中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2014年
8期
598-603
,共6页
仇芝%陈晓岚%徐玉音%潘天昳%马雯%范亚平
仇芝%陳曉嵐%徐玉音%潘天昳%馬雯%範亞平
구지%진효람%서옥음%반천질%마문%범아평
受体,前列腺素E%转化生长因子β1%系膜细胞%拮抗剂%细胞外信号调节蛋白激酶1/2
受體,前列腺素E%轉化生長因子β1%繫膜細胞%拮抗劑%細胞外信號調節蛋白激酶1/2
수체,전렬선소E%전화생장인자β1%계막세포%길항제%세포외신호조절단백격매1/2
Receptors,prostaglandin E%Transforming growth factor-beta 1%Mesangial cells%Antagonist%Extracellular signal-regulated protein kinase 1/2
目的 探讨前列腺素E2(PGE2)受体1亚型(EP1)拮抗剂(SC-19220)对转化生长因子(TGF)β1诱导的小鼠肾小球系膜细胞增殖、前列腺素合酶表达、细胞外调节蛋白激酶(ERK)信号通路的影响及可能机制.方法 小鼠肾小球系膜细胞分组:对照组;TGF-β1(10μg/L)组;药物干预组:不同浓度(0.1、0.5、1.0 μmol/L)SC-19220+ TGF-β1(10卜Lg/L)组.CCK-8法检测细胞增殖;ELISA法检测细胞上清中PGE2的表达;实时荧光定量PCR法检测细胞结缔组织生长因子(CTGF)、层粘连蛋白(LN)、环氧化酶2(COX2)、膜结合型前列腺素E2合酶1(mPGES1) mRNA的表达.Western印迹法检测CTGF、LN、COX2、mPGES1及ERK1/2活性变化.结果 与对照组比,TGF-β1组系膜细胞增殖增加(P ＜ 0.05),PGE2表达增加(P ＜ 0.05),CTGF、LN、COX2、mPGES1 mRNA及蛋白表达增加(P＜0.05),ERK1/2活性增加(P＜0.05).SC-19220干预后呈剂量依赖性抑制系膜细胞增殖(P＜0.05),下调PGE2的表达(P ＜ 0.05),抑制CTGF、LN、COX2、mPGES1 mRNA及蛋白表达(P ＜ 0.05),抑制ERK1/2活性(P＜0.05).结论 SC-19220可能通过抑制ERK1/2活性,反馈抑制COX2、mPGES1及PGE2的表达,从而下调CTGF、LN的表达,减轻TGF-β1诱导的系膜细胞损伤.
目的 探討前列腺素E2(PGE2)受體1亞型(EP1)拮抗劑(SC-19220)對轉化生長因子(TGF)β1誘導的小鼠腎小毬繫膜細胞增殖、前列腺素閤酶錶達、細胞外調節蛋白激酶(ERK)信號通路的影響及可能機製.方法 小鼠腎小毬繫膜細胞分組:對照組;TGF-β1(10μg/L)組;藥物榦預組:不同濃度(0.1、0.5、1.0 μmol/L)SC-19220+ TGF-β1(10蔔Lg/L)組.CCK-8法檢測細胞增殖;ELISA法檢測細胞上清中PGE2的錶達;實時熒光定量PCR法檢測細胞結締組織生長因子(CTGF)、層粘連蛋白(LN)、環氧化酶2(COX2)、膜結閤型前列腺素E2閤酶1(mPGES1) mRNA的錶達.Western印跡法檢測CTGF、LN、COX2、mPGES1及ERK1/2活性變化.結果 與對照組比,TGF-β1組繫膜細胞增殖增加(P ＜ 0.05),PGE2錶達增加(P ＜ 0.05),CTGF、LN、COX2、mPGES1 mRNA及蛋白錶達增加(P＜0.05),ERK1/2活性增加(P＜0.05).SC-19220榦預後呈劑量依賴性抑製繫膜細胞增殖(P＜0.05),下調PGE2的錶達(P ＜ 0.05),抑製CTGF、LN、COX2、mPGES1 mRNA及蛋白錶達(P ＜ 0.05),抑製ERK1/2活性(P＜0.05).結論 SC-19220可能通過抑製ERK1/2活性,反饋抑製COX2、mPGES1及PGE2的錶達,從而下調CTGF、LN的錶達,減輕TGF-β1誘導的繫膜細胞損傷.
목적 탐토전렬선소E2(PGE2)수체1아형(EP1)길항제(SC-19220)대전화생장인자(TGF)β1유도적소서신소구계막세포증식、전렬선소합매표체、세포외조절단백격매(ERK)신호통로적영향급가능궤제.방법 소서신소구계막세포분조:대조조;TGF-β1(10μg/L)조;약물간예조:불동농도(0.1、0.5、1.0 μmol/L)SC-19220+ TGF-β1(10복Lg/L)조.CCK-8법검측세포증식;ELISA법검측세포상청중PGE2적표체;실시형광정량PCR법검측세포결체조직생장인자(CTGF)、층점련단백(LN)、배양화매2(COX2)、막결합형전렬선소E2합매1(mPGES1) mRNA적표체.Western인적법검측CTGF、LN、COX2、mPGES1급ERK1/2활성변화.결과 여대조조비,TGF-β1조계막세포증식증가(P ＜ 0.05),PGE2표체증가(P ＜ 0.05),CTGF、LN、COX2、mPGES1 mRNA급단백표체증가(P＜0.05),ERK1/2활성증가(P＜0.05).SC-19220간예후정제량의뢰성억제계막세포증식(P＜0.05),하조PGE2적표체(P ＜ 0.05),억제CTGF、LN、COX2、mPGES1 mRNA급단백표체(P ＜ 0.05),억제ERK1/2활성(P＜0.05).결론 SC-19220가능통과억제ERK1/2활성,반궤억제COX2、mPGES1급PGE2적표체,종이하조CTGF、LN적표체,감경TGF-β1유도적계막세포손상.
Objective To explore the effects and mechanisms of prostaglandin E2 (PGE2) receptor 1 antagonist (SC-19220) on proliferation,prostaglandin synthase and extracellular regulated protein kinases (ERK) signal pathway induced by transforming growth factor β1(TGF-β1) in glomerular mesangial cells.Methods Mouse glomerular mesangial cells (GMCs) were divided into 5 groups:control group,TGF-β1 (10 μg/L) group,TGF-β1 (10 μg/L) plus SC-19220 group (0.1,0.5,1.0 μmol/L).The proliferation of GMCs was measured by CCK-8.The PGE2 in supernatant was measured by ELISA.The expression of connective tissue growth factor (CTGF),laminin (LN),cyclooxygenase 2(COX2),membrane-bound prostaglandin E2 synthase 1 (mPGES1) protein and mRNA was examined by Westem blotting and real-time quantitative PCR,ERK1/2 or phospho-ERK1/2 was measured by Western blotting as well.Results TGF-β1 induced the proliferation of GMCs and increased the secretion of PGE2.Besides,TGF-β1 significantly up-regulated the expression of CTGF,LN,COX2 and mPGES1 mRNA and protein (P ＜ 0.05),and increased the expression of phospho-ERK1/2 protein (P ＜ 0.05).However,SC-19220 significantly attenuated the changes of above-mentioned parameters and their activities (P ＜ 0.05).All the effects of SC-19220 were in dose-dependent manner.Conclusions SC19220 may reduce TGF-β1-induced cell damage by suppressing the activity of ERK1/2,and feedback inhibition of COX2,mPGES1 and PGE2,thus decreases the expression of LN and CTGF.
