丹皮酚上调miR⁃223⁃3p减轻高糖诱导的小鼠心肌微血管内皮细胞损伤的研究  

Upregulation of paeonol on miR⁃223⁃3p alleviates endothelial cell damage induced by high glucose

作　　者：邓波[1] 舒远 胡玲[1] DENG Bo;SHU Yuan;HU Ling(Department of Endocrinology,The Third Affiliated Hospital of Nanchang University,Nanchang 330008,China)

机构地区：[1]南昌大学第三附属医院内分泌科,330008 [2]南昌大学医学部第二临床医学院

出　　处：《中国糖尿病杂志》2023年第9期697-702,共6页Chinese Journal of Diabetes

基　　金：江西省中医药管理局科技计划(2021A372);江西省重点研发计划项目(20203BBG73053);江西省卫生计生委科技计划(202311215)。

摘　　要：目的探讨丹皮酚(Pae)对高糖诱导的小鼠心肌微血管内皮细胞(MCMECs)的影响及其作用机制。方法建立HG诱导MCMECs体外模型,分为正常对照组(Con)组、高糖(HG)组、HG+Pae30μmol/L组(HG+Pae30)、HG+Pae60μmol/L组(HG+Pae60)、HG+Pae120μmol/L组(HG+Pae120)。将miR⁃NC、miR⁃223⁃3p mimics、anti⁃miR⁃NC、anti⁃miR⁃223⁃3p转染细胞,分为HG+miR⁃NC、HG+miR⁃223⁃3p组及HG+Pae60+anti(-)、HG+Pae60+anti(+)组。MTT法检测细胞活力,流式细胞仪检测各组细胞凋亡率,RT⁃PCR检测miR⁃223⁃3p、NLRP3 mRNA表达。Western blot法检测Nod样受体热蛋白结构域相关蛋白3(NLRP3)、B淋巴细胞瘤⁃2(Bcl⁃2)、Bcl⁃2相关X蛋白(Bax)、胱天蛋白酶⁃3(Caspase⁃3)蛋白表达。结果60μmol/L Pae改善高糖诱导细胞损伤,用于后续实验。与Con组比较,HG组NLRP3 mRNA、NLRP3、Bax、Caspase⁃3蛋白升高(P<0.05),miR⁃223⁃3p、Bcl⁃2蛋白表达降低(P<0.05)。与HG组比较,HG+Pae60组miR⁃223⁃3p、Bcl⁃2蛋白表达升高(P<0.05),NLRP3 mRNA、NLRP3、Bax、Caspase⁃3蛋白表达降低(P<0.05)。与HG+miR⁃NC组比较,HG+miR⁃223⁃3p组细胞活力、Bcl⁃2蛋白表达升高(P<0.05),细胞凋亡率、NLRP3、Bax、Caspase⁃3蛋白表达降低(P<0.05);与HG+Pae60+anti(-)组比较,HG+Pae60+anti(+)组细胞凋亡率、NLRP3、Bax、Caspase⁃3蛋白表达升高(P<0.05),细胞活力、Bcl⁃2蛋白表达降低。结论Pae可能通过上调miR⁃223⁃3p减轻高糖诱导内皮细胞损伤,并抑制其凋亡。Objective To investigate the effects of Paeonol(Pae)on myocardial microvascular endothelial cells(MCMECs)induced by high glucose(HG)and its mechanism.Methods An in vitro model of HG⁃induced MCMECs was established,and cells were divided into normal control(Con),HG group,HG+Pae30μmol/L(HG+Pae30),HG+Pae60μmol/L(HG+Pae60),and HG+Pae120μmol/L(HG+Pae120)group.Cells were transfected with miR⁃NC,miR⁃223⁃3p mimics,anti⁃miR⁃NC,anti⁃miR⁃223⁃3p,and divided into HG+miR⁃NC group,HG+miR⁃223⁃3p group,HG+Pae60+anti(-)group and HG+Pae60+anti(+)group.MTT assay was performed to detect cell viability.Flow cytometry was used in detection of cell apoptosis rate in each group.RT⁃PCR was performed to detect the expression of miR⁃223⁃3p and NLRP3 mRNA.Western blot was performed to detect the protein levels of NLRP3 and B lymphoblastoma⁃2(Bcl⁃2),Bcl⁃2 associated X protein(Bax),Caspase⁃3.Results Paeimproved HG⁃induced cell damage with a concentration of 60μmol/L,and was used in subsequent experiments.Compared with Con group,expression of NLRP3,Bax and Caspase⁃3 increased,while expression of miR⁃223⁃3p and Bcl⁃2 decreased in HG group(P<0.05).Compared with HG group,miR⁃223⁃3p and Bcl⁃2 expression increased,while NLRP3,Bax,Caspase⁃3 protein expression decreased in HG+pae60 group(P<0.05).Compared with HG+miR⁃NC group,cell activity and Bcl⁃2 expression increased,while apoptosis rate,NLRP3,Bax,and Caspase⁃3 protein expression decreased in HG+miR⁃223⁃3p group(P<0.05).Compared with HG+Pae60+anti(-)group,apoptosis rate,NLRP3,Bax,Caspase⁃3 protein expression increased,while cell activity Bcl⁃2 expression decreased in HG+Pae60+anti(+)group(P<0.05).Conclusion Pae may reduce HG⁃induced endothelial cell damage and inhibit apoptosis by up⁃regulating miR⁃223⁃3p.

关 键 词：丹皮酚 高糖 miR⁃223⁃3p Nod样受体热蛋白结构域相关蛋白3 内皮细胞 

分 类 号：R285.5[医药卫生—中药学]