中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
11期
773-777
,共5页
黎炳护%张莉莉%皮燕%尹延伟%杨清武%方传勤%高长越%李敬诚
黎炳護%張莉莉%皮燕%尹延偉%楊清武%方傳勤%高長越%李敬誠
려병호%장리리%피연%윤연위%양청무%방전근%고장월%리경성
Toll样受体4%增生%内皮细胞%低剪切力
Toll樣受體4%增生%內皮細胞%低剪切力
Toll양수체4%증생%내피세포%저전절력
Toll-like receptor 4%Hyperplasia%Endothelial cells,vascular%Low shear stress
目的 探讨toll样受体4(TLR4)在低剪切力(LSS)诱导的动脉内膜增生中的作用.方法 应用TLR4-/-和对照的C57 BL/6J小鼠,聚乙烯套囊制作小鼠颈动脉LSS模型,低剪切力装置处理培养的血管内皮细胞制作体外低剪切力细胞模型,常规病理切片和HE染色观察血管形态改变,Western印迹检测TLR4蛋白水平,RT-PCR检测TLR4、IL-1β、IL-6和TNF-α的mRNA表达情况,应用DNA合成率检测细胞增殖率.结果 LSS明显促进正常小鼠颈动脉内膜增生,但对TLR4-/-小鼠仅引起轻度内膜增生(42.67±16.46比7.03±2.95,P＜0.05)；LSS上调正常小鼠颈动脉中TLR4蛋白表达(2.30±0.66比0.16±0.10,P＜0.05)和促炎因子IL-1β(6.52±3.15比1.65±0.45,P＜0.01)、IL-6(16.17±7.49比6.50±1.84,P＜0.01)和TNF-α(9.98±3.77比2.72 ±1.03,P＜0.01)的mRNA表达,但对TLR4-/-小鼠仅引起IL-6和TNF-α的mRNA表达增多；LSS诱导培养的内皮细胞增殖,TLR4过表达显著增强内皮细胞的增殖能力(177±33比83±15,P＜0.05),而TLR4基因沉默则明显降低其增殖能力(40±8比83±15,P＜0.05).结论 TLR4在LSS诱导的动脉内膜增生中起重要作用；LSS可诱导TLR4的上调而使其下游炎性因子合成增加,从而促进了内皮细胞的增殖和迁移,最终导致动脉内膜增生.
目的 探討toll樣受體4(TLR4)在低剪切力(LSS)誘導的動脈內膜增生中的作用.方法 應用TLR4-/-和對照的C57 BL/6J小鼠,聚乙烯套囊製作小鼠頸動脈LSS模型,低剪切力裝置處理培養的血管內皮細胞製作體外低剪切力細胞模型,常規病理切片和HE染色觀察血管形態改變,Western印跡檢測TLR4蛋白水平,RT-PCR檢測TLR4、IL-1β、IL-6和TNF-α的mRNA錶達情況,應用DNA閤成率檢測細胞增殖率.結果 LSS明顯促進正常小鼠頸動脈內膜增生,但對TLR4-/-小鼠僅引起輕度內膜增生(42.67±16.46比7.03±2.95,P＜0.05)；LSS上調正常小鼠頸動脈中TLR4蛋白錶達(2.30±0.66比0.16±0.10,P＜0.05)和促炎因子IL-1β(6.52±3.15比1.65±0.45,P＜0.01)、IL-6(16.17±7.49比6.50±1.84,P＜0.01)和TNF-α(9.98±3.77比2.72 ±1.03,P＜0.01)的mRNA錶達,但對TLR4-/-小鼠僅引起IL-6和TNF-α的mRNA錶達增多；LSS誘導培養的內皮細胞增殖,TLR4過錶達顯著增彊內皮細胞的增殖能力(177±33比83±15,P＜0.05),而TLR4基因沉默則明顯降低其增殖能力(40±8比83±15,P＜0.05).結論 TLR4在LSS誘導的動脈內膜增生中起重要作用；LSS可誘導TLR4的上調而使其下遊炎性因子閤成增加,從而促進瞭內皮細胞的增殖和遷移,最終導緻動脈內膜增生.
목적 탐토toll양수체4(TLR4)재저전절력(LSS)유도적동맥내막증생중적작용.방법 응용TLR4-/-화대조적C57 BL/6J소서,취을희투낭제작소서경동맥LSS모형,저전절력장치처리배양적혈관내피세포제작체외저전절력세포모형,상규병리절편화HE염색관찰혈관형태개변,Western인적검측TLR4단백수평,RT-PCR검측TLR4、IL-1β、IL-6화TNF-α적mRNA표체정황,응용DNA합성솔검측세포증식솔.결과 LSS명현촉진정상소서경동맥내막증생,단대TLR4-/-소서부인기경도내막증생(42.67±16.46비7.03±2.95,P＜0.05)；LSS상조정상소서경동맥중TLR4단백표체(2.30±0.66비0.16±0.10,P＜0.05)화촉염인자IL-1β(6.52±3.15비1.65±0.45,P＜0.01)、IL-6(16.17±7.49비6.50±1.84,P＜0.01)화TNF-α(9.98±3.77비2.72 ±1.03,P＜0.01)적mRNA표체,단대TLR4-/-소서부인기IL-6화TNF-α적mRNA표체증다；LSS유도배양적내피세포증식,TLR4과표체현저증강내피세포적증식능력(177±33비83±15,P＜0.05),이TLR4기인침묵칙명현강저기증식능력(40±8비83±15,P＜0.05).결론 TLR4재LSS유도적동맥내막증생중기중요작용；LSS가유도TLR4적상조이사기하유염성인자합성증가,종이촉진료내피세포적증식화천이,최종도치동맥내막증생.
Objective To determine the role of toll like receptor 4 (TLR4) in intimal hyperplasia induced by low shear stress (LSS).Methods TLR4-/- mice and control mice C57BL/6J were used.Polyethylene cuff was placed on murine carotid to establishing a LSS model.Cultured vascular endothelial cells under LSS condition were used as an in vitro LSS cell model.Intimal hyperplasia was evaluated pathologically.TLR4 was tested by Western blot and the expression of IL-1 β,1L-6 and TNF-α mRNA were detected using RT-PCR.Cell proliferation was determined by detecting DNA synthesis.Results LSS elicited significant carotid intimal hyperplasia in normal mice but a slight neointima formation in TLR4-/- mice (42.67 ± 16.46 vs 7.03 ±2.95,P ＜0.05).LSS upregulated the expression of TLR4 (2.30 ±0.66 vs 0.16±0.10,P＜0.05),as well as the mRNA of IL-1β(6.52 ±3.15 vs 1.65 ±0.45,P＜0.01),IL-6 ( 16.17 ±7.49 vs 6.50 ± 1.84,P ＜0.01 ) and TNF-α(9.98 ±3.77 vs 2.72 ± 1.03,P ＜0.01 ) in normal mice.However,only moderate increases in IL-6 and TNF-α mRNA were observed in TLR4-/- mice.LSS induced the proliferation in cultured endothelial cells.And it was further enhanced by TLR4 overexpression (177 ±33 vs 83 ± 15,P ＜0.05) but attenuated by TLR4 silencing (40 ±8 vs 83 ± 15,P＜0.05).Conclusion TLR4 plays an important role in LSS-induced intimal hyperplasia.It is likely that LSS induces the proliferation of endothelial cells through TLR4-mediated inflammatory reaction and ultimately promotes intimal hyperplasia.
