中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2009年
6期
420-424
,共5页
叶玲%陈伟英%王欣%李晓艳%唐雪晴%余学清
葉玲%陳偉英%王訢%李曉豔%唐雪晴%餘學清
협령%진위영%왕흔%리효염%당설청%여학청
转化生长囚子β%p38丝裂原活化蛋白激酶类%单核细胞化学吸引蛋白质1%Smad7
轉化生長囚子β%p38絲裂原活化蛋白激酶類%單覈細胞化學吸引蛋白質1%Smad7
전화생장수자β%p38사렬원활화단백격매류%단핵세포화학흡인단백질1%Smad7
Transforming growth factor beta%p38 mitogen-activated protein kinases%Monocyte chemoattractant protein !%Smad7
目的 探讨转化生长因子β1(TGF-β1)在大鼠腹膜间皮细胞(RPMC)中的促炎作用及机制.方法 用TGF-β1(10 μg/L)刺激体外培养的RPMC,观察其对RPMC中单核细胞趋化蛋白1(MCP-1)表达的影响.体外转染Smad7和pcDNA3空载体至RPMC后,观察TGF-β1(10 μg/L)对RPMC MCP-1和p38表达的影响.用p38抑制剂SB203580(10 μmol/L)预处理RPMC后,加入TGF-β1(10 μg/L),观察阻断p38信号通路对MCP-1表达的影响.结果 RT-PCR结果显示,3 h、6 h、12 h、24 h TGF-β1刺激组腹膜间皮细胞MCP-1表达均显著高于0 h对照组(P<0.05),6 h刺激组MCP-1表达达高峰(P<0.01).ELISA结果显示,6 h、12 h、24 h、48 h TGF-β1刺激组MCP-1表达增多(P<0.05),48 h表达达高峰(P<0.01).上调表达Smad7和p38抑制剂SB203580都能明显抑制TGF-β1刺激RPMC产生和分泌MCP-1的作用,与pcDNA3空载体组比较,Smad7治疗组MCP-1的表达显著下调(P<0.05);与TGF-β1刺激组比较,SB203580治疗组MCP-1的表达显著下调(P<0.01).TGF-β1能活化p38磷酸化的过程,上调Smad7可抑制TGF-β1对它们的激活反应;与正常对照组比较,TGF-β1刺激组磷酸化(P)-p38的表达显著上调(P<0.05);与TGF-β1刺激组比较,Smad7治疗组p-p38的表达显著下调(P<0.05).结论 TGF-β1能促进RPMC MCP-1的表达,其促炎作用可能是通过激活p38MAPK信号通路介导的.
目的 探討轉化生長因子β1(TGF-β1)在大鼠腹膜間皮細胞(RPMC)中的促炎作用及機製.方法 用TGF-β1(10 μg/L)刺激體外培養的RPMC,觀察其對RPMC中單覈細胞趨化蛋白1(MCP-1)錶達的影響.體外轉染Smad7和pcDNA3空載體至RPMC後,觀察TGF-β1(10 μg/L)對RPMC MCP-1和p38錶達的影響.用p38抑製劑SB203580(10 μmol/L)預處理RPMC後,加入TGF-β1(10 μg/L),觀察阻斷p38信號通路對MCP-1錶達的影響.結果 RT-PCR結果顯示,3 h、6 h、12 h、24 h TGF-β1刺激組腹膜間皮細胞MCP-1錶達均顯著高于0 h對照組(P<0.05),6 h刺激組MCP-1錶達達高峰(P<0.01).ELISA結果顯示,6 h、12 h、24 h、48 h TGF-β1刺激組MCP-1錶達增多(P<0.05),48 h錶達達高峰(P<0.01).上調錶達Smad7和p38抑製劑SB203580都能明顯抑製TGF-β1刺激RPMC產生和分泌MCP-1的作用,與pcDNA3空載體組比較,Smad7治療組MCP-1的錶達顯著下調(P<0.05);與TGF-β1刺激組比較,SB203580治療組MCP-1的錶達顯著下調(P<0.01).TGF-β1能活化p38燐痠化的過程,上調Smad7可抑製TGF-β1對它們的激活反應;與正常對照組比較,TGF-β1刺激組燐痠化(P)-p38的錶達顯著上調(P<0.05);與TGF-β1刺激組比較,Smad7治療組p-p38的錶達顯著下調(P<0.05).結論 TGF-β1能促進RPMC MCP-1的錶達,其促炎作用可能是通過激活p38MAPK信號通路介導的.
