BMP-7即可显著减少α-SMA mRNA及蛋白的表达(P<0.01)。在3ng/ml TGF-β1的持续刺激下,E-cadherin mRNA及蛋白的表达随着BMP-7的浓度增加而逐渐增加,400 ng/ml BMP-7即可显著增加E-cadherin mRNA及蛋白的表达(P<0.01)。(4)在3ng/ml TGF-β1诱导下,随刺激时间的延长,ColⅠα1、FN mRNA表达逐渐增加; BMP-7呈剂量依赖性抑制ColⅠ、FN的表达, 400 ng/ml BMP-7即可显著抑制ColⅠ、FN的表达(P<0.01)。(5)在3ng/ml TGF-β1诱导下,PAI-1 mRNA表达明显增加(P<0.01);BMP-7呈剂量依赖性抑制PAI-1 mRNA表达,400ng/ml BMP-7可明显减少PAI-1 mRNA表达(P<0.01)。
结论:(1)TGF-β1呈剂量和时间依赖性上调α-SMA的表达、抑制E-cadherin的表达,诱导HK-2细胞转分化,并促进ColⅠ、FN的合成(。2)BMP-7可呈剂量依赖性地抑制TGF-β1诱导下α-SMA mRNA和蛋白的表达,并通过再诱导E-cadherin mRNA和蛋白的表达抑制、逆转TGF-β1诱导的EMT。(3)BMP-7可呈剂量依赖性抑制TGF-β1诱导下HK-2细胞外基质成分ColⅠ、FN mRNA和蛋白的表达;抑制PAI-1 mRNA的表达,即BMP-7可抑制TGF-β1诱导下肾小管上皮细胞ECM的合成、促进ECM降解。
Objective: To investigate the effects of bone morphogenic protein(BMP)-7 on epithelial-to-mesenchymal transition(EMT) and the expression of fibronectin(FN) and type collagenⅠ(ColⅠ) in human renal proximal tubular cells(HK-2) induced by transforming growth factor-β1(TGF-β1),and to explore the possible mechanisms of BMP-7 for the inhibition and reversal of renal interstitial fibrosis.
Methods: HK-2 cells were treated with TGF-β1 or a combination of TGF-β1 and BMP-7. HK-2 cells divided into the following groups according to different factors: Control group, TGF-β1(different time and concentration) group, 3ng/ml TGF-β1+ BMP-7 (different concentration) group. Morphological changes were assessed by phase contrast microscopy.The expression ofα-SMA and E-cadherin were analysed by indirect immunofluorescence ,RT-PCR and Western Blot respectively.The mRNA levels of extracelluar matrix components(ColⅠα1 and FN) and plasminogen activator inhibitior(PAI)-1 were measured by RT-PCR..The secretion of ColⅠand FN protein was quantitated by ELISA.
Results: (1) TGF-β1 treatment of confluent HK-2 cells resulted in distinct morphological changes in a time and dose-dependent manner.The control cells displayed typical cobblestone morphology of epithelial cells. 3ng/ml TGF-β1 induced profound morphologic changes after 48h, with cells becoming elongated in shape, but addition of 200ng/ml BMP-7 for 48h restored the epithelial morphology of the HK-2 cells.(2)Indirect immunofluorescence staining showed that expression ofα-SMA could not be seen in control cells but upregulated and enhanced by 3ng/ml TGF-β1 after 48h.Incubation with 200ng/ml BMP-7 for 48h dramatically abrogated TGF-β1-inducedα-SMA expression. Treatment of 3ng/ml TGF-β1 resulted in distinct loss of Ecadherin staining in the plasma membrane of HK-2 cells but subsequent treatment with 200ng/ml BMP-7 largely restored the E-cadherin protein staining. (3)The expression ofα-SMA mRNA were upregulated by TGF-β1 in a dose-dependent manner maximally at the concentration of 10ng/ml TGF-β1. The 3ng/ml TGF-β1-induced levels ofα-SMA mRNA and protein were increased while E-cadherin decreased in a time-dependent manner. BMP-7 dramatically supressed TGF-β1-inducedα-SMA mRNA and protein expression in a dose-dependent manner. 400 ng/ml BMP-7 could significantly block the expression ofα-SMA mRNA and protein induced by 3ng/ml TGF-β1(P<0.01).Under the continuous influence of 3ng/ml TGF-β1,increasing doses of BMP-7(100～400ng/ml) substantially restored the expression of E-cadherin mRNA and protein (P<0.01, 400 ng/ml BMP-7+ TGF-β1VS. TGF-β1 alone). (4)3ng/ml TGF-β1 could increase the mRNA expression levels of FN and ColⅠα1 in a time-depedent manner.BMP-7 could inhibit the expression of ECM components(FN, ColⅠ) induced by TGF-β1 in a dose-depedent manner:400ng/ml BMP-7 could supress the expression of FN and ColⅠsignificantly(P<0.01). (5) 3ng/ml TGF-β1 could increase the expression of PAI-1 mRNA(P<0.01 VS. control).BMP-7 could inhibit the expression of PAI-1 mRNA induced by TGF-β1 in a dose-dependent manner.400ng/ml BMP-7 could inhibit the expression of PAI-1 mRNA significantly(P<0.01).
