缬沙坦抑制人类肾小管上皮细胞转分化的初步研究
作者：文晓彦 郑法雷 孙阳 李艳 张晓明
单位：100730北京，中国医学科学院 中国协和医科大学 北京协和医院肾内科
关键词：肾小管上皮细胞；转分化；肌成纤维细胞；缬沙坦
中华肾病杂志000410摘 要：目的 探讨血管紧张素ⅡⅠ型受体拮抗剂缬沙坦(valsartan，Val)在人类肾小管上皮细胞系(HKC)转分化中的作用。方法 将培养的HKC细胞分为(1)无血清培养对照组；(2)阳性对照组(MCP-1+AAI组)：培养液中加人马兜铃酸-I(AAI)和单核细胞趋化蛋白-1(MCP-1)；(3)Val组：培养液中加入Val；(4)MCP-1+AAI+Val组。应用间接酶标免疫组织化学方法(IEI)检测HKC细胞。Α-平滑肌肌动蛋白(α-SMA)、波形蛋白、角蛋白表达的变化，流式细胞技术检测α-SMA阳性的HKC细胞百分数。结果 不同浓度缬沙坦(0.1、1.0、10.0、100.0pg/ml)分别作用于HKC细胞48h后，IEI测定细胞角蛋白、波形蛋白及α-SMA抗原表达，与无血清对照组比无明显变化；流式细胞术测得Val组表达α-SMA阳性的HKC细胞百分数分别为10.2%、5.6%、5.2%、0.9%，与无血清对照组(平均为3.1%)比较，差异无显著性意义(P>0.05)。MCP-1+AAI+Val(0.1、1.0、10.0pg/ml)组HKC细胞培养48h后，表达α-SMA阳性的HKC细胞分别为17.3%、32.4%、10.0%，与阳性对照组(平均为91.8%)比较，下降非常显著(P均<0.005)。结论 (1)缬沙坦单独作用对正常HKC细胞的转分化无明显影响。(2)缬沙坦对某些因素诱导的HKC细胞转分化可能具有显著抑制作用。
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Inhibition of transdifferentiation of human renal tubular epithelial cells by vaisartan
WEN Xiaoyan ,ZHENG Falei ,SUN Yang
(Division of Nephrology, Peking Union Medical College Hospital,Beijing 100730, China)
Abstract：Objective To investigate the possible role of valsartan(Val) in transdifferentiation of human renal tubular epithelial cell line(HKC).Methods HKC cells were divided into four groups:(1)serum-free(negative control);(2) MCP-1+AAI-treated (positive control);(3) Val-treated alone;(4)Val-inhibition(treated with MCP-1+ AAI+ Val).Then the expression of vimentin,α-smooth muscle actin (α-SMA) of HKC cells were assessed by indirect enzyme immunohistochemistry(IEI), and the percentage of α-SMA(+) HKC cells was assessed by flow cytometry.Results No difference in expressions of vimentin and α-SMA by IEI and the percentage of α-SMA(+) HKC cells by cytometry were found between serum-free control HKC cells and Val treated ones.The expressions of vimentin and α-SMA in positive controls were markedly stronger than negative controls;while these expressions in Val+MCP-1+AAI-treated HKC cells were less strong than those in positive controls.The percentage of α-SMA(+) HKC cells in the positive controls was significantly higher than that in negative controls(91.8% vs 3. 1% , P <0. 05),while the percentages of α-SMA(+) HKC cells in MCP-1+AAI + Val group at Val doses of 0. 1,1.0,10.0 pg/ml (17.3%,32.4%,10.0% )were dramatically lower than that in positive controls(P< 0.005).Conclusions (1) No influence of valsartan on transdifferentiation of normal HKC cells in vitro was found in this study.(2) Valsartan may inhibit the transdifferentiation of HKC cells induced by synergistic effect of MCP-I and AAI in vitro.
, 百拇医药
Keywords：Renal tubular epithelial cells;Transdifferentiation; Myofibroblast; Valsartan
基金项目：卫生部研究基金资助项目(98-1-027)
参考文献：
[1]Kuncio GS,Neilson EG Haverty T.Mechanisms tubulointerstitial fibrosis.Kidney Int,1991,39:550-556.
[2]EL Nahas AM.Pathways to renal fibrosis.Exp Nephrol,1995,3:71-75.
[3]Weber KT.Fibrosis,a common pathway to organ failure:angiotensin Ⅱ and tissue repair.Semi Nephrol,1997,17:467 -491.
, 百拇医药
[4]Holstein AF,Maekwaw M,Nagano T,et al.Myofibroblasts in lamina propria of human semi-niferous tubules are dynamic structures of heterogeneous phenotype.Arch Histol Cytol,1996,59:109-125.
