肾小管上皮细胞转分化与基质金属蛋白酶的相关性研究
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摘要
目的:观察TGF-β1在诱导大鼠肾小管上皮细胞系(NRK-52E)的上皮-间充质转分化过程中,对细胞基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)的影响,探讨MMP-2和MMP-9与肾小管上皮细胞转分化的相关性。
方法:选择E钙粘附蛋白为上皮细胞标志物,α平滑肌动蛋白(α-SMA)为间叶组织标志物,以低血清培养的NRK-52E做为对照组,低血清培养的NRK-52E细胞经TGF-β1(10ng/ml)诱导72小时为实验组,分别以免疫细胞化学染色和RT-PCR方法检测NRK-52E的转分化状态, RT-PCR法观察NRK-52E转分化后的细胞MMP-2与MMP-9的表达。
结果:RT-PCR和免疫细胞化学染色结果均显示,NRK-52E细胞经TGF-β1体外诱导72小时后,其上皮细胞标志物E钙粘附蛋白的mRNA和蛋白的表达与相应的对照组相比明显减弱,同时其间叶组织标志物α-SMA mRNA和蛋白表达与相应的对照组相比均增强,表明培养的NRK-52E经TGF-β1体外诱导72小时发生了上皮-间充质细胞的转分化;此时,转分化细胞的MMP-2和MMP-9 mRNA表达均明显高于对照组细胞,说明TGF-β1诱导NRK-52E发生转分化后可使细胞的MMP-2与MMP-9表达增多。
结论:TGF-β1成功的诱导大鼠肾小管上皮细胞(NRK-52E)发生转分化,使细胞MMP-2与MMP-9mRNA表达增多。
Renal interstitial fibrosis is the final stage of many kinds of renal diseases[1]. Renal interstitial fibrosis is characteristic of the accumulation of excessive extra cellular matrix and the destruction of renal tissue induced by the development of kinds of renal diseases. Now, it is accepted that the excessive extra cellular matrix is secreted by the Myofibroblast[4], the precise resource of the Myofibroblast has been always considered activation of interstitial fibroblasts. But recently it was suggested that there might be other sources in addition to activation of interstitial fibroblasts. Some scientists have proposed that epithelial to mesenchymal transition is a major resource of the Myofibroblast,and renal tubular epithelial to mesenchymal transition plays an important role in renal interstitial fibrosis. So more and more scientists pay close attention to its role in the renal fibrosis. The so-called renal tubular epithelial cells to mesenchymal cell transdifferentiation usually pointed tubular epithelial cells lost E-cadherin, a marker of endothelial cells and acquired a phenotype of mesenchymal cells, a-SMA[26]. EMT of renal tubular cells can be affected by many factors, among these factors, TGF-β1[27] was proved to be an important cytokine which can induce the EMT of renal tubular cells. So TGF-β1 is selected to induce renal tubular epitheial cell to mesenchymal transition.
For the past few years, researches about the EMT of renal tubular cells emerged. It was noted that the integrity of tubular basement membrane plays an important role in renal tubular epithelial cell transdifferentiation. Tubular basement membrane is mainly composed of typeⅣcollagen, fibronectin and laminin and other extracellular matrix. While matrix metalloproteinases (MMPs) can digest the basement membrane. Specifically, MMP-2 and MMP-9 can digest collagenⅣ[32], which is the major component of glomerular basement membrane and renal tubular basement membrane. So more and more scientists pay close attention to its role in the EMT of renal tubular cells.
Purpose: In this study, renal tubular epithelial cell line (NRK-52E) be selected as the object, TGF-β1 to be selected as induction factor, we designed the experiment on the basis of above to observe the influence of TGF-β1 on MMP-2 and MMP-9, when it inducing the EMT of rat renal tubular cell line (NRK-52E). Detect the relationship between MMP-2, MMP-9 and the EMT of renal tubular cells.
