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目的探讨脂筏在柯萨奇B3病毒(CVB3)感染心肌细胞中的作用,旨在为阐明CVB3致病的分子机理及寻找新的抗病毒靶点提供依据。方法用甲基-β-环糊精(MβCD)去除细胞膜维持脂筏稳定性的胆固醇分子后感染CVB3,用Western blot法检测CVB3 VP1蛋白的表达并测定病毒的滴度,同时观察补充外源性胆固醇恢复MβCD对CVB3感染的抑制作用。结果用MβCD去除胆固醇分子破坏细胞膜脂筏结构,可抑制CVB3感染细胞;1.2、5.5和10 mmol/L MβCD去除细胞膜胆固醇CVB3滴度分别为(4.2±0.06)、(3.5±1.05)、(3.0±0.15)和(2.0±0.15)lgP-FU/mL,均低于无MβCD处理对照组的(5.7±0.06)lgPFU/mL(P<0.001);补充外源性胆固醇可恢复MβCD对CVB3感染的抑制作用。结论细胞膜脂筏在CVB3感染心肌细胞中起重要作用,是病毒进入细胞的关键因素。
Objective To investigate the role of lipid raft in the infection of Coxsackievirus B3 (CVB3) in cardiomyocytes and to provide a basis for elucidating the molecular mechanism of CVB3 pathogenesis and searching for new anti-viral targets. Methods CVB3 was infected by removing cholesterol molecules that retained lipid raft stability by methyl β -cyclodextrin (MβCD). The expression of CVB3 VP1 protein was detected by Western blot and the titer of the virus was assayed. The exogenous Cholesterol Restores the Inhibitory Effect of MβCD on CVB3 Infection. Results Removal of cholesterol molecules by MβCD destroied the lipid raft structure of the cell membrane and inhibited CVB3-infected cells. The titers of CVB3 at the cell membrane cholesterol levels of 1.2, 5.5 and 10 mmol / L MβCD were (4.2 ± 0.06), (3.5 ± 1.05) and (3.0 ± 0.15) and (2.0 ± 0.15) lgP-FU / mL, respectively, were lower than those in the control group without MβCD (5.7 ± 0.06) lgPFU / mL effect. Conclusions Membrane lipid rafts play an important role in CVB3-infected cardiomyocytes and are the key factors of virus entering cells.