中国临床药理学杂志2024，Vol.40Issue(14)：2033-2037,5.DOI:10.13699/j.cnki.1001-6821.2024.14.007
糖尿病小鼠来源的骨髓间充质干细胞培养上清液诱发胰岛素抵抗的研究
Study on insulin resistance induced by supernatant of bone marrow mesenchymal stem cells derived from diabetic mice
摘要
Abstract
Objective To investigate the role of bone marrow mesenchymal stem cells derived from diabetic mice and their paracrine roles in inducing insulin resistance(IR).Methods The mouse model of diabetes mellitus was established,bone marrow mesenchymal stem cells(BMSC)were extracted and cultured,and the culture supernatant(M-BMSC-CS)was collected.(1)Cell experiment:HepG2 hepatocytes were divided into normal low-glycemic culture group[cultured with low-glycemic DMEM(5.55 mmol·L-1)],M-BMSC-CS experimental group(M-BMSC-CS 75 μL),and high-glycemic and high-lipid control group(given 25 mmol·L-1 high-glycemic DMEM+0.25 mmol·L-1 palmitic acid);(2)Animal experiments:Mice were divided into normal mice group(0.9％NaCl by intraperitoneal injection)and M-BMSC-CS-m group(M-BMSC-CS by intraperitoneal injection of normal mice(injection dose 0.2 mL/10 g)].Glucose intake was measured by glucose oxidase method.The fluorescence intensity of Glut2 protein was detected by immunofluorescence.The expression of insulin signaling pathway protein was detected by Western blot.Test oral glucose tolerance(OGTT)and insulin tolerance(ITT).Results The glucose intakes of the normal low-glucose culture group,the M-BMSC-CS experimental group and the high-glucose and high-lipid control group were(2.96±0.05),(1.64±0.28)and(1.42±0.32)mmol·L-1,respectively;the fluorescence expressions of glucose transporter 2(Glut2)were 53.21±2.70,30.95±3.39 and 34.96±7.60,respectively;the protein expression levels of phosphorylated insulin receptor substrate 1-ser307(p-IRS-1ser307)were 0.46±0.21,1.09±0.24 and 0.91±0.16,respectively;phosphorylated protein kinase(p-AKT)protein expression levels were 0.94±0.05,0.59±0.06 and 0.53±0.05;Glut2 protein expression levels were 1.08±0.14,0.58±0.14 and 0.62±0.09,respectively.The above indexes in M-BMSC-CS experimental group were statistically significant compared with those in normal low-glycemic culture group(all P＜0.05).Fasting blood glucose levels in the normal group and M-BMSC-CS-m group were(5.23±0.57)and(9.30±1.14)mmol·L-1;p-AKT protein expression level were 1.27±0.21 and 0.51±0.19;Glut2 protein expression level were 1.17±0.17 and 0.79±0.09,respectively.The above indexes in M-BMSC-CS-m group were significantly different from those in normal mouse group(P＜0.05).Conclusion BMSC culture supernatant from diabetic mice induced insulin resistance of normal HepG2 hepatocytes in vitro and normal mice in vivo.关键词
糖尿病小鼠/骨髓间充质干细胞培养上清液/胰岛素抵抗/HepG2肝细胞/正常小鼠Key words
diabetic mice/bone marrow mesenchymal stem cell culture supernatant/insulin resistance/HepG2 liver cell/normal mice分类
医药卫生引用本文复制引用
李包娟,周克春,阿布都拉·米热合买提,祖丽胡马·热合曼,叶雨萌,张䶮之..糖尿病小鼠来源的骨髓间充质干细胞培养上清液诱发胰岛素抵抗的研究[J].中国临床药理学杂志,2024,40(14):2033-2037,5.基金项目
国家自然科学基金资助项目(81760767) （81760767）
新疆天然药物活性组分与释药技术重点实验室基金资助项目(XJDX1713) （XJDX1713）
