皮下免疫热休克蛋白65对小鼠巨噬细胞胆固醇流出率的影响及机制  

Effect of subcutaneous immunization with HSP65 on cholesterol efflux from macrophagocytes in mice and its mechanism

作　　者：孙海阁[1] 沈剑刚[1] 谭迎[1] 田迪[1] 刘挺榕[1] 罗甜甜[1] 赖文岩[1] 郭志刚[1] 

机构地区：[1]南方医科大学南方医院心内科,广州510515

出　　处：《中华老年心脑血管病杂志》2014年第4期416-420,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：广东省科技计划项目(2011B031800065)

摘　　要：目的观察皮下免疫不同剂量热休克蛋白65(HSP65)对载脂蛋白E缺失小鼠动脉粥样硬化及巨噬细胞TC流出率的影响。方法选择apoE-/-小鼠24只,随机分为磷酸盐缓冲液(PBS)对照组、5μg HSP65组及25μg HSP65组,每组8只,第3、6周进行皮下免疫。实验第16周末取标本,分离小鼠主动脉测定血管斑块面积;用自动生化仪测定小鼠血脂水平,酶联免疫吸附法分别检测血清HSP65抗体滴度及炎性因子白细胞介素10(IL-10)和干扰素γ水平,液闪计数仪检测巨噬细胞TC流出率;提取肝脏mRNA及膜蛋白,分别检测ATP结合盒转运蛋白A1(ABCA1)和G1(ABCG1)、B族Ⅰ型清道夫受体(SR-BⅠ)的表达。结果与PBS对照组比较,5μg HSP65组和25μg HSP65组TC流出率明显降低,HSP65抗体滴度和干扰素γ水平明显升高,主动脉粥样硬化斑块面积明显增加(P<0.05,P<0.01);其中25μg HSP65组血清HDL-C和IL-10表达较PBS对照组明显降低(P<0.01);5μg HSP65组和25μg HSP65组ABCA1、ABCG1及SR-BⅠmRNA表达明显低于PBS对照组(P<0.05)。结论皮下免疫HSP65可抑制小鼠巨噬细胞TC流出,可能是通过下调ABCA1、ABCG1和SR-BⅠ的表达。Objective To study the effect of subcutaneous immunization with HSP65 on atheroscle- rosis and cholesterol efflux from macrophagocytes in apoE/mice and its mechanism. Methods Twenty-four apoE/mice aged 8 weeks were randomly divided into PBS group （control group）,5 μg HSP65 group and 25 μg HSP65 group （8 ineach group）. The animals underwent subcutaneous immunization with HSP65 at weeks 3 and 6. Samples were taken at week 16. Area of atorta plaques,serum lipid level, serum HSP65 antibody titer, and IL-10 and interferon γ level were measured by ELISA. Cholesterol efflux was assayed by liquid scintillation spectrometry. Expres- sions of ABCA1,SR-B I and ABCG1 mRNA and protein were detected by RT-PCR and Western blot,respectively. Results The cholesterol efflux was significantly lower,the serum HSP65 anti- body titer and interferong level were significantly higher, the area of atorta plaques was signifi- cantly larger in 5 μg HSP65 group and 25 μg HSP65 group than in control group （P〈0.05,P〈 0.01）. The expression levels of HDL-C,IL-10,and ABCA1 ,ABCG1 and SR-B I mRNA and pro- tein were significantly lower in 5 μg HSP65 group and 25 μg HSP65 group than in control group （P〈0.01）. Conclusion Subcutaneous immunization with HSP65 inhibits cholesterol efflux from macrophagocytes in mice by down-regulating the expression of ABCA1, ABCG1 and SR-B I .

关 键 词：热休克蛋白质类 载脂蛋白E类 巨噬细胞 动脉粥样硬化 白细胞介素10 干扰素Ⅱ型 

分 类 号：R543.5[医药卫生—心血管疾病]