Method: Rat renal tubular cell line (NRK-52E) was cultured in vivo. Performed Immunocytochemistry and RT-PCR to detect and clarify the EMT of NRK-52E cells 72 hours after been stimulated by TGF-β1(10ng/ml), selected E-cadherin as a marker of endothelial cells,α- SMA as a phenotype of mesenchymal cells. Then we observed the expression of MMP-2 and MMP-9 in the transformed NRK-52E cells. The NRK-52E cells cultured in the low- blood serum media was used as control.
Result: Growth of the normal cultured renal tubular epithelial cell (NRK-52E) showed a confluent monolayer of cells with a classical epithelial polygon cobblestone-like morphology.NRK-52E were cultured for 72 hours in the presence or absence of TGF-β1,we observed that that in the experimental group and control group has an obvious change in cell morphology. Growth of the renal tubular epithelial cell of control group always showed a confluent monolayer of cells with a classical epithelial polygon cobblestone-like morphology, whether , growth of the renal tubular epithelial cell of experimental group showed scattered singly, showed morphological changes typical of EMT, from epithelial cobblestone to fibroblastic spindle-shaped morphology. Rat renal tubular cell line (NRK-52E) was cultured in vivo, Performed Immunocytochemistry to detect and clarify the EMT of NRK-52E cells 72 hours after been stimulated by TGF-β1(10ng/ml), selected as a marker of endothelial cells,α- SMA as a phenotype of mesenchymal cells. Result showed that the expression of E-cadherin were stronger in the cell membrane of control group than experimental group,but, the expression ofα- SMA were less in the cell cytoplasm of control group than experimental group. Immunocytochemical results suggest that cultured NRK-52E cells in the induction of TGF-β1 has undergone epithelial - mesenchymal cell transdifferentiation. To further verify the immunocytochemistry result, RT-PCR to detect and clarify the EMT of NRK-52E cells 72 hours after been stimulated by TGF-β1(10ng/ml), selected E-cadherin as a marker of endothelial cells,α- SMA as a phenotype of mesenchymal cells.With the mRNA expression of GAPDH as an internal of RT-PCR, to RT-PCR detect and clarify the EMT of NRK-52E cells 72 hours after been stimulated by TGF-β1(10ng/ml), selected E-cadherin as a marker of endothelial cells,α- SMA as a phenotype of mesenchymal cells. Result showed that the mRNA expression of GAPDH were similarly, the mRNA expression of E-cadherin were stronger in the cell membrane of control group than experimental group, but, the mRNA expression ofα- SMA were less in the cell cytoplasm of control group than experimental group. It further verify the immunocytochemistry result. Result testified that TGF-β1 has successfully induced renal tubular epitheial cell to mesenchymal transition.In the same time,we observed the mRNA expression of MMP-2 and MMP-9 in the transformed NRK-52E cells. With the mRNA expression of GAPDH as an internal of RT-PCR,and the mRNA expression of GAPDH were similarly, result showed that the mRNA expression of MMP-2 were less in the cell cytoplasm of control group than experimental group, the mRNA expression of MMP-9 were less in the cell cytoplasm of control group than experimental group, then ,it further supported Yang and his co-worker′s conclusion that transformed HK-2 cells secrete a lot of MMP-2 and MMP-9 in the earlier process of renal tubular cells transit into the mesenchymal cells, then they raised that transformed NRK-52E cells secreted large quantities of MMP-2, MMP-9 were associated with tubular basement membrane disintegration of the basement membrane, then reinforcement of the capability of migration and invasion in the cells. The cells migrate to the interstitial area, secrete extra cellular matrix and promote the development of fibrosis.
Conclusion: TGF-β1 can induce rat renal tubular cells develop EMT, and the expression of MMP-2 and MMP-9 were enhanced in the transformed cells.
