丁然 1王欣笛 1吴磊 2高林荫 1史宝宝 2王海涛3
作者信息
- 1. 华北理工大学,基础医学院,河北省慢性疾病基础医学重点实验室,唐山市慢性病临床基础研究重点实验室,唐山 063200
- 2. 华北理工大学,心理与精神卫生学院,河北省心理健康与脑科学重点实验室,唐山 063200
- 3. 华北理工大学,基础医学院,河北省慢性疾病基础医学重点实验室,唐山市慢性病临床基础研究重点实验室,唐山 063200||华北理工大学,心理与精神卫生学院,河北省心理健康与脑科学重点实验室,唐山 063200
- 折叠
摘要
Abstract
Objective:To investigates the role of Wnt/β-catenin signaling pathway in the phenotypic transformation of astrocytes in diabetic encephalopathy.Methods:A diabetic mouse model was established by feeding the mice with high-fat diet for 12 weeks.Twelve C57BL/6 mice were randomly divided into control group and diabetic groups.The blood glucose level and body weight of the mice were detected at 0 and 12 weeks.Immunofluorescence was used to detect the expression of A1 astrocyte marker C3 and A2 astrocyte marker S100A10 in the hippocampus.C8-D1A astrocyte cell line was used to establish three groups:control,palmitic acid(PA)-treated,and PA+Wnt pathway activator(SKL2001).cell viability at various PA concentrations(0,50,100,200,300 μmol/L)was assessed through the CCK-8 assay.Lipid accumulation in C8-D1A astrocytes was evaluated by using Oil Red O staining.Western blotting analysis was performed to measure the protein expression levels of GFAP,C3,S100A10,Wnt3a,β-catenin,p-GSK3β and GSK3β.Immunofluorescence was used to visualize the expression of C3/GFAP,S100A10/GFAP,Wnt3a and β-catenin in astrocytes.Results:Compared with the control group,the blood glucose level and body weight of the diabetic group were significantly increased.The fluorescence intensity of C3 in the hippocampus of diabetic mice was significantly higher than that of the control group,but the fluorescence intensity of S100A10 was not significantly different between the two groups.Compared with the control group,PA treatment significantly inhibited the cell viability in a concentration-dependent manner.Oil Red O staining showed orange-red lipid droplets in the cytoplasm of C8-D1A cells in PA-treated groups.Western blotting results showed that the protein expressions of GFAP and C3 in the PA group were significantly higher than those in the control group,while the protein expressions of Wnt3a,β-catenin,and p-GSK3β/GSK3β in the PA group were significantly lower than those in the control group.In the PA+SKL2001 group,SKL2001 partially reversed the PA-induced changes in protein expression.No significant differences were found in S100A10 protein expression among the three groups.Immunofluorescence results confirmed that compared with the control group,the fluorescence intensity of C3/GFAP in the PA group was increased.Compared with the PA group,the fluorescence intensity of C3/GFAP was decreased in the PA+SKL2001 group,while the fluorescence intensity of S100A10 did not differ significantly between groups.Compared with the control group,the fluorescence intensity of Wnt3a and the translocation of β-catenin into the nucleus in the PA group decreased.Compared with the PA group,both indicators above were increased in the PA+SKL2001 group.Conclusion:In diabetic encephalopathy,the transformation of astrocyte phenotype may be caused by inhibiting the expression of Wnt/β-catenin signaling pathway-related proteins,leading to the transformation of astrocytes into type A1,thereby exacerbating disease progression.关键词
糖尿病脑病/星形胶质细胞/棕榈酸/Wnt/β-catenin信号通路Key words
diabetic encephalopathy/astrocyte/palmitic acid/Wnt/β-catenin signaling pathway分类
医药卫生