[5]Tang WW,Van CY,Qi M.Myofibroblast and α1 collagen expression in experimental tubulointemtitial nephritis.Kidney Int,1997,51:926-931.
[6]Ruizortega M,Egido J.Angiotensin Ⅱ modulates cell growth related events and synthesis of matrix proteins in renal interstitial fibroblasts.Kidney Int,1997,52:1479-1510.
, 百拇医药
[7]Wu LL,Yang N,Roe C J,et al.Macrophage and myofibroblast proliferation in remnant kidney,role of angiotensin Ⅱ.Kidney Int,1997,52 Supple 63:S221-S225.
[8]Klahr S,Morrissey JJ.Comparative study of ACE inhibitor and angiotensin Ⅱ receptor antagonists in interstitial scaring.Kidney Int,1997,52 (S63):S111-S114.
[9]Vanherweghem JL,Depierreux M,Tielemans C,et al.Rapidly progressive interstitial renal fibrosis in young women:association with slimming regimen including Chinese herbs.Lancet,1993,341:387-391.
, http://www.100md.com
[10]Strutz F,Okada H,Cecilia WL,et al.Identification and characterization of a fibroblast marker:FSP1.J Cell Biol,1995,130:393-405.
[11]Okada H,Danoff TM,Kalluri R,et al.Early role of FSP1 in epithelial-mesenchymal transformation.Am J Physiol,1997,273:F563-F574.
[12]Fan JM,Atkins RC,Lan HY.TGFβ1 regulates tubular epithelial myofibroblast transdifferentiation(TEMT) in vitro.Nephrology,1998,4 Suppl:A6.
, http://www.100md.com
[13]Ng YY,Huang TP,Yang WC,et al.Tubular-epithelial-myofibroblast transdifferentiation in progressive tubulointerstitial fibrosis in 5/6 nephrectomized rats.Kidney Int,1998,54:864-876.
[14]Cosyns J,Jadoul M,Squifflet J.Chinese herbs nephropathy:a clue to Balkan epidemic nephropathy? Kidney Int,1994,45:1680-1688.
[15]薄玉红,毕增祺,何祖根,等.中草药马兜铃酸引起的大鼠非坏死肾小管损伤和细胞凋亡初步研究.中华痛脏病杂志,1997,13:168.
收稿日期：1999-10-20, 百拇医药
单位：100730北京，中国医学科学院 中国协和医科大学 北京协和医院肾内科
关键词：肾小管上皮细胞；转分化；肌成纤维细胞；缬沙坦
中华肾病杂志000410摘 要：目的 探讨血管紧张素ⅡⅠ型受体拮抗剂缬沙坦(valsartan，Val)在人类肾小管上皮细胞系(HKC)转分化中的作用。方法 将培养的HKC细胞分为(1)无血清培养对照组；(2)阳性对照组(MCP-1+AAI组)：培养液中加人马兜铃酸-I(AAI)和单核细胞趋化蛋白-1(MCP-1)；(3)Val组：培养液中加入Val；(4)MCP-1+AAI+Val组。应用间接酶标免疫组织化学方法(IEI)检测HKC细胞。Α-平滑肌肌动蛋白(α-SMA)、波形蛋白、角蛋白表达的变化，流式细胞技术检测α-SMA阳性的HKC细胞百分数。结果 不同浓度缬沙坦(0.1、1.0、10.0、100.0pg/ml)分别作用于HKC细胞48h后，IEI测定细胞角蛋白、波形蛋白及α-SMA抗原表达，与无血清对照组比无明显变化；流式细胞术测得Val组表达α-SMA阳性的HKC细胞百分数分别为10.2%、5.6%、5.2%、0.9%，与无血清对照组(平均为3.1%)比较，差异无显著性意义(P>0.05)。MCP-1+AAI+Val(0.1、1.0、10.0pg/ml)组HKC细胞培养48h后，表达α-SMA阳性的HKC细胞分别为17.3%、32.4%、10.0%，与阳性对照组(平均为91.8%)比较，下降非常显著(P均<0.005)。结论 (1)缬沙坦单独作用对正常HKC细胞的转分化无明显影响。(2)缬沙坦对某些因素诱导的HKC细胞转分化可能具有显著抑制作用。
, http://www.100md.com
Inhibition of transdifferentiation of human renal tubular epithelial cells by vaisartan
WEN Xiaoyan ,ZHENG Falei ,SUN Yang
(Division of Nephrology, Peking Union Medical College Hospital,Beijing 100730, China)
Abstract：Objective To investigate the possible role of valsartan(Val) in transdifferentiation of human renal tubular epithelial cell line(HKC).Methods HKC cells were divided into four groups:(1)serum-free(negative control);(2) MCP-1+AAI-treated (positive control);(3) Val-treated alone;(4)Val-inhibition(treated with MCP-1+ AAI+ Val).Then the expression of vimentin,α-smooth muscle actin (α-SMA) of HKC cells were assessed by indirect enzyme immunohistochemistry(IEI), and the percentage of α-SMA(+) HKC cells was assessed by flow cytometry.Results No difference in expressions of vimentin and α-SMA by IEI and the percentage of α-SMA(+) HKC cells by cytometry were found between serum-free control HKC cells and Val treated ones.The expressions of vimentin and α-SMA in positive controls were markedly stronger than negative controls;while these expressions in Val+MCP-1+AAI-treated HKC cells were less strong than those in positive controls.The percentage of α-SMA(+) HKC cells in the positive controls was significantly higher than that in negative controls(91.8% vs 3. 