方法:选择E钙粘附蛋白为上皮细胞标志物,α平滑肌动蛋白(α-SMA)为间叶组织标志物,以低血清培养的NRK-52E做为对照组,低血清培养的NRK-52E细胞经TGF-β1(10ng/ml)诱导72小时为实验组,分别以免疫细胞化学染色和RT-PCR方法检测NRK-52E的转分化状态, RT-PCR法观察NRK-52E转分化后的细胞MMP-2与MMP-9的表达。
结果:RT-PCR和免疫细胞化学染色结果均显示,NRK-52E细胞经TGF-β1体外诱导72小时后,其上皮细胞标志物E钙粘附蛋白的mRNA和蛋白的表达与相应的对照组相比明显减弱,同时其间叶组织标志物α-SMA mRNA和蛋白表达与相应的对照组相比均增强,表明培养的NRK-52E经TGF-β1体外诱导72小时发生了上皮-间充质细胞的转分化;此时,转分化细胞的MMP-2和MMP-9 mRNA表达均明显高于对照组细胞,说明TGF-β1诱导NRK-52E发生转分化后可使细胞的MMP-2与MMP-9表达增多。
结论:TGF-β1成功的诱导大鼠肾小管上皮细胞(NRK-52E)发生转分化,使细胞MMP-2与MMP-9mRNA表达增多。
Renal interstitial fibrosis is the final stage of many kinds of renal diseases[1]. Renal interstitial fibrosis is characteristic of the accumulation of excessive extra cellular matrix and the destruction of renal tissue induced by the development of kinds of renal diseases. Now, it is accepted that the excessive extra cellular matrix is secreted by the Myofibroblast[4], the precise resource of the Myofibroblast has been always considered activation of interstitial fibroblasts. But recently it was suggested that there might be other sources in addition to activation of interstitial fibroblasts. Some scientists have proposed that epithelial to mesenchymal transition is a major resource of the Myofibroblast,and renal tubular epithelial to mesenchymal transition plays an important role in renal interstitial fibrosis. So more and more scientists pay close attention to its role in the renal fibrosis. The so-called renal tubular epithelial cells to mesenchymal cell transdifferentiation usually pointed tubular epithelial cells lost E-cadherin, a marker of endothelial cells and acquired a phenotype of mesenchymal cells, a-SMA[26]. EMT of renal tubular cells can be affected by many factors, among these factors, TGF-β1[27] was proved to be an important cytokine which can induce the EMT of renal tubular cells. So TGF-β1 is selected to induce renal tubular epitheial cell to mesenchymal transition.
For the past few years, researches about the EMT of renal tubular cells emerged. It was noted that the integrity of tubular basement membrane plays an important role in renal tubular epithelial cell transdifferentiation. Tubular basement membrane is mainly composed of typeⅣcollagen, fibronectin and laminin and other extracellular matrix. While matrix metalloproteinases (MMPs) can digest the basement membrane. Specifically, MMP-2 and MMP-9 can digest collagenⅣ[32], which is the major component of glomerular basement membrane and renal tubular basement membrane. So more and more scientists pay close attention to its role in the EMT of renal tubular cells.
Purpose: In this study, renal tubular epithelial cell line (NRK-52E) be selected as the object, TGF-β1 to be selected as induction factor, we designed the experiment on the basis of above to observe the influence of TGF-β1 on MMP-2 and MMP-9, when it inducing the EMT of rat renal tubular cell line (NRK-52E). Detect the relationship between MMP-2, MMP-9 and the EMT of renal tubular cells.
Method: Rat renal tubular cell line (NRK-52E) was cultured in vivo. Performed Immunocytochemistry and RT-PCR to detect and clarify the EMT of NRK-52E cells 72 hours after been stimulated by TGF-β1(10ng/ml), selected E-cadherin as a marker of endothelial cells,α- SMA as a phenotype of mesenchymal cells. Then we observed the expression of MMP-2 and MMP-9 in the transformed NRK-52E cells. The NRK-52E cells cultured in the low- blood serum media was used as control.
