中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2011年
12期
928-932
,共5页
JNK丝裂原活化蛋白激酶%连接蛋白43%细胞系,转化%肾小管上皮细胞
JNK絲裂原活化蛋白激酶%連接蛋白43%細胞繫,轉化%腎小管上皮細胞
JNK사렬원활화단백격매%련접단백43%세포계,전화%신소관상피세포
JNK mitogen-activated protein kinase%Connexin 43%Cell line transformed%Renal tubular epithelial cells
目的 观察c-Jun氨基末端激酶(JNK)-c-Jun通路对连接蛋白43(Cx43)表达的影响及在转化生长因子(TGF)β1诱导的肾小管上皮细胞-肌成纤维细胞转分化(TEMT)中的作用.方法 大鼠肾小管上皮细胞(NRK-52E)随机分成3组:对照组、TGF-β1(10μg/L)组和TGF-β1(10 μg/L)+JNK选择性抑制剂SP600125(50 μmol/L)组.用免疫细胞化学、Western印迹检测JNK、c -Jun、连接蛋白43( Cx43)、上皮细胞标志物E-钙黏蛋白(E-cadherin)和肌成纤维细胞标志物α-SMA的表达.用RT-PCR检测Cx43的mRNA水平.用激光共聚焦显微镜荧光漂白恢复( FRAP)技术检测NRK-52E细胞间通讯功能.结果 TGF-β1引起肾小管上皮细胞α-SMA、JNK、c-Jun表达上调(均P＜0.05),Cx43、E-cadherin表达下调(均P＜0.05),Cx43的mRNA水平下降(P＜0.05),细胞间通迅功能下降(P＜0.05).JNK抑制剂处理后,上述改变明显减轻.结论 TGF-β1引起肾小管上皮细胞内JNK表达上调,增加c-Jun活性,从而抑制Cx43的表达和降低细胞间通迅功能,导致TEMT.
目的 觀察c-Jun氨基末耑激酶(JNK)-c-Jun通路對連接蛋白43(Cx43)錶達的影響及在轉化生長因子(TGF)β1誘導的腎小管上皮細胞-肌成纖維細胞轉分化(TEMT)中的作用.方法 大鼠腎小管上皮細胞(NRK-52E)隨機分成3組:對照組、TGF-β1(10μg/L)組和TGF-β1(10 μg/L)+JNK選擇性抑製劑SP600125(50 μmol/L)組.用免疫細胞化學、Western印跡檢測JNK、c -Jun、連接蛋白43( Cx43)、上皮細胞標誌物E-鈣黏蛋白(E-cadherin)和肌成纖維細胞標誌物α-SMA的錶達.用RT-PCR檢測Cx43的mRNA水平.用激光共聚焦顯微鏡熒光漂白恢複( FRAP)技術檢測NRK-52E細胞間通訊功能.結果 TGF-β1引起腎小管上皮細胞α-SMA、JNK、c-Jun錶達上調(均P＜0.05),Cx43、E-cadherin錶達下調(均P＜0.05),Cx43的mRNA水平下降(P＜0.05),細胞間通迅功能下降(P＜0.05).JNK抑製劑處理後,上述改變明顯減輕.結論 TGF-β1引起腎小管上皮細胞內JNK錶達上調,增加c-Jun活性,從而抑製Cx43的錶達和降低細胞間通迅功能,導緻TEMT.
목적 관찰c-Jun안기말단격매(JNK)-c-Jun통로대련접단백43(Cx43)표체적영향급재전화생장인자(TGF)β1유도적신소관상피세포-기성섬유세포전분화(TEMT)중적작용.방법 대서신소관상피세포(NRK-52E)수궤분성3조:대조조、TGF-β1(10μg/L)조화TGF-β1(10 μg/L)+JNK선택성억제제SP600125(50 μmol/L)조.용면역세포화학、Western인적검측JNK、c -Jun、련접단백43( Cx43)、상피세포표지물E-개점단백(E-cadherin)화기성섬유세포표지물α-SMA적표체.용RT-PCR검측Cx43적mRNA수평.용격광공취초현미경형광표백회복( FRAP)기술검측NRK-52E세포간통신공능.결과 TGF-β1인기신소관상피세포α-SMA、JNK、c-Jun표체상조(균P＜0.05),Cx43、E-cadherin표체하조(균P＜0.05),Cx43적mRNA수평하강(P＜0.05),세포간통신공능하강(P＜0.05).JNK억제제처리후,상술개변명현감경.결론 TGF-β1인기신소관상피세포내JNK표체상조,증가c-Jun활성,종이억제Cx43적표체화강저세포간통신공능,도치TEMT.
Objective To explore the effect of JNK-c-Jun signal pathway on connexin 43 (Cx43) expression and its role in renal tubular epithelial-myofibroblast transition (TEMT) induced by TGF-β1. Methods Normal rat kidney tubular epithelial cells (NRK-52E) were cultured in Dulbecco's modified eagle medium (DMEM) with 10％ fetal bovine serum,then were randomly divided into 3 groups: control group,TGF-β1 group (treated with TGF-β1 10 μg/L),and TGF-β1+SP600125 (selective JNK inhibitor,50 μmol/L) group. The protein expressions of JNK,c-Jun,α-SMA,Cx43 and E-cadherin were assayed by immunocytochemistry and Western blotting.The Cx43mRNA was assayed by RT-PCR.Gap junction intercellular communication (CJIC) was measured by fluorescence recovery after photobleaching assay (FRAP). Results TGF-β1 increased the expressions of JNK,c-Jun and α-SMA (P＜0.05),reduced the expressions of Cx43 and E-cadherin (P＜0.05),and inhibited GJIC of NRK-52E (P ＜0.05).SP600125 could alleviate the above expressions changes and enhanced GJIC induced by TGF-β1. Conclusion JNK-c-Jun signal pathway induces TEMT of NRK-52E treated with TGF-β1 via down-regulation of connexin 43expression and inhibition of GJIC.
