Correlation Study Between Autophagy And Apoptosis Of Peritoneal Macrophages And The Changes Of Immune Function In Sepsis Mice

Objective:To explore the relationship between autophagy,apoptosis and immune function changes of sepsis in mice peritoneal macrophages,in order to reveal the mechanism of the occurrence and development of sepsis and provide a theoretical basis,lay the theoretical foundation for the exploration of the new scheme for the treatment of sepsis.Methods:A total of 72 male Balb/c mice were fasted for 12 hours before experiment,free drinking water,were randomly divided into 6 groups,12 rats in each group,and divided into LPS+2h group,LPS+6h group,LPS+12h group,LPS+24h group and LPS+36h group.the experimental group of mice abdominal local routine disinfection of skin after intraperitoneal injection of LPS,dosage of 20mg/kg.(every mice injected with LPS solution volume is about 200-250ul,The normal control group were injected with the same dose of saline..ELISA method was used to detect the concentrations of inflammatory factors IL-2,IL-10 and TNF-in peripheral blood,and the CD4~+T/CD~+8 T ratio in peripheral blood was detected by flow cytometry;Detection of LC3II?Beclin-1/beta-action expression in macrophages of mice by Western Blot and the level of autophagy.Results:(1)The expression of LC3II and Beclin-1 in peritoneal macrophages of mice in group LPS+2h was significantly higher than that in normal group(p<0.05),subsequently,as time went on,the expression of LC3II and Beclin-1 in peritoneal macrophages of group LPS+6h,group LPS+12h,group LPS+24h and group LPS+36h showed a trend of continuous decrease compared with normal control mice(P<0.05)(2)LPS+2h,LPS+6h group,the peripheral blood CD4~+T/CD8~+T cell ratio was significantly higher than the normal group(p<0.05),6h peak,and Then,as time went on,the ratio of CD4~+T/CD8~+T cells in peripheral blood of group LPS+12h,group LPS+24h and group LPS+36h showed a trend of continuous decrease compared with normal control mice(P<0.05)(3)The concentrations of IL-2 and Tnf-?in peripheral blood serum of LPS+2h,LPS+6h,LPS+12h group were significantly higher than those in normal group(p<0.05)12h reached the peak,As time goes on,The concentrations of IL-2 and TNF-a in peripheral blood of group LPS+24h and group LPS+36h showed a trend of continuous decrease compared with normal control mice(p<0.05),LPS+2h,6h,12h,24h,36h peripheral serum IL-10 concentration was significantly higher than that of the normal group(p<0.05).(4)LPS+6h,12h,24h,36h group peritoneal macrophages LC3II,Beclin-1and peripheral serum CD~+4T/CD~+8T ratio correlation index R~2=0.716(P=0.043)R~2=0.854(P=0.023).(5)The apoptosis level of LPS+2h group of peritoneal macrophages was slightly increased compared with normal group(P<0.05),The apoptotic rate of peritoneal macrophages in group LPS+6h,group LPS+12h,group LPS+24h and group LPS+36h was significantly higher than that of normal control mice,and showed an upward trend(p<0.05),The correlation between the apoptosis of peritoneal macrophages and the ratio of CD~+4T/CD~+8T in peripheral blood of mice in group LPS+6h,group LPS+12h,group LPS+24h and group LPS+36h was R~2=0.796(P=0.035).Conclusions:(1)The autophagy of peritoneal macrophages in sepsis mice is strongly dependent on the immune function,It has protective effect on the immunity of mice with sepsis.(2)The apoptosis of peritoneal macrophages in sepsis mice has a strong correlation with the immune function,and has a negative regulatory effect on the immune function of sepsis mice.(3)The autophagy of peritoneal macrophages in sepsis mice can regulate the apoptosis of peritoneal macrophages,may be the conversion of autophagy to apoptosiswhich is related to the changes of immune function in mice.