维生素D通过哺乳动物STE20样激酶1-核因子E2相关因子2通路调控自噬抑制缺氧/复氧诱导的糖尿病心肌细胞损伤的研究  

Vitamin D inhibits hypoxia/reoxygenation⁃induced cardiomyocytes injury in hyperglycemic conditions by regulating autophagy through Mst1⁃Nrf2 pathway

作　　者：厚荣荣 孔颖[1] 高雷[1] 陶美花 徐静[1] 尹涛 HOU Rongrong;KONG Ying;GAO Lei(Department of Endocrinology,The Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China)

机构地区：[1]西安交通大学第二附属医院内分泌科,710004 [2]空军军医大学第一附属医院心血管内科

出　　处：《中国糖尿病杂志》2023年第10期777-782,共6页Chinese Journal of Diabetes

基　　金：国家自然科学基金(82000786);陕西省重点研发计划项目(2020SF-248)。

摘　　要：目的探讨维生素D(VitD)通过哺乳动物STE20样激酶1(Mst1)-核因子E2相关因子2(Nrf2)通路调控自噬抑制缺氧/复氧(H/R)诱导的DM心肌细胞损伤。方法大鼠H9C2细胞分为正常对照组(Con)、葡萄糖浓度25.5 mmol/L(高糖)组(HG)、H/R组、HG+H/R组、HG+H/R+VitD组、HG+H/R+3-MA组、HG+H/R+Mst1抑制剂组(HG+H/R+Mst1i组)。CCK8检测细胞活性,TUNEL检测细胞凋亡率,Western blot法检测选择性自噬接头蛋白(p62)、重组人自噬效应蛋白1(Beclin1)、Mst1、p-Mst1、Nrf2的蛋白表达。结果与Con组比较,H/R、HG、HG+H/R组凋亡率、切割的半胱氨酸蛋白酶3(c-Caspase-3)蛋白表达升高(P<0.05),细胞活力降低(P<0.05)。与HG+H/R组比较,HG+H/R+VitD组细胞活力升高(P<0.05),凋亡率、c-Caspase-3蛋白表达降低(P<0.05)。与Con组比较,HG+H/R组Beclin1、Mst1、p-Mst1蛋白表达升高(P<0.05),Nrf2、p62蛋白表达降低(P<0.05)。与HG+H/R组比较,HG+H/R+3-MA组p62蛋白表达升高(P<0.05),Beclin1蛋白表达降低(P<0.05),HG+H/R+VitD、HG+H/R+Mst1i组Nrf2、p62蛋白表达升高(P<0.05),Mst1、p-Mst1、Beclin-1蛋白表达降低(P<0.05)。结论VitD通过调控自噬减少HG+H/R诱导的心肌细胞凋亡,可能与Mst1-Nrf2通路相关。Objective To investigate whether VitD can inhibit H/R induced cardiomyocytes injury in hyperglycemic conditions by regulating autophagy through Mst1-Nrf2.Methods Rat H9C2 cells were divided into the following groups:control group(Con),high glucose group(HG),H/R group,HG+H/R group,HG+H/R+VitD group,HG+H/R+3-MA group and HG+H/R+Mst1i group.CCK8 assay was used to detect cell viability.TUNEL assay was used to detect cell apoptosis.The expression of p62,Beclin1,Mst1,p-Mst1 and Nrf2 was detected by Western blot.Results Compared with Con group,apoptotic rate and c-Caspase-3 expression in H/R,HG and HG+H/R groups were increased(P<0.05),while cell viability was decreased(P<0.05).Compared with HG+H/R group,cell viability in HG+H/R+VitD group was increased(P<0.05),while apoptotic rate and c-Caspase-3 expression were decreased(P<0.05).Compared with Con group,in HG+H/R group,the protein expression of Beclin1,Mst1 and p-Mst1 was increased(P<0.05)and the protein expression of Nrf2 and p62 was decreased.Compared with HG+H/R group,the protein expression of p62 was increased while Beclin-1 was decreased in HG+H/R+3-MA group(P<0.05).Compared with HG+H/R group,the protein expression of Nrf2 and p62 was increased while Mst1,p-Mst1 and Beclin-1 was decreased in HG+H/R+VitD and HG+H/R+Mst1i groups(P<0.05).Conclusion VitD reduces HG+H/R-induced apoptosis of cardiomyocytes by regulating autophagy,which may be related to the Mst1-Nrf2 pathway.

关 键 词：维生素D 糖尿病 缺氧/复氧 心肌细胞 凋亡 

分 类 号：R587.2[医药卫生—内分泌]