非诺贝特对人BMEC与小鼠脑微血管组织SOD3表达的影响及机制

发布时间：2018-01-24 13:09

本文关键词： 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　出处：《山东医药》2017年05期 　论文类型：期刊论文

【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(P0.05);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(P0.05)。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。
[Abstract]:Objective to investigate the effects of fenofibrate, a peroxisome proliferator activated receptor 伪 (PPAR 伪) agonist, on human microvascular endothelial cells (BMECs). The regulation and mechanism of extracellular copper / zinc superoxide dismutase (SOD3) expression in rat brain microvessels were studied. Methods cultured BMEC was randomly divided into two parts. The final concentration of fenofibrate was 10 渭 g / mL and treated with the same volume of DMSO for 12 h. Fifteen mice were randomly divided into two groups. The mice were given 10% carboxymethyl cellulose suspension (10 mL / kg) for 30 mg/kg for 7 days. Extraction of brain microvessels; RT-PCR technique was used to detect BMEC and peroxisome proliferator activated receptor 伪 (PPAR- 伪) in brain microvascular tissue. MRNA expression of SOD3. Construction of SOD3 promoter luciferase reporter plasmid p GL4 m SOD3. The mutated luciferase reporter plasmid p GL4 mu m SOD3 of peroxisome proliferator response element (PPRE) was obtained by using mutation kit. After transfection of p GL4 m SOD 3 + p GL4 mu m SOD3 into BMEC for 24 h, some cells were treated with fenofibrate at the final concentration of 10 渭 g / mL for 12 h. Results after fenofibrate treatment, BMEC and PPAR 伪 in mouse brain microvessels were detected. The expression of SOD3 mRNA was increased (P < 0.05) and p GL4 m SOD3 was transfected for 24 h. The luciferase activity of p GL4 m SOD3 in BMEC was enhanced by fenofibrate treatment (P0. 05). Transfection of p GL4 mu m SOD3 for 24 h. The change of luciferase activity of BMEC treated with fenofibrate was not obvious (P0.05). Conclusion fenofibrate regulates the expression of SOD3 in human BMEC and brain microvessels by activating PPAR 伪.
【作者单位】：大理大学第一附属医院;
【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)
【分类号】：R743
【正文快照】： 超氧化物歧化酶(SOD)在机体中能消除新陈代谢过程中产生的有害物质,其有三种亚型:胞质内的SOD1、线粒体内SOD2及细胞外铜/锌超氧化物歧化酶(SOD3)[1]。SOD的耗竭或内源性抗氧化系统的破坏是缺氧导致组织损伤的机制[2,3]。缺血会导致脑组织中SOD表达下降。过氧化物酶体增殖物激

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