摘要
目的探讨过氧化物酶体增殖物激活受体α亚型(PPARα)激动剂非诺贝特对人脑微血管内皮细胞(BMEC)与小鼠脑微血管组织细胞外铜/锌超氧化物歧化酶(SOD3)表达的调节作用及其机制。方法将培养好的BMEC随机分为两部分,分别加入终浓度为10μg/m L非诺贝特、同等体积的DMSO处理12 h;将15只小鼠随机分为两部分,分别给予30 mg/kg非诺贝特、10%的羧甲基纤维素混悬液10 m L/kg连续灌胃7 d,处死小鼠取脑组织,提取脑微血管;用RT-PCR技术检测BMEC与脑微血管组织内过氧化物酶体增殖物激活受体α亚型(PPARα)、SOD3的mRNA表达。构建含SOD3启动子荧光素酶报告质粒p GL4 m SOD3,用突变试剂盒定点获得过氧化物酶体增殖物反应元件(PPRE)序列突变的荧光素酶报告质粒p GL4 mu m SOD3,将p GL4 m SOD3、p GL4 mu m SOD3转染BMEC 24 h,取部分细胞用终浓度10μg/m L的非诺贝特处理12 h,检测细胞荧光素酶活性。结果经非诺贝特处理后,BMEC及小鼠脑微血管组织中PPARα、SOD3 mRNA表达均升高(P均<0.05)。转染p GL4 m SOD3 24 h,经非诺贝特处理后BMEC内p GL4 m SOD3的荧光素酶活性增强(P<0.05);转染p GL4 mu m SOD3 24 h,经非诺贝特处理的BMEC荧光素酶活性变化不明显(P>0.05)。结论非诺贝特通过激活PPARα从而调节人BMEC与脑微血管组织中SOD3的表达。
Objective To investigate the regulating effect of fenofibrate,the agonist of peroxisome proliferator-activated receptor α（ PPARα）,on the superoxide dismutase 3（ SOD3） expression in human brain microvascular endothelial cells（ BMECs） and mouse brain microvascular tissues as well as the possible mechanism. Methods The cultured BMECs were randomly divided into two groups,which were seperately treated with 10 μg / ml of fenofibrate and the same volume of DMSO for 12 h. Fifteen mice were divided into two groups,which were treated with 30 mg / kg fenofibrate and 10 m L / kg of 10% CMC by gavage for 7 days. The tRNA was extracted and the brain microvessel was isolated. The mRNA expression levels of PPARα and SOD3 were detected by RT-PCR. We constructed a luciferase reporter plasmid p GL4 m SOD3 containing SOD3 promoter. The luciferase reporter plasmid p GL4 mu m SOD3 mutated from PPRE sequence were obtained by using the mutagenesis kit. The BMECs were transfected with p GL4 m SOD3 and p GL4 mu m SOD3 for 24 h. The cells were treated with fenofibrate at a final concentration of 10 μg / m L for 12 h in order to measure the luciferase activity. Results The mRNA expression of PPARα and SOD3 was increased in the BMECs and brain microvessel after the treatment of fenofibrate（ all P 0. 05）. The luciferase activity of p GL4 m SOD3 in BMECs treated with fenofibrate was enhanced after transfection of p GL4 m SOD3 for 24 h（ P 0. 05）. The luciferase activity of BMECs treated with fenofibrate did not significantly change after transfection of p GL4 mu m SOD3 for 24 h（ P 0. 05）. Conclusion The SOD3 expression in the human BMECs and mouse brain microvascular tissues is regulated by fenofibrate through activating PPARα.
作者
杜小珊
杨锡彤
徐弘扬
程建杰
王光明
DU Xiaoshan YANG Xitong XU Hongyang CHEN Jianjie WANG Guangming(The First Affiliated Hospital of Dali University, Dali 671000, China)
出处
《山东医药》
CAS
北大核心
2017年第5期4-7,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81360206)
云南省中青年学术技术带头人后备人才基金项目(2014HB025)
关键词
非诺贝特
细胞外铜/锌超氧化物歧化酶
脑微血管
脑微血管内皮细胞
fenofibrate
extracellular Cu / Zn superoxide dismutase
brain microvascular
brain microvascular endothelial cells