목적 탐토전화생장인자β1(TGF-β1)재대서복막간피세포(RPMC)중적촉염작용급궤제.방법 용TGF-β1(10 μg/L)자격체외배양적RPMC,관찰기대RPMC중단핵세포추화단백1(MCP-1)표체적영향.체외전염Smad7화pcDNA3공재체지RPMC후,관찰TGF-β1(10 μg/L)대RPMC MCP-1화p38표체적영향.용p38억제제SB203580(10 μmol/L)예처리RPMC후,가입TGF-β1(10 μg/L),관찰조단p38신호통로대MCP-1표체적영향.결과 RT-PCR결과현시,3 h、6 h、12 h、24 h TGF-β1자격조복막간피세포MCP-1표체균현저고우0 h대조조(P<0.05),6 h자격조MCP-1표체체고봉(P<0.01).ELISA결과현시,6 h、12 h、24 h、48 h TGF-β1자격조MCP-1표체증다(P<0.05),48 h표체체고봉(P<0.01).상조표체Smad7화p38억제제SB203580도능명현억제TGF-β1자격RPMC산생화분비MCP-1적작용,여pcDNA3공재체조비교,Smad7치료조MCP-1적표체현저하조(P<0.05);여TGF-β1자격조비교,SB203580치료조MCP-1적표체현저하조(P<0.01).TGF-β1능활화p38린산화적과정,상조Smad7가억제TGF-β1대타문적격활반응;여정상대조조비교,TGF-β1자격조린산화(P)-p38적표체현저상조(P<0.05);여TGF-β1자격조비교,Smad7치료조p-p38적표체현저하조(P<0.05).결론 TGF-β1능촉진RPMC MCP-1적표체,기촉염작용가능시통과격활p38MAPK신호통로개도적.
Objective To investigate the pro-inflammatory effect of transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells (RPMCs) and its machanism. Methods TGF-β1-induced RPMCs model in vitro was established, and the expression of MCP-1 in the TGF-β1-induced RPMCs was observed. The intervention of Smad7 on the expression of MCP-1 and p38 signal proteins induced by TGF-β1 in RPMCs was explored as well as the intervention of p38 inhibitor SB203580 on the expression of MCP-1 induced by TGF-β1 in RPMCs. Results TGF-β1 could stimulate MCP-1 expression in RPMCs. Compared with control group, MCP-1 mRNA levels were significantly increased after 3 h treatment with TGF-β1 (P<0.05), peak MCP-1 induction occurred at 6 h (P<0.01), and the stimulatory effect of TGF-β1 persisted through 24 h (P<0.05). MCP-1 protein levels were significantly increased after 6 h treatment with TGF-β1(P<0.05), peak MCP-1 induction occurred at 48 h(P<0.01). Over-expressed Smad7 and p38 inhibitor could reduce the expression of MCP-1 induced by TGF-β1 (P<0.05). TGF-β1 could activate p38 signaling pathway, but over-expressed Smad7 could inhibit this role of TGF-β1. Compared with control group, the expression level of p-p38 was increased in TGF-β1-stimulated group. Compared with TGF-β1-stimulated group, the expression level of p-p38 was reduced in Smad7 gene transfer group. Conclusions TGF-β1-induced MCP-1 expression in rat peritoneal mesothelial cells is p38MAPK dependent.
