中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2013年
34期
2750-2754
,共5页
哮喘%细胞质膜微囊蛋白%肌细胞,平滑肌%细胞增殖%罗红霉素
哮喘%細胞質膜微囊蛋白%肌細胞,平滑肌%細胞增殖%囉紅黴素
효천%세포질막미낭단백%기세포,평활기%세포증식%라홍매소
Asthma%Caveolins%Myocytes,smooth muscle%Cell proliferation%Roxithromycin
目的 探讨微囊蛋白1抑制气道平滑肌细胞(ASMCs)增殖的可能机制以及罗红霉素的调控作用.方法 复制哮喘大鼠模型,光镜观察肺组织病理学变化,透射电镜观察各组ASMCs上微囊的结构及变化,用组织贴壁法体外培养ASMCs,实验设对照组(A组:含2.5％胎牛血清的高糖培养液处理1h)、哮喘组(B组:处理同A组)、细胞外调节蛋白激酶(ERK1/2)信号通路阻断剂PD98059组(C组:10×10-6mol/L的PD98059处理1h)、罗红霉素组(D组:100 mg/L的罗红霉素处理1h)、β-甲基环糊精组(E组:5μmol/L的β甲基环糊精处理1h)；用细胞计数试剂盒(CCK-8)检测各组细胞的增殖情况,Western印迹检测各组微囊蛋白1表达；逆转录(RT)-PCR和Western印迹分别检测ERK1/2mRNA、单核细胞趋化蛋白1(MCP-1)mRNA和磷酸化ERK1/2、MCP-1蛋白的表达.结果 C、D组ASMCs增殖显著低于B组(0.68±0.15、0.63 ±0.13比0.96 ±0.14,均P＜0.05)；E组(1.26±0.11) ASMCs增殖强于B组(P＜0.05).C、D组微囊蛋白1表达均显著高于B组(0.332±0.057、0.392±0.064比0.237±0.032,均P＜0.05)；C、D组ERK1/2蛋白表达均显著低于B组(0.241±0.017、0.268±0.007比0.346±0.009,均P＜0.01),E组(0.441±0.011) ERK1/2蛋白表达高于B组；C、D组ERK1/2 mRNA表达均显著低于B组(0.277±0.043、0.338±0.026比0.591±0.022,均P＜0.01).C、D组MCP-1蛋白表达均显著低于B组(0.198±0.015、0.286±0.019比0.482±0.026,均P＜0.01),E组(0.521±0.023) MCP-1蛋白表达较B有升高趋势；C、D组MCP-1mRNA表达均显著低于B组(0.212 ±0.042、0.249±0.032比0.676±0.053,均P＜0.01).结论 微囊蛋白1可能通过ERK信号通路抑制MCP-1表达,从而抑制哮喘ASMCs的增殖.罗红霉素可能上调微囊蛋白1的表达,抑制MCP-1 mRNA表达和翻译,进而抑制哮喘ASMCs的增殖.
目的 探討微囊蛋白1抑製氣道平滑肌細胞(ASMCs)增殖的可能機製以及囉紅黴素的調控作用.方法 複製哮喘大鼠模型,光鏡觀察肺組織病理學變化,透射電鏡觀察各組ASMCs上微囊的結構及變化,用組織貼壁法體外培養ASMCs,實驗設對照組(A組:含2.5％胎牛血清的高糖培養液處理1h)、哮喘組(B組:處理同A組)、細胞外調節蛋白激酶(ERK1/2)信號通路阻斷劑PD98059組(C組:10×10-6mol/L的PD98059處理1h)、囉紅黴素組(D組:100 mg/L的囉紅黴素處理1h)、β-甲基環糊精組(E組:5μmol/L的β甲基環糊精處理1h)；用細胞計數試劑盒(CCK-8)檢測各組細胞的增殖情況,Western印跡檢測各組微囊蛋白1錶達；逆轉錄(RT)-PCR和Western印跡分彆檢測ERK1/2mRNA、單覈細胞趨化蛋白1(MCP-1)mRNA和燐痠化ERK1/2、MCP-1蛋白的錶達.結果 C、D組ASMCs增殖顯著低于B組(0.68±0.15、0.63 ±0.13比0.96 ±0.14,均P＜0.05)；E組(1.26±0.11) ASMCs增殖彊于B組(P＜0.05).C、D組微囊蛋白1錶達均顯著高于B組(0.332±0.057、0.392±0.064比0.237±0.032,均P＜0.05)；C、D組ERK1/2蛋白錶達均顯著低于B組(0.241±0.017、0.268±0.007比0.346±0.009,均P＜0.01),E組(0.441±0.011) ERK1/2蛋白錶達高于B組；C、D組ERK1/2 mRNA錶達均顯著低于B組(0.277±0.043、0.338±0.026比0.591±0.022,均P＜0.01).C、D組MCP-1蛋白錶達均顯著低于B組(0.198±0.015、0.286±0.019比0.482±0.026,均P＜0.01),E組(0.521±0.023) MCP-1蛋白錶達較B有升高趨勢；C、D組MCP-1mRNA錶達均顯著低于B組(0.212 ±0.042、0.249±0.032比0.676±0.053,均P＜0.01).結論 微囊蛋白1可能通過ERK信號通路抑製MCP-1錶達,從而抑製哮喘ASMCs的增殖.囉紅黴素可能上調微囊蛋白1的錶達,抑製MCP-1 mRNA錶達和翻譯,進而抑製哮喘ASMCs的增殖.