Conclusions: (1) TGF-β1 can induce EMT and produce ECM components in association with an inhibition of E-cadherin expression, increase ofα-SMA, ColⅠand FN expression and loss of epithelial cell morphology.(2)BMP-7 can block and reverse TGF-β1-induced EMT by supressing the expression ofα-SMA and restoring the expression of E-cadherin in a dose-depedent manner.(3)BMP-7 can prevent accumulation of ECM components and enhance degradation of ECM components induced by TGF-β1 through reducing the expression of ColⅠ,FN and PAI-1.
引文
[1] Muller GA,Zeisberg M,Strutz F.The importance of tubulointerstitial damage in progressive renal diseases[J].Nephrol Dial Transplant,2000,15(suppl6)76-77.
[2] Eddy AA. Moleucular basis of renal fibrosis[J].Pediatr Nephrol,2000,15(3-4):290-301.
[3] Yang J,Liu Y.Dissection of Key events in tubular epithelial to myofibroblast transition and its implications in renal interstitial fibrosis[J].Am J Pathol,2001,159(4):1465-1475.
[4]Liu Y.Epithelial to mesenchymal transition in renal fibrogenesis:pathologic significance,molecular mechanism,and therapeutic intervention[J].J Am Soc Nephrol,2004,16(1):1-12.
[5] Schnaper HW,Kopp JB.Renal fibrosis[J].Front Biosci,2003,8:e68-e86.
[6] Han SY, Jee YH, Han KH, et al. An imbalance between matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase-2 contributes to the development of early diabetic nephropathy [J].Nephrol Dial Transplant (2006) 21: 2406–2416.
[7] Andrew L,David JA. TGF-β signaling and the fibrotic response[J].FASEB J,2004,18(7):816-827.
[8] Tian YC,Chen YC,Huang CC,et al.Leptospiral outer membrane protein induces extracellular matrix accumulation through a TGF-β1/smad-dependent pathway [J]. .J Am Soc Nephrol 17: 2792–2798, 2006.
[9] Hu MC,Wasserman D,Hartwig S,etal.p38MAPK acts in the BMP7-dependent stimulatory pathway during epithelial cell morphogenesis and is regulated by Smad1[J].J Biol Chem,2004,279(13):12051-12059.
[10] Zeisberg M, Hanai J, Sugimoto H, et al. BMP-7 counteracts TGF-beta-induced epithelial-to-mesenchymal transition and reverses chronic renal injury. NatMed , 2003 , 9 (7) : 964-968.
[11] Wang S, Chen Q, Simon TC, et al. Bone morphogenic protein -7 (BMP - 7) , a novel therapy for diabetic nephropathy. Kidney Int ,2003 , 63 (6) : 2037 - 49.
[12] Zeisberg M. Bone morphogenic protein -7 and the kidney:current concepts and open questions.Nephrol Dial Transplant,2006,21:568-573.
[13] 程叙扬,李晓玫,王海燕,等.肾小管上皮细胞损伤在慢性肾脏疾病进展中的作用. 国外医学·泌尿系统分册,2002 ,22(3) :152 - 154.
[14] Kalluri R,Neilson EG..Epithelial-mesenchymal transition and its implications for fibrosis[J].J Clin Invest,2003,112(12):1776-1784.
