EPDR1对肝细胞脂质沉积的影响

Effect of EPDR1 on hepatocyte lipid deposition

目的:检测室管膜素相关蛋白1(EPDR1)在野生型C57BL/6小鼠各组织中的表达情况;观察2型糖尿病状态下,小鼠各组织中EPDR1的表达;探讨EPDR1对AML12肝细胞脂质沉积的影响。方法:采用Western blot检测C57BL/6小鼠心、肝、脾、肺、肾、腓肠肌、棕色脂肪以及脑组织中EPDR1蛋白的表达;Western blot和免疫组织化学(IHC)染色比较db/db小鼠与C57BL/6小鼠肝、腓肠肌、心和肾组织中EPDR1蛋白表达情况。采用油酸和棕榈酸联合诱导AML12细胞建立脂质沉积模型,且经腺病毒过表达EPDR1和给予外源重组蛋白EPDR1(rEPDR1)干预,ELISA测定胞内甘油三酯(TG)含量,油红O染色评估EPDR1对AML12细胞脂质沉积的影响。结果:Western blot和IHC均显示EPDR1广泛表达于野生型小鼠心、肝、脾、肺、肾、腓肠肌、棕色脂肪和脑组织中,且在肝脏中蛋白表达最高;而在糖尿病db/db小鼠肝、腓肠肌、心和肾组织中较野生型小鼠表达下调;油红O染色结果显示,过表达EPDR1或给予rEPDR1均可减少AML12肝细胞中脂质沉积,且TG含量较模型组下降(P<0.05)。结论:EPDR1广泛表达在野生型小鼠各个组织,且在糖尿病小鼠肝脏组织中表达下调,而上调EPDR1可以缓解肝细胞中的脂质沉积。本研究为进一步揭示EPDR1在糖尿病肝组织脂肪变中的作用奠定实验基础。

AIM:This study aims to examine the ependymin-related protein 1(EPDR1)expression in various tissues from wild-type C57BL/6 mice and type 2 diabetes(db/db)mice.The impact of EPDR1 on lipid accumulation in alpha mouse liver 12(AML12)hepatocytes was also investigated.METHODS:Western blot was used to detect EPDR1 protein expression in the heart,liver,spleen,lung,kidney,gastrocnemius,brown adipose and brain tissues of C57BL/6 mice.Western blot and immunohistochemical(IHC)staining were also used to compare EPDR1 protein expression in the liver,gastrocnemius muscle,heart and kidney tissues of db/db and C57BL/6 mice.To develop an AML12 cell lipid deposition model,palmitic acid(PA)+oleic acid(OA)was used,and the cells were transfected with adenovirus overexpressing EPDR1 or treated with exogenous recombinant EPDR1 protein(rEPDR1).ELISA was conducted to determine intracellular triglyceride(TG)content,and oil red O staining was employed to assess the effect of EPDR1 on lipid accumulation in AML12 cells.RESULTS:Western blot and IHC staining results revealed that EPDR1 was widely expressed in various tissues of wild-type mice,with the liver exhibiting the highest protein expression level.However,EPDR1 expression was down-regulated in the liver,gastrocnemius muscle,heart and kidney tissues in diabetic db/db mice compared with wild�type mice.Oil red O staining revealed that overexpression of EPDR1 in AML12 liver cells or rEPDR1 treatment led to reduced lipid accumulation.Furthermore,the TG content significantly decreased compared with the model group(P<0.05).CONCLUSION:EPDR1 is expressed in various tissues of wild-type mice,but showed diminished expression in the liver tissues of diabetic mice.Nevertheless,enhancing the expression of EPDR1 can aid in reducing lipid accumulation in hepatocytes.These findings provide an experimental foundation for further exploration of the role of EPDR1 in the development of fatty liver in diabetic liver tissue.