摘要
目的:探讨达格列净对2型糖尿病小鼠胸主动脉内皮细胞损伤及高糖诱导的人脐静脉内皮细胞(HUVECs)损伤的抑制作用及其分子机制。方法:(1)将20只2型糖尿病db/db小鼠随机分为模型组和达格列净组,另取10只正常db/m小鼠作为对照组。饲养40 d后麻醉处死,迅速摘取眼球取血,分离血清,并收集胸主动脉组织。HE染色观察胸主动脉组织形态学改变,Western blot法检测小鼠胸主动脉组织中ATP合酶α亚基(ATP5A1)、内皮素1(ET-1)和前列环素(PGI_(2))蛋白表达水平。(2)将HUVECs分为对照组、DMSO组、高糖组和高糖+达格列净组;采用活性氧(ROS)荧光探针和线粒体ATP荧光探针检测HUVECs中ROS及线粒体ATP荧光强度;采用免疫细胞化学和Western blot检测HUVECs中ATP5A1、ET-1和PGI_(2)蛋白表达水平。结果:达格列净可显著减轻db/db小鼠胸主动脉组织损伤;与高糖组相比,达格列净治疗后HUVECs中ROS荧光强度显著降低,线粒体ATP荧光强度显著升高(均P<0.01);免疫细胞化学和Western blot结果显示,达格列净治疗后ATP5A1和PGI_(2)蛋白表达水平显著升高,ET-1蛋白表达水平显著降低(均P<0.01);ATP5A1过表达质粒转染后,HUVECs中ATP5A1和PGI_(2)蛋白表达水平显著升高,ET-1表达水平显著降低(均P<0.01)。结论:达格列净减轻2型糖尿病小鼠胸主动脉内皮细胞损伤及高糖诱导的HUVECs损伤,其机制可能与调控ATP5A1表达、改善线粒体功能有关。
AIM To investigate the inhitory effects of dapagliflozin on thoracic aortic endothelial cell injury in type 2 diabetic mice and high glucose-induced injury of human umbilical vein endothelial cells(HUVECs),and to explore the related molecular mechanisms.METHODS(1)Twenty type 2 diabetic db/db mice were randomly divided into model group and dapagliflozin group,and 10 normal db/m mice were randomly selected as control group.After 40 d of feeding,the mice were killed by anesthesia overdose,the eyeballs were quickly removed for blood collection,the serum was separated,and thoracic aortic tissues were collected.Morphological changes of thoracic aortic tissues were observed by HE staining,and the protein expression levels of ATP synthase alpha subunit(ATP5A1),endothelin-1(ET-1)and prostaglandin I_(2)(PGI_(2))in mouse thoracic aortic tissues were detected by Western blot.(2)HUVECs were divided into control group,DMSO group,high glucose group and high glucose+dapagliflozin group.The fluorescence intensity of reactive oxygen species(ROS)and mitochondrial ATP in HUVECs was detected with ROS and mitochondrial ATP fluorescent probes.The protein expression levels of ATP5A1,ET-1 and PGI_(2)in HUVECs were detected by immunocytochemistry and Western blot.RESULTS Dapagliflozin significantly attenuated thoracic aortic tissue injury in db/db mice.Compared with high glucose group,significantly decreased ROS fluorescence intensity and significantly increased mitochondrial ATP fluorescence intensity in HUVECs were observed in high glucose+dapagliflozin group(both P<0.01).Immunocytochemistry and Western blot results showed that the protein expression levels of ATP5A1 and PGI_(2)were significantly increased,and the protein expression level of ET-1 was significantly decreased after dapagliflozin treatment(all P<0.01).After transfection with an ATP5A1 overexpression plasmid,the protein expression levels of ATP5A1 and PGI_(2)in HUVECs were significantly increased,while the protein expression level of ET-1 was significantly decreased(all P<0.01).CONCLUSION Dapagliflozin protects against thoracic aortic endothelial cell injury in type 2 diabetic mice and injury of HUVECs induced by high glucose,which may be related to the regulation of ATP5A1 expression to improve mitochondrial function.
作者
周小莉
路晰媗
梁海艳
赵丹
雷红
ZHOU Xiaoli;LU Xixuan;LIANG Haiyan;ZHAO Dan;LEI Hong(Department of Endocrinology,Cardiovascular and Cerebrovascular Disease Hospital,Ningxia Medical University General Hospital,Yinchuan 750000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2023年第3期425-433,共9页
Chinese Journal of Pathophysiology
基金
宁夏自然科学基金资助项目(No.2020AAC03372)。
关键词
达格列净
血管内皮损伤
2型糖尿病
ATP合酶α亚基
线粒体
dapagliflozin
vascular endothelial injury
type 2 diabetes mellitus
ATP synthase subunit alpha
mitochondria