Result: Growth of the normal cultured renal tubular epithelial cell (NRK-52E) showed a confluent monolayer of cells with a classical epithelial polygon cobblestone-like morphology.NRK-52E were cultured for 72 hours in the presence or absence of TGF-β1,we observed that that in the experimental group and control group has an obvious change in cell morphology. Growth of the renal tubular epithelial cell of control group always showed a confluent monolayer of cells with a classical epithelial polygon cobblestone-like morphology, whether , growth of the renal tubular epithelial cell of experimental group showed scattered singly, showed morphological changes typical of EMT, from epithelial cobblestone to fibroblastic spindle-shaped morphology. Rat renal tubular cell line (NRK-52E) was cultured in vivo, Performed Immunocytochemistry to detect and clarify the EMT of NRK-52E cells 72 hours after been stimulated by TGF-β1(10ng/ml), selected as a marker of endothelial cells,α- SMA as a phenotype of mesenchymal cells. Result showed that the expression of E-cadherin were stronger in the cell membrane of control group than experimental group,but, the expression ofα- SMA were less in the cell cytoplasm of control group than experimental group. Immunocytochemical results suggest that cultured NRK-52E cells in the induction of TGF-β1 has undergone epithelial - mesenchymal cell transdifferentiation. To further verify the immunocytochemistry result, RT-PCR to detect and clarify the EMT of NRK-52E cells 72 hours after been stimulated by TGF-β1(10ng/ml), selected E-cadherin as a marker of endothelial cells,α- SMA as a phenotype of mesenchymal cells.With the mRNA expression of GAPDH as an internal of RT-PCR, to RT-PCR detect and clarify the EMT of NRK-52E cells 72 hours after been stimulated by TGF-β1(10ng/ml), selected E-cadherin as a marker of endothelial cells,α- SMA as a phenotype of mesenchymal cells. Result showed that the mRNA expression of GAPDH were similarly, the mRNA expression of E-cadherin were stronger in the cell membrane of control group than experimental group, but, the mRNA expression ofα- SMA were less in the cell cytoplasm of control group than experimental group. It further verify the immunocytochemistry result. Result testified that TGF-β1 has successfully induced renal tubular epitheial cell to mesenchymal transition.In the same time,we observed the mRNA expression of MMP-2 and MMP-9 in the transformed NRK-52E cells. With the mRNA expression of GAPDH as an internal of RT-PCR,and the mRNA expression of GAPDH were similarly, result showed that the mRNA expression of MMP-2 were less in the cell cytoplasm of control group than experimental group, the mRNA expression of MMP-9 were less in the cell cytoplasm of control group than experimental group, then ,it further supported Yang and his co-worker′s conclusion that transformed HK-2 cells secrete a lot of MMP-2 and MMP-9 in the earlier process of renal tubular cells transit into the mesenchymal cells, then they raised that transformed NRK-52E cells secreted large quantities of MMP-2, MMP-9 were associated with tubular basement membrane disintegration of the basement membrane, then reinforcement of the capability of migration and invasion in the cells. The cells migrate to the interstitial area, secrete extra cellular matrix and promote the development of fibrosis.
Conclusion: TGF-β1 can induce rat renal tubular cells develop EMT, and the expression of MMP-2 and MMP-9 were enhanced in the transformed cells.
引文
[1] Nahas AM. Pathways to renal fibrosis.Exp Nephrol 1995,3(2):71-75.
[2] Guo G,Morrissey J,McCracken R,et al.Role of TNFR1 and TNFR2 receptors in tubulointerstitial fibrosis of obstructive nephropathy [J].Am J Physiol,1999,277(5 2):766-772.
[3] Chevalier RL. Pathogenesis of renal injury in obstructive uropathy. CurrOpin Pediatr, 2006, 18(2): 153-160.
