Effects of low expression of α-glucosidase on morphology,proliferation,inflammatory response and oxidative stress response of pancreatic β cells
Objective To observe the effects of low expression of α-glucosidase(GAA)on morphology,prolifera-tion,inflammatory response,and oxidative stress response of pancreatic β-cells in order to investigate the potential role of GAA in the pathogenesis of diabetes mellitus.Methods The expression of GAA in βTC6 cells was interfered by siR-NA1,2,3 and 4,respectively.As a result,the interference effect of siRNA4 was the best,and it was used as the interfer-ence sequence in subsequent experiments.βTC6 cells were randomly divided into the siRNA group,negative control group(NC group)and blank control group(Con group),respectively.Cells in the siRNA group were transfected with siR-NA4 sequence to reduce the expression of GAA,cells in the negative control group were transfected with siRNANC se-quence,and cells in the Con group were not treated.After 24,48,72 and 96 h of cell culture,the cell proliferation of each group was detected by CCK-8.They were cultured for 48 h,the number and morphology of cells in each group were observed by inverted microscope,the inflammatory factors IL-6 and IL-1β protein in cells were detected by Western blot-ting,and the content of reactive oxygen species(ROS)in cells was detected by immunofluorescence.Results Com-pared with the Con and NC groups,the cells increased,the intercellular space became smaller under the microscope,the OD values of cells increased at 24,48,72,and 96 h of culture,and the levels of IL-6 and ROS decreased after 48 h of cul-ture in the siRNA group(all P<0.01).No significant differences were found in these indicators between the Con and NC groups(all P>0.05).Conclusions Low expression of GAA can lead to alterations in cell morphology,enhanced prolif-erative capacity,and diminished inflammatory and oxidative stress responses of pancreatic β-cells.GAA may participate in the pathogenesis of diabetes by changing the morphology of pancreatic β-cells and enhancing inflammatory and oxidative stress responses.
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