1% , P <0. 05),while the percentages of α-SMA(+) HKC cells in MCP-1+AAI + Val group at Val doses of 0. 1,1.0,10.0 pg/ml (17.3%,32.4%,10.0% )were dramatically lower than that in positive controls(P< 0.005).Conclusions (1) No influence of valsartan on transdifferentiation of normal HKC cells in vitro was found in this study.(2) Valsartan may inhibit the transdifferentiation of HKC cells induced by synergistic effect of MCP-I and AAI in vitro.
, 百拇医药
Keywords：Renal tubular epithelial cells;Transdifferentiation; Myofibroblast; Valsartan
基金项目：卫生部研究基金资助项目(98-1-027)
参考文献：
[1]Kuncio GS,Neilson EG Haverty T.Mechanisms tubulointerstitial fibrosis.Kidney Int,1991,39:550-556.
[2]EL Nahas AM.Pathways to renal fibrosis.Exp Nephrol,1995,3:71-75.
[3]Weber KT.Fibrosis,a common pathway to organ failure:angiotensin Ⅱ and tissue repair.Semi Nephrol,1997,17:467 -491.
, 百拇医药
[4]Holstein AF,Maekwaw M,Nagano T,et al.Myofibroblasts in lamina propria of human semi-niferous tubules are dynamic structures of heterogeneous phenotype.Arch Histol Cytol,1996,59:109-125.
[5]Tang WW,Van CY,Qi M.Myofibroblast and α1 collagen expression in experimental tubulointemtitial nephritis.Kidney Int,1997,51:926-931.
[6]Ruizortega M,Egido J.Angiotensin Ⅱ modulates cell growth related events and synthesis of matrix proteins in renal interstitial fibroblasts.Kidney Int,1997,52:1479-1510.
, 百拇医药
[7]Wu LL,Yang N,Roe C J,et al.Macrophage and myofibroblast proliferation in remnant kidney,role of angiotensin Ⅱ.Kidney Int,1997,52 Supple 63:S221-S225.
[8]Klahr S,Morrissey JJ.Comparative study of ACE inhibitor and angiotensin Ⅱ receptor antagonists in interstitial scaring.Kidney Int,1997,52 (S63):S111-S114.
[9]Vanherweghem JL,Depierreux M,Tielemans C,et al.Rapidly progressive interstitial renal fibrosis in young women:association with slimming regimen including Chinese herbs.Lancet,1993,341:387-391.
, http://www.100md.com
[10]Strutz F,Okada H,Cecilia WL,et al.Identification and characterization of a fibroblast marker:FSP1.J Cell Biol,1995,130:393-405.
[11]Okada H,Danoff TM,Kalluri R,et al.Early role of FSP1 in epithelial-mesenchymal transformation.Am J Physiol,1997,273:F563-F574.
[12]Fan JM,Atkins RC,Lan HY.TGFβ1 regulates tubular epithelial myofibroblast transdifferentiation(TEMT) in vitro.Nephrology,1998,4 Suppl:A6.
, http://www.100md.com
[13]Ng YY,Huang TP,Yang WC,et al.Tubular-epithelial-myofibroblast transdifferentiation in progressive tubulointerstitial fibrosis in 5/6 nephrectomized rats.Kidney Int,1998,54:864-876.
[14]Cosyns J,Jadoul M,Squifflet J.Chinese herbs nephropathy:a clue to Balkan epidemic nephropathy? Kidney Int,1994,45:1680-1688.
[15]薄玉红,毕增祺,何祖根,等.中草药马兜铃酸引起的大鼠非坏死肾小管损伤和细胞凋亡初步研究.中华痛脏病杂志,1997,13:168.
收稿日期：1999-10-20, 百拇医药