[15] Li JH,Zhu HJ,Huang XR,etal.Smad7 inhibits fibrotic effect of TGF- Beta on renal tubularepithelial cells by blocking Smad2 activation[J].J Am Soc Nephrol,2002,13(6):1464-1472.
[16] Iwano M, Plieth D, Danoff TM, et al. Evidence that fibroblasts derive from ep ithe Lium during tissue fibrosis. J Clin Invest, 2002,110: 341 - 350.
[17] Yang JW,Liu YH. Blockage of tubular epithelial to myofibroblast transition by hepatocyte growth factor prevents renal interstitial fibrosis. J Am Soc Nephrol, 2002,13: 96–107.
[18] Cheng S, Lovett DH. Gelatinase A (MMP - 2) is necessary and sufficient for renal tubular cell epithelial - mesenchymal transformation.Am J Pathol, 2003, 162: 1937 - 1949.
[19] 李玉林主编.分子病理学.北京:人民卫生出版社,2002,383.
[20] Ogata Y,IshidoYA S,Fukuzaki A,et al.Upregulated expression of transforming growth factor-beta,type IV collagen,and plasminogen activator inhibitor-1 mRNA are decreased after release of unilateral ureteral obstruction. J EXP Med, 2002, 197(3): 159-168.
[21]Okada H,Mukoyama M,Nagae T,et al.Reduction in connective tissue growth factor by antisense treatment ameliorates renal tubulointerstitial fibrosis[J].J Am Soc Nephrol,2005,16(1)133-134.
[22] Wendy C, Stephen, Josephine M, et al. Connective tissue growth factor plays an important role in advanced glycation and product-induced tubular epithelial-to-mesenchymal transition: implication for diabetic renal disease. J Am Soc Nephrol, 2006,17:2484-2494.
[23] McMorrow T, Gaffney MM, Slattery G,et al. Cyclosporine A induced epithelial–mesenchymal transition in human renal proximal tubular epithelial cells. Nephrol Dial Transplant (2005) 20: 2215–2225.
[24] Chen D , Zhao M ,Mund Y GR. Bone morphogenetic proteins [J ] . Growth Factors ,2004 ,22 (4) :233-241.
[25] Niskic L ,Martin PY. BMP-7 (Bone morphogenetic protein-7) : a future for chronic renal failiure [ J ] . Rev Med Suisse ,2005 ,1 (8) :568-573.
[26] Shimasaki S, Moore RK, Otsuka F,et al.The Bone Morphogenetic protein system in mammalian reproduction [J ].Endocrine Reviews,2004, 25(1):72–101.
[27] Zeisberg M,Shah AA, Kalluri R. Bone morphogenic protein - 7 induces mesenchymal to epithelial transition in adult renal fibroblasts and facilitates regeneration of injured kidney. J Biol Chem, 2005 , 280 (9) :8094 - 100.
[28] 陈楠,李娅,刘峰,等.骨成形蛋白-7对人肾小管上皮细胞增殖和转分化的影响.中华肾脏病杂志.2004,20(5):343-346.
[29] 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.中华医学杂志.2006,86(8)544-548.
[30] Izumi N,, Mizuguchi S, Inagaki Y,et al.BMP-7 opposes TGF-β1-mediated collagen induction in mouse pulmonary myofibroblasts through Id2. Am J Physiol Lung Cell Mol Physiol 290: L120–L126, 2006.
[31] Wang S, Chen Q, Simon TC, et al. Bone morphogenic protein -7 (BMP - 7) , a novel therapy for diabetic nephropathy. Kidney Int ,2003 , 63 (6) : 2037 - 49.
[32] Ishidaya S,Ogata Y,Fukuzaki A,etal.Plasminogen activator inhibitor-1 and tissue-type plasminogen activator are up-regulated during unilateral ureteral obstruction in adult rats.J Urol,2002,167(3),1503-1507.
[33] Ogata Y, Ishidaya S, Fukuzaki A,etal.Upregulated expression of transforming growth factor-beta,type Ⅳ collagen,and plasminogen activator inhibitor-1 mRNA are decreased after release of unilateral ureteral obstruction.J Exp Med,2002,197(3),159-168.
[34] Wang SN , Hirschberg R. BMP7 antagonizes TGF-β-dependent fibrogenesis in mesangial cells[J ] . Am J Phy RenPhy ,2003 ,284 (5) : F1006.
