高糖环境中雄激素受体对心肌成纤维细胞增殖和脂质合成能力的影响  

Effects of AR on proliferation and lipid synthesis of myocardial fibroblasts in high glucose environment

作　　者：温馨 周洋 林丽婵 刘芷言 宋凯 陶辉[4] 臧洪梅[1] WEN Xin;ZHOU Yang;LIN Li-chan;LIU Zhi-yan;SONG Kai;TAO Hui;ZANG Hong-mei(School of Pharmacy,Anhui Medical University,Hefei 230032,China;Dept of Pharmacology,the Third Affiliated Hospital of Anhui Medical University,Hefei First People′s Hospital,Hefei 230061,China;Dept of Cardiothoracic Surgery,the Second Affiliated Hospital of Anhui Medical University,Hefei 230601,China;Dept of Anesthesiology,the Second Affiliated Hospital of Anhui Medical University,Hefei 230601,China)

机构地区：[1]安徽医科大学药学院,安徽合肥230032 [2]安徽医科大学第三附属医院药学部,合肥市第一人民医院,安徽合肥230061 [3]安徽医科大学第二附属医院心胸外科,安徽合肥230601 [4]安徽医科大学第二附属医院麻醉科,安徽合肥230601

出　　处：《中国药理学通报》2023年第12期2274-2280,共7页Chinese Pharmacological Bulletin

基　　金：安徽省教育厅高校自然科学基金重大项目(No 2023AH040090);安徽省转化医学研究基金项目(No 2021zhyx-C61)。

摘　　要：目的探究雄激素受体(androgen receptor,AR)对高糖环境下心肌成纤维细胞(cardiac fibroblasts,CFs)增殖和脂质合成能力的影响及可能的分子机制。方法解剖新生乳鼠的心脏进行CFs原代培养,镜下观察CFs生长状态,并通过波形蛋白的免疫荧光鉴定细胞属性。细胞贴壁后分为空白对照组、高糖模型组、阴性对照组,过表达AR组,除空白对照组葡萄糖浓度为5.5 mmol·L^(-1)外,其余各组葡萄糖浓度均为33.0 mmol·L^(-1)。培养24 h后通过Western blot和RT-qPCR检测AR、脂肪酸合成酶(fatty acid synthase,FASN)、PCNA、cyclin D1、α-SMA、collagenⅠ的表达情况。油红O和CCK-8分别检测脂质合成和细胞增殖能力的改变。结果与空白对照组相比,高糖模型组中CFs的脂质合成和增殖能力增强。Western blot和RT-qPCR结果显示AR的蛋白和mRNA的表达降低,而FASN、增殖标志蛋白PCNA、cyclin D1和纤维化标志蛋白α-SMA、collagenⅠ的蛋白和mRNA的表达增加。在过表达AR质粒载体转染高糖模型组CFs后,过表达组CFs中的FASN、PCNA、cyclin D1、α-SMA、collagenⅠ蛋白和mRNA同空载体组相比表达下降。同时,油红O染色和CCK-8结果分别表明过表达AR组的脂质合成和增殖能力相比空载体组减弱。结论高糖环境下心肌成纤维细胞的增殖和脂质合成能力增强,其作用机制可能AR调控脂质合成有关。Aim To explore the effect of androgen receptor AR on the proliferation and lipid synthesis of cardiac fibroblasts under high-glucose conditions and the possible molecular mechanism.Methods The hearts of neonatal rats were dissected for primary culture of cardiac fibroblasts.Then the growth status of CFs was observed under the inverted microscope,and the identification of CFs was performed by immunofluorescence staining using anti-vimentin.After cell adherence,the cells were divided into blank control group,high glucose model group,negative control group,and overexpressed AR group.The glucose concentration was 33.0 mmol·L^(-1)except that the blank control group was 5.5 mmol·L^(-1).After 24 hours of CFs culture,Western blot and RT-qPCR were used to detect the expression of AR,FASN,PCNA,cyclin D1,α-SMA,and collagenⅠ.Oil red O and CCK-8 were used to detect the changes in lipid synthesis and cell proliferation ability,respectively.Results Compared with the blank control group,the lipid synthesis and proliferation of CFs in the high glucose model group were enhanced.Western blot and RT-qPCR results showed that the expression of AR decreased,while the expression of fat lipid synthase(FASN),proliferation marker PCNA,cyclin D1 and fibrosis markerα-SMA and collagenⅠincreased.After AR overexpressed plasmid was transfected into the CFs treated by high glucose,AR overexpression markedly decreased the expression of FASN,PCNA,cyclin D1,α-SMA and collagenⅠcompared with the empty plasmid‐transfected group.Meanwhile,oil red O staining and CCK-8 results showed that the lipid synthesis and proliferation ability of the overexpressed AR group decreased compared with the empty vector group,respectively.Conclusions High glucose promotes the proliferation and lipid synthesis of cardiac fibroblasts.Besides,the mechanism may be related to the regulation of lipid synthesis regulated by AR.

关 键 词：脂质合成 心肌纤维化 细胞增殖 心肌成纤维细胞 细胞代谢 雄激素受体 

分 类 号：R-332[医药卫生] R322.11R329.28R344R392.11