CYP19增强脱氢表雄酮抑制高脂诱导的兔主动脉及人脐静脉内皮细胞MMP-9的表达

基础医学与临床 ›› 2019, Vol. 39 ›› Issue (1): 53-58.

CYP19增强脱氢表雄酮抑制高脂诱导的兔主动脉及人脐静脉内皮细胞MMP-9的表达

周颖,赵敏,肖芳,李桃,李晓超

秦皇岛市第一医院

收稿日期:2018-02-25 修回日期:2018-05-30 出版日期:2019-01-05 发布日期:2018-12-28
通讯作者: 赵敏 E-mail:496437510@qq.com
基金资助:
秦皇岛市科技支撑计划项目

CYP19 enhances inhibition of dehydroepiandrosterone on high fat-induced MMP-9 expression in rabbit aorta and HUVEC

Received:2018-02-25 Revised:2018-05-30 Online:2019-01-05 Published:2018-12-28
Contact: Min ZHAO E-mail:496437510@qq.com

摘要/Abstract

摘要： 目的探讨脱氢表雄酮（DHEA）抗动脉粥样硬化（AS）的作用及其机制。方法将人脐静脉内皮细胞（HUVEC）分为对照组、ox-LDL组（30 mg/L ox-LDL）、DHEA（低和高浓度）干预组（30 mg/L ox-LDL + 0.1和1 μmol/L DHEA）、DHEA＋全反式维甲酸（ATRA）干预组（30 mg/L ox-LDL + 1 μmol/L DHEA+ 0.01 μmol/L ATRA）及DHEA组（1 μmol/L）与上一行DHEA干预组重复？答：DHEA组是只加入DHEA作用组，DHEA干预组是同时加入ox-LDL和DHEA共同作用组，二者不重复。用real-time PCR和ELISA检测MMP-9 mRNA及蛋白的表达。通过脂质体介导的方法将pcDNA3.1-CYP19-GFP真核表达质粒（细胞色素P450芳香酶基因）及pcDNA3.1-GFP空质粒分别转染至人脐静脉内皮细胞（HUVEC），给予ox-LDL诱导及DHEA干预，用real-time PCR和ELISA检测转染细胞MMP-9 mRNA及蛋白的表达。将兔分为对照组、高脂组（口服1％胆固醇和3％猪油）、DHEA干预组（口服0.27% DHEA）、DHEA＋ATRA干预组（口服0.6 mg/kg﹒d ATRA）及单DHEA组。用real-time PCR和免疫组织化学法检测兔主动脉MMP-9 mRNA及蛋白的表达。结果 ox-LDL组HUVEC MMP-9的表达较对照组明显升高（P<0.05）；DHEA干预组呈浓度依赖性使MMP-9的表达明显回降（P<0.05）。高脂组兔主动脉MMP-9的表达较对照组明显升高（P<0.05）；DHEA干预组能明显缓解高脂组的变化（P<0.05）。CYP19+ox-LDL+DHEA组较空质粒+ox-LDL+DHEA组MMP-9的表达显著降低（P<0.05）。结论 DHEA能抑制高脂诱导的兔主动脉及HUVEC MMP-9的表达，过表达细胞色素P450芳香酶基因（CYP19）能增强此作用。

关键词: 动脉粥样硬化, 脱氢表雄酮, 细胞色素P450芳香酶基因, 基质金属蛋白酶-9, 人脐静脉内皮细胞

Abstract: Objective To investigate the effect of DHEA on antiatherosclerosis and its mechanisms. Methods In vitro cultured HUVEC were divided into 6 groups: control group; ox-LDL group (30 mg/L ox-LDL); DHEA (low and high concentration) intervention group (30 mg/L ox-LDL + 0.1 and 1 μmol/L DHEA); DHEA＋ATRA intervention group (30 mg/L ox-LDL + 1 μmol/L DHEA + 0.01 μmol/L ATRA); DHEA group (1 μmol/L). The expressions of HUVEC MMP-9 mRNA and protein were determined by RT-qPCR and ELISA respectively. The eukaryotic expression plasmid pcDNA3.1-CYP19-GFP (CYP19) and pcDNA3.1-GFP were transfected into HUVEC respectively. The transfected HUVEC were treated with ox-LDL and DHEA. The expressions of MMP-9 mRNA and protein of transfection groups were determined by RT-qPCR and ELISA. Rabbits were divided into 5 groups: control group; high lipid group (oral 1％cholesterol and 3％lard oil); DHEA intervention group (oral 0.27% DHEA); DHEA＋ATRA intervention group (oral 0.6 mg/kg?d ATRA); DHEA group. The expressions of aortic MMP-9 mRNA and protein were determined by RT-qPCR and immunohistochemistry. Results The expression of MMP-9 in HUVEC in ox-LDL group was significantly increased as compared with control group (P<0.05); The expressions of MMP-9 in DHEA (low and high concentration) intervention groups were obviously decreased in a dose-dependent manner (P<0.05). The expression of aortic MMP-9 in high lipid group was significantly increased as compared with control group (P<0.05). Compared with high lipid group the expression of MMP-9 in DHEA intervention group was obviously decreased (P<0.05). The expression of MMP-9 in CYP19+ox-LDL+DHEA group was significantly decreased as compared with empty plasmid+ox-LDL+DHEA group (P<0.05). Conclusions DHEA inhibits high lipid induced MMP-9 expression in rabbit aorta and HUVEC. CYP19 over-expression can enhance the effect of DHEA.

Key words: artherosclerosis, dehydroepiandrosterone, CYP19, matrix metalloproteinase-9, human umbilical venous endothelial cells

周颖赵敏肖芳李桃李晓超. CYP19增强脱氢表雄酮抑制高脂诱导的兔主动脉及人脐静脉内皮细胞MMP-9的表达[J]. 基础医学与临床, 2019, 39(1): 53-58.

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