摘要
目的探讨移植转染血红素氧合酶1(HO-1)基因的巨噬细胞在减轻大鼠急性心肌梗死(AMI)区炎症和氧化应激损伤中的作用与机制。方法体外培养大鼠肺泡巨噬细胞(Mφ)并分为对照组、腺病毒(Ad)空载体转染巨噬细胞组(Ad-Mφ组)、含HO-1基因的Ad转染巨噬细胞组(Ad-HO-1-Mφ组)。检测各组的细胞增殖活力、HO-1蛋白表达量及培养液中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-10等细胞因子表达量。另以冠状动脉结扎法建立SD大鼠AMI模型并随机分为对照组、Ad-Mφ移植组和Ad-HO-1-Mφ移植组,分别于心肌梗死区多点注射磷酸缓冲液(PBS)、含Ad-Mφ(2×109/L)的PBS液、含Ad-HO-1-Mφ(2×109/L)的PBS液各100μL,5天后处死各组大鼠,取梗死区心肌组织检测HO-1蛋白及IL-12、TNF-α、IL-10、单核细胞趋化蛋白1(MCP-1)等表达量,同时检测总抗氧化能力(T-AOC)和丙二醛(MDA)含量。结果腺病毒转染本身不影响巨噬细胞活力,但却能将HO-1基因转入巨噬细胞并获得高效表达(P〈0.05),转染HO-1基因的巨噬细胞可明显降低促炎因子TNF-α、IL-6表达并升高抗炎因子IL-10的表达(均为P〈0.05)。与对照组及Ad-Mφ移植组比较,Ad-HO-1-Mφ移植组心肌梗死区HO-1蛋白表达明显升高(P〈0.05),促炎因子TNF-α、IL-12、MCP-1表达减少而抗炎因子IL-10表达增加,对应的T-AOC升高而MDA显著降低(均为P〈0.05)。结论 Ad-HO-1-Mφ能够高表达HO-1蛋白并向具有抗炎特性的M2型极化。将Ad-HO-1-Mφ移植到大鼠急性心肌梗死区仍能高表达HO-1蛋白并显著减轻梗死区的炎症和氧化应激水平。
Aim To explore the effects and mechanisms of transplantation of macrophages transfected with heme oxygenase-1( HO-1) gene by adenovirus vectors on inflammation,oxidative stress in acute myocardial infarction( AMI) area. Methods Rat alveolar macrophages were cultured in vitro and divided into three groups and treated differently:control group,the cells were not transfected with adenovirus; Ad-Mφ group,the cells were transfected with blank adenovirus vector; Ad-HO-1-Mφ group,the cells were transfected with adenovirus carrying the HO-1 gene. The proliferative activity of macrophages and the expression of HO-1 protein as well as the inflammatory cytokines such as tumor necrosis factor( TNF-α),interleukelin-6( IL-6),IL-10 in the supernatant were detected. In addition,The AMI model in rats were established by ligating coronary artery and randomly divided into three groups: the simple AMI group,the Ad-Mφ transplantation group and the Ad-HO-1-Mφ transplantation group. PBS,PBS containing 2×105 Ad-Mφ and PBS containing 2×105 Ad-HO-1-Mφ were injected into the myocardial infarction area of rats in each group respectively. 5 days later,the expression of HO-1 protein and the inflammatory cytokines such as IL-12,TNF-α,IL-10,monocyte chemotactic protein 1( MCP-1) as well as the total antioxidant capacity( T-AOC) and malondialdehyde( MDA) content in infarcted myocardium were detected. Results Adenovirus transfection itself did not affect macrophage proliferation activity,but could transfer HO-1 gene into macrophage and obtain high expression. Ad-HO-1-Mφ could significantly reduce the expression of pro-inflammatory cytokine TNF-α,IL-6 and increase the expression of anti-inflammatory factor IL-10( P〈0. 05). Compared with the simple AMI group and the Ad-Mφ transplantation group,the infarcted area of Ad-HO-1-Mφ transplantation group showed much more expression of HO-1 protein and IL-10 and less expression of TNF-α,IL-12,MCP-1 while T-AOC increased and MDA content decreased significantly( P〈0. 05 respectively). Conclusion Ad-HO-1-Mφ could overexpress HO-1 protein and polarize into M2 phenotype which has powerful anti-inflammatory properties. After being transplanted into myocardial infarcted area,the Ad-HO-1-Mφ could still overexpress HO-1 protein and significantly reduce the inflammation and oxidative stress levels in myocardial infracted area.
作者
李谌
黄维义
LI Chen;HUANG Wei-Yi(The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, China;The People's Hospital of Yubei District of Chongqing , Chongqing 401120, China)
出处
《中国动脉硬化杂志》
CAS
2018年第5期467-473,共7页
Chinese Journal of Arteriosclerosis
基金
泸州市科技局项目(2013LZLY-J15)
关键词
腺病毒转染
巨噬细胞
血红素氧合酶1
急性心肌梗死
炎症和氧化应激
Adenovirus transfection
Macrophages
Heme oxygenase-1
Acute myocardial infarction
Inflammation and oxidative stress