罗格列酮预处理对凝血酶激活的小胶质细胞PPARγ、Nrf2及HO-1表达的影响  

Rosiglitazone pretreatment influences expression of PPARγ,Nrf2 and HO-1 in thrombin-activated microglia

作　　者：杭航[1] 王丽琨[1] 伍国锋[1] 陈星宇[1] 

机构地区：[1]贵州医科大学附属医院急诊医学科/急诊医学教研室,贵州贵阳550004

出　　处：《中国病理生理杂志》2016年第4期671-679,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81460185/H09106);贵州省科技基金资助项目(No.2013-2043)

摘　　要：目的:采用凝血酶激活新生大鼠神经胶质细胞,观察罗格列酮预处理对小胶质细胞过氧化物酶体增殖物活化受体γ(PPARγ)、核因子E2相关因子2(Nrf-2)及血红素加氧酶-1(HO-1)表达的影响。方法:用新生SD大鼠的脑组织,体外培养原代小胶质细胞14 d左右分离收集细胞,分为:正常对照组、凝血酶刺激组、罗格列酮干预组(罗格列酮+凝血酶组)和维甲酸干预组(维甲酸+凝血酶组)进行实验。分别采用免疫组化染色、real-time PCR和Western blot检测PPARγ、Nrf2和HO-1的表达并进行统计分析。结果:免疫组化染色显示,与对照组比较,刺激组、罗格列酮+凝血酶组及维甲酸+凝血酶组的PPARγ、Nrf2和HO-1染色细胞数均增多。Real-time PCR结果显示罗格列酮+凝血酶组PPARγ、Nrf2及HO-1的mRNA表达均显著高于刺激组、对照组及维甲酸+凝血酶组(P<0.01),维甲酸+凝血酶组Nrf2及HO-1的mRNA表达均较刺激组和罗格列酮+凝血酶组降低(P<0.01)。Western blot结果显示,罗格列酮+凝血酶组PPARγ、Nrf2及HO-1的蛋白表达也明显高于刺激组、对照组及维甲酸+凝血酶组(P<0.01),维甲酸+凝血酶组Nrf2及HO-1的蛋白表达均较刺激组和罗格列酮+凝血酶组降低(P<0.01)。结论:罗格列酮预处理后可增加凝血酶激活的小胶质细胞PPARγ、Nrf2及HO-1的表达,通过维甲酸预处理抑制Nrf2的表达后,其下游基因HO-1表达也受影响,说明PPARγ抗氧化作用可能是通过Nrf2调控下游基因实现的。AIM： To observe the effect of rosiglitazone（ RGZ） pretreatment on the expression of peroxisome proliferator-activated receptor γ（ PPARγ）,nuclear factor E2-related factor 2（ Nrf2） and heme oxygenase-1（ HO-1） in the microglia cells activated by thrombin. METHODS： Microglia cells were obtained from the brain tissues of the newborn rats and were primarily cultured in vitro. After cultured for 14 d,the microglia cells were used in the experiment. The isolated microglia cells were randomly divided into normal control group,thrombin stimulation group（ TH group）,rosiglitazone intervention group（ RGZ ＋ TH group） and retinoic acid intervention group（ RA ＋ TH group）. The expression of PPARγ,Nrf2 and HO-1 was observed by immunocytochemistry,real-time PCR and Western blot. RESULTS： The number of positive staining cells of PPARγ,Nrf2 and HO-1 in TH group,RGZ ＋ TH group and RA ＋ TH group were increased remarkably as compared with control group. The significant increases in PPARγ,Nrf2 and HO-1 were observed in RGZ ＋ TH group compared with other groups. The mRNA expression of PPARγ,Nrf2 and HO-1 in RGZ ＋ TH group was increased significantly as compared with TH group,control group or RA ＋ TH group（ P 0. 01）,Besides,the mRNA expression of Nrf2 and HO-1 in RA ＋ TH group was decreased as compared with TH group or RGZ ＋ TH group（ P 0. 01）. The protein levels of PPARγ,Nrf2 and HO-1 in RGZ ＋ TH group were significantly increased as compared with TH group,control group or RA ＋ TH group（ P 0. 01）. The protein expression of Nrf2 and HO-1 in RA ＋ TH group was decreased as compared with TH group or RGZ ＋ TH group（ P 0. 01）. CONCLUSION： Rosiglitazone pretreatment might increase the expression of PPARγ,Nrf2 and HO-1 in the microglia cells activated by thrombin. By inhibiting the expression of Nrf2 after RA pretreatment,the expression of the downstream gene HO-1 is also influenced. The anti-oxidative stress effects of rosiglitazone might be achieved partly by

关 键 词：小胶质细胞 脑出血 罗格列酮 过氧化物酶体增殖物活化受体Γ 核因子E2相关因子2 血红素加氧酶-1 

分 类 号：R459.7[医药卫生—急诊医学]