Caloric Restriction Regulates Mouse Dendritic Cell Migration Through The Lok-ezrin Pathway

Objective: To investigate the regulation of caloric restriction on the migration ability of mouse dendritic cell and the mechanism。Methods:(1)Comparing the volume of melanin in C57 BL/6 mice after ad libitum diet and caloric restriction.(2)Differentially expressed genes in spleen dendritic cells(DC)of mice after calorie restriction were screened by third-generation sequencing;(3)Proliferation of DC2.4 cells in control group and caloric restriction group was detected by CCK-8asssay and flow cytometry;(4)The expression of Lok mRNA in DC2.4cells of control group and caloric restriction group was detected by q PCR;(5)The expression of Lok and Ezrin in DC2.4 cells of control group and calorie restriction group was detected by Western Blot assay;(6)The effects of Lok inhibitor SB633825 on Ezrin expression in DC2.4 cells was detected by immunofluorescence assay(IF),and the effects of overexpression of Lok on Ezrin expression in DC2.4 cells was detected by IF;(7)The effect of caloric restriction and SB633825 on DC2.4 cell migration was detected by scratch test.Results:(1)The volume of melanin tumor decreased after caloric restriction;(2)Sequencing results showed that Lok gene was up-regulated in mouse DC cells after caloric restriction;(3)Caloric restriction promoted proliferation of DC2.4 cells;(4)The expression of Lok mRNA in DC2.4 cells increased after caloric restriction;(5)The expression of Lok and Ezrin in DC2.4 cells increased after calorie restriction;(6)Inhibition of calorict restriction by SB633825 up-regulated Ezrin expression in DC2.4 cells,and overexpression of Lok up-regulated Ezrin expression in DC2.4 cells.(7)Caloric restriction inhibited the migration of DC2.4 cells and SB-633825 can reverse this inhibition.Conclusions:(1)Caloric restriction has antitumor effects.(2)Caloric restriction can promote the immune function of mouse dendritic cells by upregulating the expression of Lok in DC cells,promoting Ezrin expression and changing its distribution in DC cells,and regulating DC migration.