[4] Badid C, Mounier N, Costa AM, et al. Histol Histopathol,2000;15(1);269-80
[5] Okada H,Danoff TM, Kalluri R, Neilson EG: Early role of FSP1 in epithelial-mesenchymal transformation[ J ]..Am J Physiol1997,42: F563–F574
[6] Frank Sruta and Gerhard A.Muller. Renal fibrosis and the origin of the renal fibroblast. Nephrol Dial Transplant 2006; 21: 3368-3370
[7] Qi W , Chen X, Poronnik Pet al. The renal cortical fibroblast in renal tubulointerstitial fibrosis[ J ]. Int J Biochem Cell Biol 2006;38:1-5
[8] Christensen EI, Verrousr PJ. Interstitial fibrosis: tubular hypothesis versus glomerular hypotheseis. Kidney Int. 2008; 74(10):1262-1269
[9] Strutz F, Zeisberg M, Ziyadeh FN et al. Role of basic fibroblast growth factor-2 in epithelial-mesenchymal transformation. Kidney Int, 2002,61(5):1714-1728
[10]李慧凛,周秋根,郑法雷.肾小管上皮细胞转分化的概与病理过程.世界医学杂志,2004,8(1):34-35.
[11]戴胜川,张玉梅细胞转分化的病理生理意义.生命科学, 2006,18(1):58-61
[12] Song K H, Ko S H, Ahn Y B, et al. In vitro Transdifferentiationof adult Pancre- Aticacinar cellsintoinsulin-expressingcells. Biochem Biophys Res Commun, 2004, 316(4): 1094-1100
[13]刘娜肾小管上皮细胞转分化在肾间质纤维化中的研究进展国际泌尿系统杂志2006,26(6):854-857
[14] Frid M G, Kale V A, Stenmark K R. Mature vascular endothelium can give rise tosmooth muscle cells via endothelial mesenchymal transdifferrentiation: in vitro analysis. Circ Res, 2002, 90(11): 1189-1196
[15]姚翠微陈孝文梁东肾小管上皮细胞表型转化与肾间质纤维化中国中西医结合肾病杂志,2004,3,(5):176-178
[16] LAN HY. Tubular epithelial- myofibroblast transdifferentiation Mechanisms in proximal tubule cells[J]. Curr Opin Nephrol Hypertens,2003, 12(1): 25- 29.
[17] Yang J , Liu Y. Dissection of key event s in tubular epithelial to myofibroblast tr -ansition and it simplications in renal interstitial fi-brosis[J]. Am J Pathol , 2001 ,159 (4) :1465-1475.
[18] Zeisberg M , Hanai J , Sugimoto H , et al . BMP-7 counteracts TGF-Beta-1 induce-d epithelial-to-mesenchymal transition and reverses chronic renal injury. Nat Med , 2003 , 9 (7) : 964-968.
[19] Sappino AP,Schurch W,Gabbiani G.Differentiation repertoire of fibroblastic cells: expression of cytokeletal proteins as marker of phenotypic modulations.Lab Invest,1990, 63(2):144-161.
[20]陆海英张悦肾小管上皮细胞转分化及逆转的分子机制国际病理科学与临床杂志2006, 26(1);63-66
[21] Hay ED. An overview of epithelio-mesenchymal transformation. Acta Anat 1995 ,154 :8 - 20.
[22] Hay ED , Zuk A. Transformations between epithelium and mesenchyme : normal , pathological , and experimentally induced[ J ]. Am J Kidney Disease , 1995 ,26 :678 -690.
[23]蒲蕾樊均明上皮细胞转分化在肾脏纤维化中的机制国外医学泌尿系统分册2003,23(4);442-445
[24] Li Y, Yang J , Dai C , et al . Role for integrin-linked kinase in medi-ating tubular epit helial to mesenchymal transition and renal intersti-tial fibrogenesis[ J ].J Clin Invest , 2003 , 112 (4) : 503-516
[25] ForinoM, Torregrossa R, CeolM, et al. TGF-beta1 induces epithelial-mesenchyma -l transition, but not myofibroblast transdifferentiationof human kidney tubularepithel -ial cells in primary culture[ J ]. Int J Exp Pathol, 2006, 87 (3) : 197 - 208.
[26] Fan JM ,Huang XR ,Ng YY,et al . Interleukin-1 induces tubular epithelial-myo -fibroblast transdifferentiation through a transforming growth factor-beta1–depen dent mechanism in vitro[ J ]. Am J Kidney Dis , 2001 , 37 (4) : 820-831.