[1] Eddy AA.Moleucular basis of renal fibrosis.Pediatr Nephrol,2000,15:290-301.
[2] 张翥,黄颂敏,付平. TGF -β在慢性肾脏病发生发展中的作用和抑制TGF -β的应用前景[J ].中国中西医结合肾病杂志,2003,4(9):557-558.
[3] J yoji Y, Masanori K, Mit suru K, et al. Differential im-munoexpressions of cytoskeletons in renal epithelial andinterstitial cells in rat and canine fibrotic kidneys , and in kidney-related cell lines under fibrogenic stimuli [J ] . Exp Toxc Pathol , 2005 , 57 (2) : 135.
[4] Liu YH. Epithelial to mesenchymal transition in renal fibrogenesis:pathologic significance. molecular mechanism. and therapeutic intervention. J Am Soc Nephrol,2004.15:1-12.
[5] Bottinger EP,Bitzer M. TGF-signaling in renal disease. J Am Soc Nephro1,2002,l3:2600-26l0.
[6] Inman GJ,Nicolas EJ, Hill CS.Nucleocytoplasmic shutting of Smad2,3 and Smad4 permits sensing of TGF receptor activity. Mol Cell,2002,10:283-294.
[7] Hironoby T, Hironobu MD. Pathogenesis of fibrosis:role of TGF-β and CTGF. Current Opinion, 2002,14(6):681-685.
[8] 韩敏,徐钢,曾锐,等. β-连环素酪氨酸磷酸化调控E-钙黏蛋白表达参与转化生长因子β1诱导的人肾小管上皮细胞转分化. 中华肾脏病杂志,2006,22(8):494-498.
[9] Ogata Y,IshidoYA S,Fukuzaki A,et al.Upregulated expression of transforming growth factor-beta,type IV collagen,and plasminogen activator inhibitor-1 mRNA are decreased after release of unilateral ureteral obstruction. J EXP Med, 2002, 197(3): 159-168.
[10] Ling H ,Li X,Jha S,et al. Therapeutic role of TGF-β1-neutralizing antibody in mouse cyclosporin A nephropathy: morphologic improvement associated with functional preservation[J].J Am Soc Nephrol, 2003, 14(2): 377-388.
[11] Chen D , Zhao M ,Mund Y GR. Bone morphogenetic proteins [J ] . Growth Factors ,2004 ,22 (4) :233-241.
[12] Zeisberg M,Shah AA, Kalluri R. Bone morphogenic protein - 7 induces mesenchymal to epithelial transition in adult renal fibroblasts and facilitates regeneration of injured kidney. J Biol Chem, 2005 , 280 (9) :8094 - 100.
[13] Niskic L ,Martin PY. BMP-7 (Bone morphogenetic protein-7) : a future for chronic renal failiure [ J ] . Rev Med Suisse ,2005 ,1 (8) :568-573.
[14] Craig A H , Ezra W ,Peter C , et al. Modulation of activin and BMP signaling [ J ] . Mol Cell Endocrinol , 2004 , 225 (1) :19.
[15] Mmorrissey J, Hryska K, Guo G, et al. Bone morphogenetic protein-7 improves renal fibrosis and accelerates the return of renal function. J Am Soc Nephrol , 2002 , 13 (1): S14 - 21.
[16] Wang S, Chen Q, Simon TC, et al. Bone morphogenic protein -7 (BMP - 7) , a novel therapy for diabetic nephropathy. Kidney Int ,2003 , 63 (6) : 2037 - 49.
[17] Zeisberg M, Hanai J, Sugimoto H, et al. BMP-7 counteracts TGF -beta1 - induced epithelial - to - mesenchymal transition and reverses chronic renal injury. NatMed , 2003 , 9 (7) : 964 - 8.
[18] Wang SN , Hirschberg R. BMP-7 antagonizes TGF-β-dependent fibrogenesis in mesangial cells[J ] . Am J Phy RenPhy ,2003 ,284 (5) : F1006.
[19] Zeisberg M, Bottiglo C, Kumar N, et al. Bonemorphogenic protein -7 inhibits progression of chronic renal fibrosis associated with two genetic mouse models. Am J Physiol Renal Physiol , 2003 , 285 (6) : F1060 -7.
