中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2008年
11期
832-837
,共6页
抵抗素%肾小球系膜细胞%p38丝裂原活化蛋白激酶类%增殖
牴抗素%腎小毬繫膜細胞%p38絲裂原活化蛋白激酶類%增殖
저항소%신소구계막세포%p38사렬원활화단백격매류%증식
Resistin%Mesangial cells%p38 mitogen-activated protein kinases%Proliferation
目的 观察巨噬细胞因子抵抗素过度表达对高糖刺激作用下人肾小球系膜细胞p38丝裂原活化蛋白激酶(MAPK)信号通路的影响,探讨抵抗素调控肾小球系膜细胞增殖及细胞外基质积聚的作用机制.方法 通过转染携带野牛型抵抗素基因的腺病毒载体(Ad-resistin)构建过度表达抵抗素的人巨噬细胞模型,并与高糖刺激后的人肾小球系膜细胞共培养.3H-氚标胸腺嘧啶掺入法检测.肾小球系膜细胞增殖.免疫细胞化学法检测系膜细胞激活蛋白1(AP-1)的表达.免疫荧光检测细胞外基质层粘连蛋白(LN)的表达.Western印迹检测系膜细胞内p38MAPK、转化牛长因子(TGF)IM的表达并测定Smad2的磷酸化水平.结果 Ad-resistin转染后,人巨噬细胞抵抗素mRNA水平及蛋白表达均显著升高(P<0-01).与对照组比较,与过度表达抵抗素的人巨噬细胞共培养后,人肾小球系膜细胞p38MAPK、TGF-β1的蛋白表达显著增强;细胞内Smad2的磷酸化水平显著升高(P<0.05);肾小球系膜细胞出现明显的增殖(P<0.01);细胞外基质的合成增多(P<0.05).结论 巨噬细胞因子抵抗素的过度表达可能通过p38MAPK信号通路,促进高糖刺激作用下肾小球系膜细胞的增殖及细胞外基质的异常积聚.
目的 觀察巨噬細胞因子牴抗素過度錶達對高糖刺激作用下人腎小毬繫膜細胞p38絲裂原活化蛋白激酶(MAPK)信號通路的影響,探討牴抗素調控腎小毬繫膜細胞增殖及細胞外基質積聚的作用機製.方法 通過轉染攜帶野牛型牴抗素基因的腺病毒載體(Ad-resistin)構建過度錶達牴抗素的人巨噬細胞模型,併與高糖刺激後的人腎小毬繫膜細胞共培養.3H-氚標胸腺嘧啶摻入法檢測.腎小毬繫膜細胞增殖.免疫細胞化學法檢測繫膜細胞激活蛋白1(AP-1)的錶達.免疫熒光檢測細胞外基質層粘連蛋白(LN)的錶達.Western印跡檢測繫膜細胞內p38MAPK、轉化牛長因子(TGF)IM的錶達併測定Smad2的燐痠化水平.結果 Ad-resistin轉染後,人巨噬細胞牴抗素mRNA水平及蛋白錶達均顯著升高(P<0-01).與對照組比較,與過度錶達牴抗素的人巨噬細胞共培養後,人腎小毬繫膜細胞p38MAPK、TGF-β1的蛋白錶達顯著增彊;細胞內Smad2的燐痠化水平顯著升高(P<0.05);腎小毬繫膜細胞齣現明顯的增殖(P<0.01);細胞外基質的閤成增多(P<0.05).結論 巨噬細胞因子牴抗素的過度錶達可能通過p38MAPK信號通路,促進高糖刺激作用下腎小毬繫膜細胞的增殖及細胞外基質的異常積聚.
목적 관찰거서세포인자저항소과도표체대고당자격작용하인신소구계막세포p38사렬원활화단백격매(MAPK)신호통로적영향,탐토저항소조공신소구계막세포증식급세포외기질적취적작용궤제.방법 통과전염휴대야우형저항소기인적선병독재체(Ad-resistin)구건과도표체저항소적인거서세포모형,병여고당자격후적인신소구계막세포공배양.3H-천표흉선밀정참입법검측.신소구계막세포증식.면역세포화학법검측계막세포격활단백1(AP-1)적표체.면역형광검측세포외기질층점련단백(LN)적표체.Western인적검측계막세포내p38MAPK、전화우장인자(TGF)IM적표체병측정Smad2적린산화수평.결과 Ad-resistin전염후,인거서세포저항소mRNA수평급단백표체균현저승고(P<0-01).여대조조비교,여과도표체저항소적인거서세포공배양후,인신소구계막세포p38MAPK、TGF-β1적단백표체현저증강;세포내Smad2적린산화수평현저승고(P<0.05);신소구계막세포출현명현적증식(P<0.01);세포외기질적합성증다(P<0.05).결론 거서세포인자저항소적과도표체가능통과p38MAPK신호통로,촉진고당자격작용하신소구계막세포적증식급세포외기질적이상적취.
Objective To investigate the effects of resistin on mesangial cells proliferation induced by high glucose and subsequent change of p38MAPK signal pathway. Methods Human macrophrages were cultured and treated with adenovirus encoding for resistin (Ad-resistin) for 48 h and were then co-cultured with human mesangial cells stimulated by high glucose for another 48 h. Mesangial ceils were harvested and their proliferation was measured by 3H-TdR. Activator protein 1 (AP-1) was examined by immunocytochemistry and laminin of excellular matrix was observed with immuofluorescence. Protein levels of p38MAPK and TGF-β1 were measured by Western blot. Smad2 phosphatase activity was aslo detected by Western blot. Results The mRNA and protein levels of resistin were significantly higher in Ad-resistin treated macrophages than those in Ad treated cells (P<0.01). Over-expression of resistin up-regulated p38MAPK protein levels of human mesangial cells(P<0.05). Resistin also promoted the proliferation of mesangial cells (P<0.01) and the synthesis of laminin stimulated by high glucose. The expression of TGF-β1 and phosphorylation of Smad2 were up-regulated in the mesangial cells (P< 0.05). Conclusion Macrophage cytokine resistin may promote mesangial cells proliferation and abnormal accumulation of excellular matrix stimulated by high glucose via activating p38MAPK signal passway.