[27]陈爽TGF- beta诱导的肾小管上皮细胞发生转分化状态研究2006
[28] Liu Y. Epit helial to mesenchymal transition in renal fibrogenesis :pathologic sig -nificance , molecular mechanism , and therapeutic intervention[ J ]. Am J Soc Nephro-l ,2004 , 15 (1) : 1212.
[29] Feng XH,Derynck R.Specificity and versatility in TGF-beta singnaling through Sm -ads.Annu Rev Cell Dev Biol.2005 Nov 10,21,659-693.
[30] Dai C, Yang J,Liu Y. Transforming growth factor-beta1 potentiates renal tubular epithelial cell death by a mechanism independent of Smad signaling[ J ]. J Biol Chem , 2003, 278 (14) : 12537-12545
[31] Behrens P,Rothe M,Florin A,et al. Invasive properties of serous human epithelial ovarian tumors are related to Ets-1,MMP-1 and MMP-9 Express-ion[ J ].Int J Mol Med, 2001,8(2),149-54.
[32]张红梅孙树勋基质金属蛋白酶、基质金属蛋白酶组织抑制剂与肾脏疾病华北煤炭医学报2003,5(5);576-578
[33]袁移安基质金属蛋白酶-2和基质金属蛋白酶-9在肾脏纤维化中的作用2003
[34] Zeisberg M, Bonner G, Maeshima Y, et al. Renal Fibrosis : Collagen Composition and Assembly Regulates Epithelial Mesenchymal Transdifferentiation[J]. Am J Pathol , 2001 ,159 :1313 - 1321.
[2] Guo G,Morrissey J,McCracken R,et al.Role of TNFR1 and TNFR2 receptors in tubulointerstitial fibrosis of obstructive nephropathy [J].Am J Physiol,1999,277(5 2):766-772.
[3] Chevalier RL. Pathogenesis of renal injury in obstructive uropathy. CurrOpin Pediatr, 2006, 18(2): 153-160.
[4] Badid C, Mounier N, Costa AM, et al. Histol Histopathol,2000;15(1);269-80
[5] Okada H,Danoff TM, Kalluri R, Neilson EG: Early role of FSP1 in epithelial-mesenchymal transformation[ J ]..Am J Physiol1997,42: F563–F574
[6] Frank Sruta and Gerhard A.Muller. Renal fibrosis and the origin of the renal fibroblast. Nephrol Dial Transplant 2006; 21: 3368-3370
[7] Qi W , Chen X, Poronnik Pet al. The renal cortical fibroblast in renal tubulointerstitial fibrosis[ J ]. Int J Biochem Cell Biol 2006;38:1-5
[8] Christensen EI, Verrousr PJ. Interstitial fibrosis: tubular hypothesis versus glomerular hypotheseis. Kidney Int. 2008; 74(10):1262-1269
[9] Strutz F, Zeisberg M, Ziyadeh FN et al. Role of basic fibroblast growth factor-2 in epithelial-mesenchymal transformation. Kidney Int, 2002,61(5):1714-1728
[10]李慧凛,周秋根,郑法雷.肾小管上皮细胞转分化的概与病理过程.世界医学杂志,2004,8(1):34-35.
[11]戴胜川,张玉梅细胞转分化的病理生理意义.生命科学, 2006,18(1):58-61
[12] Song K H, Ko S H, Ahn Y B, et al. In vitro Transdifferentiationof adult Pancre- Aticacinar cellsintoinsulin-expressingcells. Biochem Biophys Res Commun, 2004, 316(4): 1094-1100
[13]刘娜肾小管上皮细胞转分化在肾间质纤维化中的研究进展国际泌尿系统杂志2006,26(6):854-857
[14] Frid M G, Kale V A, Stenmark K R. Mature vascular endothelium can give rise tosmooth muscle cells via endothelial mesenchymal transdifferrentiation: in vitro analysis. Circ Res, 2002, 90(11): 1189-1196
[15]姚翠微陈孝文梁东肾小管上皮细胞表型转化与肾间质纤维化中国中西医结合肾病杂志,2004,3,(5):176-178
[16] LAN HY. Tubular epithelial- myofibroblast transdifferentiation Mechanisms in proximal tubule cells[J]. Curr Opin Nephrol Hypertens,2003, 12(1): 25- 29.
