天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2015年
9期
978-980,981
,共4页
管频%于化鹏%吴智勇%李伟%吴洁
管頻%于化鵬%吳智勇%李偉%吳潔
관빈%우화붕%오지용%리위%오길
肌细胞,平滑肌%细胞增殖%肺疾病,慢性阻塞性%体外研究%香烟提取物%钙网织蛋白%CCAAT增强子结合蛋白
肌細胞,平滑肌%細胞增殖%肺疾病,慢性阻塞性%體外研究%香煙提取物%鈣網織蛋白%CCAAT增彊子結閤蛋白
기세포,평활기%세포증식%폐질병,만성조새성%체외연구%향연제취물%개망직단백%CCAAT증강자결합단백
myocytes,smooth muscle%cell proliferation%pulmonary disease,chronic obstructive%in vitro%cigarette smoke extract%calreticulin%CCAAT enhancer-binding protein
目的：探讨香烟提取物（CSE）对人气道平滑肌细胞（ASMCs）增殖以及钙网织蛋白（CRT）、CCAAT增强子结合蛋白α（CEBPα）表达的影响及机制。方法（1）通过收集经不同浓度CSE处理24 h的ASMCs，分为对照组、2.5%CSE组、5%CSE组、10%CSE组。以MTT法分析各组细胞增殖情况，采用逆转录聚合酶链反应（RT-PCR）检测各组细胞CEBPα的mRNA水平；免疫印迹法检测4组细胞CRT、CEBPα的蛋白水平。（2）在10%CSE组，分别在ASMCs中转染阴性对照siRNA、CRT的siRNA，比较2组ASMCs的CEBPα、CRT表达及细胞增殖情况。结果（1）对照组、2.5％CSE组、5％CSE组、10％CSE组ASMCs的CEBPα蛋白表达依次递减，CRT和细胞增殖程度呈依次增高（P＜0.05）；而CEBPαmRNA表达差异无统计学意义（P＞0.05）。（2）在10％CSE作用下，CRTsiRNA组中ASMCs的CEBPα表达较阴性对照siRNA组增强（P＜0.05），而CRT和细胞增殖程度均较相应阴性对照siRNA组减弱（P＜0.05）。结论 CSE可使ASMCs中CRT的表达增强，从而抑制CEBPαmRNA的翻译，使CEBPα的表达减少，最终促进ASMCs的增殖。
目的：探討香煙提取物（CSE）對人氣道平滑肌細胞（ASMCs）增殖以及鈣網織蛋白（CRT）、CCAAT增彊子結閤蛋白α（CEBPα）錶達的影響及機製。方法（1）通過收集經不同濃度CSE處理24 h的ASMCs，分為對照組、2.5%CSE組、5%CSE組、10%CSE組。以MTT法分析各組細胞增殖情況，採用逆轉錄聚閤酶鏈反應（RT-PCR）檢測各組細胞CEBPα的mRNA水平；免疫印跡法檢測4組細胞CRT、CEBPα的蛋白水平。（2）在10%CSE組，分彆在ASMCs中轉染陰性對照siRNA、CRT的siRNA，比較2組ASMCs的CEBPα、CRT錶達及細胞增殖情況。結果（1）對照組、2.5％CSE組、5％CSE組、10％CSE組ASMCs的CEBPα蛋白錶達依次遞減，CRT和細胞增殖程度呈依次增高（P＜0.05）；而CEBPαmRNA錶達差異無統計學意義（P＞0.05）。（2）在10％CSE作用下，CRTsiRNA組中ASMCs的CEBPα錶達較陰性對照siRNA組增彊（P＜0.05），而CRT和細胞增殖程度均較相應陰性對照siRNA組減弱（P＜0.05）。結論 CSE可使ASMCs中CRT的錶達增彊，從而抑製CEBPαmRNA的翻譯，使CEBPα的錶達減少，最終促進ASMCs的增殖。
목적：탐토향연제취물（CSE）대인기도평활기세포（ASMCs）증식이급개망직단백（CRT）、CCAAT증강자결합단백α（CEBPα）표체적영향급궤제。방법（1）통과수집경불동농도CSE처리24 h적ASMCs，분위대조조、2.5%CSE조、5%CSE조、10%CSE조。이MTT법분석각조세포증식정황，채용역전록취합매련반응（RT-PCR）검측각조세포CEBPα적mRNA수평；면역인적법검측4조세포CRT、CEBPα적단백수평。（2）재10%CSE조，분별재ASMCs중전염음성대조siRNA、CRT적siRNA，비교2조ASMCs적CEBPα、CRT표체급세포증식정황。결과（1）대조조、2.5％CSE조、5％CSE조、10％CSE조ASMCs적CEBPα단백표체의차체감，CRT화세포증식정도정의차증고（P＜0.05）；이CEBPαmRNA표체차이무통계학의의（P＞0.05）。（2）재10％CSE작용하，CRTsiRNA조중ASMCs적CEBPα표체교음성대조siRNA조증강（P＜0.05），이CRT화세포증식정도균교상응음성대조siRNA조감약（P＜0.05）。결론 CSE가사ASMCs중CRT적표체증강，종이억제CEBPαmRNA적번역，사CEBPα적표체감소，최종촉진ASMCs적증식。
Objective To explore the effects and mechanism of cigarette smoke extract (CSE) on the proliferation of air?way smooth muscle cells (ASMCs) and the expression of CCAAT/enhancer-binding protein (CEBPα) and calreticulin. Meth?ods (1) The ASMCs were stimulated with different concentrations of CSE for twenty-four hours. According to the concentra?tions of CSE,the cells were divided into control group, 2.5%CSE group, 5%CSE group and 10%CSE group. The prolifera?tion of ASMCs was measured by MTT colrimetric method. The CEBPαmRNA was analyzed by RT-PCR. Western bloting as?say was performed to detect the levels of CRT and CEBPαprotein. (2) In 10%CSE group, transfection of the siRNA respec?tively for negative control or calreticulin was performed in accordance with instructions. The cell proliferation and the expres?sion of calreticulin and CEBPαwere compared in negative control siRNA group and calreticulin siRNA group. Results (1) With the increasing of the concentrations of CSE, the protein expression of CEBPαdecreased gradually (P<0.05), while the proliferation of ASMCs and the protein expression of calreticulin increased (P<0.05), but the expression of CEBPαmRNA in ASMCs showed no significant difference in groups with different concentrations of CSE (P>0.05). (2) Under the 10%CSE, the expression of CEBPαwas significantly higher in CRT siRNA group than that in negative control group (P<0.05),but the cell proliferation and CRT were significantly lower in the calreticulin siRNA group than those in negative control siRNA group (P<0.05). Conclusion The CSE exposure contributes to the expression of calreticulin protein,and then inhibits the translation of CEBPαmRNA,thus promotes the proliferation of ASMCs.