[20] Wang SN , Hirschberg R. Bone morphogenetic protein-7 signals opposing transforming growth factor beta in mesangial cells[J ] . Bio Chem ,2004 ,279 (22) :23200-23206.
[21] Wang SN ,Lapage J , Hirschberg R. Loss of tubular bone morphogenetic protein-7 in diabetic nephropathy [ J ] . J Am Soc Nephrol , 2001 ,12 (3) :2392.
[22] Selbi W , Motte C , Hascall V , et al. BMP-7 modulates hyaluronan-mediated proximal tubular cell monocyte interaction[J ] . J Am Soc Nephrol ,2004 , 15 (5) :1199.
[23] Kopp J ,Effery B. BMP-7 and the proximal tubule [J ] . Kidey Int ,2002 , 61 (1) :351.
[2] Eddy AA. Moleucular basis of renal fibrosis[J].Pediatr Nephrol,2000,15(3-4):290-301.
[3] Yang J,Liu Y.Dissection of Key events in tubular epithelial to myofibroblast transition and its implications in renal interstitial fibrosis[J].Am J Pathol,2001,159(4):1465-1475.
[4]Liu Y.Epithelial to mesenchymal transition in renal fibrogenesis:pathologic significance,molecular mechanism,and therapeutic intervention[J].J Am Soc Nephrol,2004,16(1):1-12.
[5] Schnaper HW,Kopp JB.Renal fibrosis[J].Front Biosci,2003,8:e68-e86.
[6] Han SY, Jee YH, Han KH, et al. An imbalance between matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase-2 contributes to the development of early diabetic nephropathy [J].Nephrol Dial Transplant (2006) 21: 2406–2416.
[7] Andrew L,David JA. TGF-β signaling and the fibrotic response[J].FASEB J,2004,18(7):816-827.
[8] Tian YC,Chen YC,Huang CC,et al.Leptospiral outer membrane protein induces extracellular matrix accumulation through a TGF-β1/smad-dependent pathway [J]. .J Am Soc Nephrol 17: 2792–2798, 2006.
[9] Hu MC,Wasserman D,Hartwig S,etal.p38MAPK acts in the BMP7-dependent stimulatory pathway during epithelial cell morphogenesis and is regulated by Smad1[J].J Biol Chem,2004,279(13):12051-12059.
[10] Zeisberg M, Hanai J, Sugimoto H, et al. BMP-7 counteracts TGF-beta-induced epithelial-to-mesenchymal transition and reverses chronic renal injury. NatMed , 2003 , 9 (7) : 964-968.
[11] Wang S, Chen Q, Simon TC, et al. Bone morphogenic protein -7 (BMP - 7) , a novel therapy for diabetic nephropathy. Kidney Int ,2003 , 63 (6) : 2037 - 49.
[12] Zeisberg M. Bone morphogenic protein -7 and the kidney:current concepts and open questions.Nephrol Dial Transplant,2006,21:568-573.
[13] 程叙扬,李晓玫,王海燕,等.肾小管上皮细胞损伤在慢性肾脏疾病进展中的作用. 国外医学·泌尿系统分册,2002 ,22(3) :152 - 154.
[14] Kalluri R,Neilson EG..Epithelial-mesenchymal transition and its implications for fibrosis[J].J Clin Invest,2003,112(12):1776-1784.
[15] Li JH,Zhu HJ,Huang XR,etal.Smad7 inhibits fibrotic effect of TGF- Beta on renal tubularepithelial cells by blocking Smad2 activation[J].J Am Soc Nephrol,2002,13(6):1464-1472.
[16] Iwano M, Plieth D, Danoff TM, et al. Evidence that fibroblasts derive from ep ithe Lium during tissue fibrosis. J Clin Invest, 2002,110: 341 - 350.
[17] Yang JW,Liu YH. Blockage of tubular epithelial to myofibroblast transition by hepatocyte growth factor prevents renal interstitial fibrosis. J Am Soc Nephrol, 2002,13: 96–107.
[18] Cheng S, Lovett DH. Gelatinase A (MMP - 2) is necessary and sufficient for renal tubular cell epithelial - mesenchymal transformation.Am J Pathol, 2003, 162: 1937 - 1949.