[17] Yang J , Liu Y. Dissection of key event s in tubular epithelial to myofibroblast tr -ansition and it simplications in renal interstitial fi-brosis[J]. Am J Pathol , 2001 ,159 (4) :1465-1475.
[18] Zeisberg M , Hanai J , Sugimoto H , et al . BMP-7 counteracts TGF-Beta-1 induce-d epithelial-to-mesenchymal transition and reverses chronic renal injury. Nat Med , 2003 , 9 (7) : 964-968.
[19] Sappino AP,Schurch W,Gabbiani G.Differentiation repertoire of fibroblastic cells: expression of cytokeletal proteins as marker of phenotypic modulations.Lab Invest,1990, 63(2):144-161.
[20]陆海英张悦肾小管上皮细胞转分化及逆转的分子机制国际病理科学与临床杂志2006, 26(1);63-66
[21] Hay ED. An overview of epithelio-mesenchymal transformation. Acta Anat 1995 ,154 :8 - 20.
[22] Hay ED , Zuk A. Transformations between epithelium and mesenchyme : normal , pathological , and experimentally induced[ J ]. Am J Kidney Disease , 1995 ,26 :678 -690.
[23]蒲蕾樊均明上皮细胞转分化在肾脏纤维化中的机制国外医学泌尿系统分册2003,23(4);442-445
[24] Li Y, Yang J , Dai C , et al . Role for integrin-linked kinase in medi-ating tubular epit helial to mesenchymal transition and renal intersti-tial fibrogenesis[ J ].J Clin Invest , 2003 , 112 (4) : 503-516
[25] ForinoM, Torregrossa R, CeolM, et al. TGF-beta1 induces epithelial-mesenchyma -l transition, but not myofibroblast transdifferentiationof human kidney tubularepithel -ial cells in primary culture[ J ]. Int J Exp Pathol, 2006, 87 (3) : 197 - 208.
[26] Fan JM ,Huang XR ,Ng YY,et al . Interleukin-1 induces tubular epithelial-myo -fibroblast transdifferentiation through a transforming growth factor-beta1–depen dent mechanism in vitro[ J ]. Am J Kidney Dis , 2001 , 37 (4) : 820-831.
[27]陈爽TGF- beta诱导的肾小管上皮细胞发生转分化状态研究2006
[28] Liu Y. Epit helial to mesenchymal transition in renal fibrogenesis :pathologic sig -nificance , molecular mechanism , and therapeutic intervention[ J ]. Am J Soc Nephro-l ,2004 , 15 (1) : 1212.
[29] Feng XH,Derynck R.Specificity and versatility in TGF-beta singnaling through Sm -ads.Annu Rev Cell Dev Biol.2005 Nov 10,21,659-693.
[30] Dai C, Yang J,Liu Y. Transforming growth factor-beta1 potentiates renal tubular epithelial cell death by a mechanism independent of Smad signaling[ J ]. J Biol Chem , 2003, 278 (14) : 12537-12545
[31] Behrens P,Rothe M,Florin A,et al. Invasive properties of serous human epithelial ovarian tumors are related to Ets-1,MMP-1 and MMP-9 Express-ion[ J ].Int J Mol Med, 2001,8(2),149-54.
[32]张红梅孙树勋基质金属蛋白酶、基质金属蛋白酶组织抑制剂与肾脏疾病华北煤炭医学报2003,5(5);576-578
[33]袁移安基质金属蛋白酶-2和基质金属蛋白酶-9在肾脏纤维化中的作用2003
[34] Zeisberg M, Bonner G, Maeshima Y, et al. Renal Fibrosis : Collagen Composition and Assembly Regulates Epithelial Mesenchymal Transdifferentiation[J]. Am J Pathol , 2001 ,159 :1313 - 1321.