목적 탐토미낭단백1억제기도평활기세포(ASMCs)증식적가능궤제이급라홍매소적조공작용.방법 복제효천대서모형,광경관찰폐조직병이학변화,투사전경관찰각조ASMCs상미낭적결구급변화,용조직첩벽법체외배양ASMCs,실험설대조조(A조:함2.5％태우혈청적고당배양액처리1h)、효천조(B조:처리동A조)、세포외조절단백격매(ERK1/2)신호통로조단제PD98059조(C조:10×10-6mol/L적PD98059처리1h)、라홍매소조(D조:100 mg/L적라홍매소처리1h)、β-갑기배호정조(E조:5μmol/L적β갑기배호정처리1h)；용세포계수시제합(CCK-8)검측각조세포적증식정황,Western인적검측각조미낭단백1표체；역전록(RT)-PCR화Western인적분별검측ERK1/2mRNA、단핵세포추화단백1(MCP-1)mRNA화린산화ERK1/2、MCP-1단백적표체.결과 C、D조ASMCs증식현저저우B조(0.68±0.15、0.63 ±0.13비0.96 ±0.14,균P＜0.05)；E조(1.26±0.11) ASMCs증식강우B조(P＜0.05).C、D조미낭단백1표체균현저고우B조(0.332±0.057、0.392±0.064비0.237±0.032,균P＜0.05)；C、D조ERK1/2단백표체균현저저우B조(0.241±0.017、0.268±0.007비0.346±0.009,균P＜0.01),E조(0.441±0.011) ERK1/2단백표체고우B조；C、D조ERK1/2 mRNA표체균현저저우B조(0.277±0.043、0.338±0.026비0.591±0.022,균P＜0.01).C、D조MCP-1단백표체균현저저우B조(0.198±0.015、0.286±0.019비0.482±0.026,균P＜0.01),E조(0.521±0.023) MCP-1단백표체교B유승고추세；C、D조MCP-1mRNA표체균현저저우B조(0.212 ±0.042、0.249±0.032비0.676±0.053,균P＜0.01).결론 미낭단백1가능통과ERK신호통로억제MCP-1표체,종이억제효천ASMCs적증식.라홍매소가능상조미낭단백1적표체,억제MCP-1 mRNA표체화번역,진이억제효천ASMCs적증식.
Objective To explore the functional role of caveolin-1 in airway smooth muscle cells (ASMCs) proliferation and examine the regulatory effect of roxithromycin.Methods The rat model of bronchial asthma was established.Electron microscope was employed to observe the status of caveolae and light microscope for the histological changes in pulmonary tissues.The primarily cultured ASMCs were divided into 5 groups:control (group A),asthmatic ASMCs (group B),PD98059 (group C),roxithroymcin (group D) and methyl-β-cyclodextrin (group E).Cell proliferation was detected by Cell Counting Kit-8 (CCK-8).And the expressions of caveolin-1,extracellular regulated protein kinases (ERK) and monocyte chemotactic protein (MCP)-1 were detected by Western blot and reverse transcription polymerase chain reaction (RT-PCR).Results The cell proliferation of asthmatic ASMCs (0.68 ± 0.15,0.63 ±0.13) in groups C and D were significantly less than those in group B (0.96 ±0.14) (both P ＜0.05) while group E was more than group B (1.26 ± 0.11 vs 0.96 ± 0.14,P ＜ 0.05).The content of caveolin-1 (0.392 ±0.064,0.332 ±0.057) in groups C and D were higher than those in group B (0.237 ±0.032) (both P ＜0.05) while ERK1/2 protein level in groups C and D (0.241 ±0.017,0.268 ±0.007) were less than those in group B (0.346 ±0.009) (both P ＜0.01).And MCP-1 protein level in groups C and D (0.198 ±0.015,0.286 ±0.019) were less than those in group B (0.482 ±0.026) (both P ＜0.01).The ERK mRNA level in groups C and D (0.277 ±0.043,0.338 ±0.026) were less than those in group B (0.591 ±0.022) (both P＜0.01).And also MCP-1 mRNA in groups C and D (0.212 ±0.042,0.249 ± 0.032) were less than those in group B (0.676 ± 0.053) (all P ＜ 0.01).Conclusions Caveolin-1 preventing the proliferation of asthmatic ASMCs is most likely mediated by ERK1/2 signal pathway and a down-regulation of MCP-1 expression.And roxithroymcin reduces the proliferation of asthmatic ASMCs through up-regulating the expression of caveolin-1 and inhibiting the expression of MCP-1.