[19] 李玉林主编.分子病理学.北京:人民卫生出版社,2002,383.
[20] Ogata Y,IshidoYA S,Fukuzaki A,et al.Upregulated expression of transforming growth factor-beta,type IV collagen,and plasminogen activator inhibitor-1 mRNA are decreased after release of unilateral ureteral obstruction. J EXP Med, 2002, 197(3): 159-168.
[21]Okada H,Mukoyama M,Nagae T,et al.Reduction in connective tissue growth factor by antisense treatment ameliorates renal tubulointerstitial fibrosis[J].J Am Soc Nephrol,2005,16(1)133-134.
[22] Wendy C, Stephen, Josephine M, et al. Connective tissue growth factor plays an important role in advanced glycation and product-induced tubular epithelial-to-mesenchymal transition: implication for diabetic renal disease. J Am Soc Nephrol, 2006,17:2484-2494.
[23] McMorrow T, Gaffney MM, Slattery G,et al. Cyclosporine A induced epithelial–mesenchymal transition in human renal proximal tubular epithelial cells. Nephrol Dial Transplant (2005) 20: 2215–2225.
[24] Chen D , Zhao M ,Mund Y GR. Bone morphogenetic proteins [J ] . Growth Factors ,2004 ,22 (4) :233-241.
[25] Niskic L ,Martin PY. BMP-7 (Bone morphogenetic protein-7) : a future for chronic renal failiure [ J ] . Rev Med Suisse ,2005 ,1 (8) :568-573.
[26] Shimasaki S, Moore RK, Otsuka F,et al.The Bone Morphogenetic protein system in mammalian reproduction [J ].Endocrine Reviews,2004, 25(1):72–101.
[27] Zeisberg M,Shah AA, Kalluri R. Bone morphogenic protein - 7 induces mesenchymal to epithelial transition in adult renal fibroblasts and facilitates regeneration of injured kidney. J Biol Chem, 2005 , 280 (9) :8094 - 100.
[28] 陈楠,李娅,刘峰,等.骨成形蛋白-7对人肾小管上皮细胞增殖和转分化的影响.中华肾脏病杂志.2004,20(5):343-346.
[29] 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.中华医学杂志.2006,86(8)544-548.
[30] Izumi N,, Mizuguchi S, Inagaki Y,et al.BMP-7 opposes TGF-β1-mediated collagen induction in mouse pulmonary myofibroblasts through Id2. Am J Physiol Lung Cell Mol Physiol 290: L120–L126, 2006.
[31] Wang S, Chen Q, Simon TC, et al. Bone morphogenic protein -7 (BMP - 7) , a novel therapy for diabetic nephropathy. Kidney Int ,2003 , 63 (6) : 2037 - 49.
[32] Ishidaya S,Ogata Y,Fukuzaki A,etal.Plasminogen activator inhibitor-1 and tissue-type plasminogen activator are up-regulated during unilateral ureteral obstruction in adult rats.J Urol,2002,167(3),1503-1507.
[33] Ogata Y, Ishidaya S, Fukuzaki A,etal.Upregulated expression of transforming growth factor-beta,type Ⅳ collagen,and plasminogen activator inhibitor-1 mRNA are decreased after release of unilateral ureteral obstruction.J Exp Med,2002,197(3),159-168.
[34] Wang SN , Hirschberg R. BMP7 antagonizes TGF-β-dependent fibrogenesis in mesangial cells[J ] . Am J Phy RenPhy ,2003 ,284 (5) : F1006.
[1] Eddy AA.Moleucular basis of renal fibrosis.Pediatr Nephrol,2000,15:290-301.
[2] 张翥,黄颂敏,付平. TGF -β在慢性肾脏病发生发展中的作用和抑制TGF -β的应用前景[J ].中国中西医结合肾病杂志,2003,4(9):557-558.
[3] J yoji Y, Masanori K, Mit suru K, et al. Differential im-munoexpressions of cytoskeletons in renal epithelial andinterstitial cells in rat and canine fibrotic kidneys , and in kidney-related cell lines under fibrogenic stimuli [J ] . Exp Toxc Pathol , 2005 , 57 (2) : 135.
