DcR3重组蛋白对糖尿病大鼠心肌组织凋亡相关分子的表达及细胞凋亡的作用  

Action of DcR3 recombinant protein of myocardial tissue in diabetic rats

作　　者：黄黎月[1] 庄国洪[2] 丘劲华[2] 陈福[3] 陶惠然[2] 

机构地区：[1]厦门医学高等专科学校,厦门361008 [2]厦门大学医学院,厦门361102 [3]厦门大学医学院实验动物中心,厦门361102

出　　处：《中国免疫学杂志》2015年第4期472-476,共5页Chinese Journal of Immunology

摘　　要：目的:研究诱骗受体3(DcR3)在正常大鼠、糖尿病(DM)大鼠心肌组织的表达,DcR3重组蛋白对心肌组织凋亡相关分子表达以及心肌细胞凋亡的影响,探讨DcR3对DM大鼠心肌细胞凋亡的作用。方法:一次性腹腔注射链脲佐菌素建立大鼠DM模型,尾静脉注射不同剂量的DcR3重组蛋白[1.2 mg/(鼠·d)、0.8 mg/(鼠·d)、0.4 mg/(鼠·d)]40 d。RTPCR检测心肌组织DcR3 mRNA、Fas mRNA、FasL mRNA的表达。Wester blot分析凋亡相关蛋白Bcl-2、Caspase-8的表达。双抗夹心ELISA检测血液中IL-1β、TNF-α及LFN-γ水平的变化,HE染色观察心肌细胞凋亡的百分率。结果:DcR3治疗组大鼠与DM组比较心肌组织DcR3 mRNA高表达,Fas mRNA、FasL mRNA表达下调。Caspase-8蛋白水平下调,Bcl-2蛋白水平上调,以中剂量组的作用最明显。各DcR3治疗组血清IL-1β、TNF-α和IFN-γ水平均有不同程度的降低(P<0.05,P<0.01)。心肌细胞凋亡的百分率下降(P<0.05)。结论:DcR3重组蛋白有抑制DM大鼠心肌细胞凋亡的作用,其机制与竞争Fas,阻断FasL诱导细胞凋亡,心肌细胞表达DcR3,凋亡相关因子Caspase-8下调、Bcl-2上调及细胞因子水平的降低有关。Objective：To study the expression of DcR3 of myocardial tissue in diabetic mouse and normal rats and the impact of DcR3 recombinant protein to the expression of related molecules and myocardial cell apoptosis to discuss the action of DcR3 to myocardial cell apoptosis in Diabetic rats. Methods： Intraperitoneally injected streptozotocin one time to establish the model of Diabetic rats. Injected different doses of DcR3 recombinant protein to tail vein[1. 2 mg /（ rat·d）,0. 8 mg /（ rat·d）,0. 4 mg /（ rat·d）]40 d.The expression of DcR3 mRNA,Fas mRNA and Fas L mRNA of myocardial tissue was detected with RT-PCR;the expression of apoptosis related molecules Bcl-2 and Caspase-8 was analyzed with Western blot;the IL-1β,TNF-α and LFN-γ of the blood was detected with double antibody sandwich ELISA;the percentage of myocardial cell apoptosis was observed with HE dyeing. Results： To compare the DcR3 treatment group with diabetic group,the expression DcR3 of myocardial tissue was high,the expression of Fas mRNA and Fas L mRNA was descended. The Caspase-8 protein was ascended and the Bcl-2 protein was descended. The middle dose group was the most obvious. the IL-1β,TNF-α and IFN-γ in the blood was descended differently in each DcR3 treatment group（ P〈0. 05,P〈0. 01）. The percentage of myocardial cell apoptosis was declined（ P〈0. 05）. Conclusion： DcR3 recombinant protein have the action of inhibiting the rats＇ myocardial cell apoptosis,the mechanism is related to competing with Fas,blocking-up Fas L of inducing apoptosis,expressing DcR3 of myocardial cell,the descending of apoptosis related factors Caspase-8,the ascending of Bcl-2 and the reduction of cytokine levels.

关 键 词：DcR3重组蛋白 糖尿病 心肌细胞凋亡 凋亡相关分子 

分 类 号：R392.6[医药卫生—免疫学]