[4] Liu YH. Epithelial to mesenchymal transition in renal fibrogenesis:pathologic significance. molecular mechanism. and therapeutic intervention. J Am Soc Nephrol,2004.15:1-12.
[5] Bottinger EP,Bitzer M. TGF-signaling in renal disease. J Am Soc Nephro1,2002,l3:2600-26l0.
[6] Inman GJ,Nicolas EJ, Hill CS.Nucleocytoplasmic shutting of Smad2,3 and Smad4 permits sensing of TGF receptor activity. Mol Cell,2002,10:283-294.
[7] Hironoby T, Hironobu MD. Pathogenesis of fibrosis:role of TGF-β and CTGF. Current Opinion, 2002,14(6):681-685.
[8] 韩敏,徐钢,曾锐,等. β-连环素酪氨酸磷酸化调控E-钙黏蛋白表达参与转化生长因子β1诱导的人肾小管上皮细胞转分化. 中华肾脏病杂志,2006,22(8):494-498.
[9] Ogata Y,IshidoYA S,Fukuzaki A,et al.Upregulated expression of transforming growth factor-beta,type IV collagen,and plasminogen activator inhibitor-1 mRNA are decreased after release of unilateral ureteral obstruction. J EXP Med, 2002, 197(3): 159-168.
[10] Ling H ,Li X,Jha S,et al. Therapeutic role of TGF-β1-neutralizing antibody in mouse cyclosporin A nephropathy: morphologic improvement associated with functional preservation[J].J Am Soc Nephrol, 2003, 14(2): 377-388.
[11] Chen D , Zhao M ,Mund Y GR. Bone morphogenetic proteins [J ] . Growth Factors ,2004 ,22 (4) :233-241.
[12] Zeisberg M,Shah AA, Kalluri R. Bone morphogenic protein - 7 induces mesenchymal to epithelial transition in adult renal fibroblasts and facilitates regeneration of injured kidney. J Biol Chem, 2005 , 280 (9) :8094 - 100.
[13] Niskic L ,Martin PY. BMP-7 (Bone morphogenetic protein-7) : a future for chronic renal failiure [ J ] . Rev Med Suisse ,2005 ,1 (8) :568-573.
[14] Craig A H , Ezra W ,Peter C , et al. Modulation of activin and BMP signaling [ J ] . Mol Cell Endocrinol , 2004 , 225 (1) :19.
[15] Mmorrissey J, Hryska K, Guo G, et al. Bone morphogenetic protein-7 improves renal fibrosis and accelerates the return of renal function. J Am Soc Nephrol , 2002 , 13 (1): S14 - 21.
[16] Wang S, Chen Q, Simon TC, et al. Bone morphogenic protein -7 (BMP - 7) , a novel therapy for diabetic nephropathy. Kidney Int ,2003 , 63 (6) : 2037 - 49.
[17] Zeisberg M, Hanai J, Sugimoto H, et al. BMP-7 counteracts TGF -beta1 - induced epithelial - to - mesenchymal transition and reverses chronic renal injury. NatMed , 2003 , 9 (7) : 964 - 8.
[18] Wang SN , Hirschberg R. BMP-7 antagonizes TGF-β-dependent fibrogenesis in mesangial cells[J ] . Am J Phy RenPhy ,2003 ,284 (5) : F1006.
[19] Zeisberg M, Bottiglo C, Kumar N, et al. Bonemorphogenic protein -7 inhibits progression of chronic renal fibrosis associated with two genetic mouse models. Am J Physiol Renal Physiol , 2003 , 285 (6) : F1060 -7.
[20] Wang SN , Hirschberg R. Bone morphogenetic protein-7 signals opposing transforming growth factor beta in mesangial cells[J ] . Bio Chem ,2004 ,279 (22) :23200-23206.
[21] Wang SN ,Lapage J , Hirschberg R. Loss of tubular bone morphogenetic protein-7 in diabetic nephropathy [ J ] . J Am Soc Nephrol , 2001 ,12 (3) :2392.
[22] Selbi W , Motte C , Hascall V , et al. BMP-7 modulates hyaluronan-mediated proximal tubular cell monocyte interaction[J ] . J Am Soc Nephrol ,2004 , 15 (5) :1199.
[23] Kopp J ,Effery B. BMP-7 and the proximal tubule [J ] . Kidey Int ,2002 , 61 (1) :351.