Effects of TLR4-NFκB-P38 and CCR5-IP3 Signaling Pathways on Neuroimmunity of Glial Cells in Propofol-Dependent Rats
Objectives:To investigate the effects of Toll-like receptor 4(TLR4)-nuclear factor κB(NFκB)-P38 and C-C chemokine receptor 5(CCR5)-(inositol triphosphate)IP3 signaling pathways on the neuroimmunity of glial cells in propofol-dependent rats.Methods:In vitro experiments:the effect of propofol drug dependence on BV2(mouse microglia)cell activity were detected and compared by cell counting kit-8(CCK-8)kit.The effect of propofol drug dependence on the apoptosis rate of BV2 cells were compared by flow cytometry.The effects of different concentrations of propofol on BV2 cytokines[tumor necrosis factor-α(TNF-α),chemokine ligand 2(CCL2)and CCL5]were compared.The expressions of TLR4,p65,phosphorylated-p65(p-p65),extracellular regulated protein kinase(ERK)1/2 and p-ERK1/2 were detected and compared by Western blot(WB).Animal experiments:24 SD rats were used to establish a propofol drug dependence model and divided into three groups:blank group(injected with the same amount of normal saline,n=8),propofol drug dependence model group(injected with 40 mg/kg propofol and normal saline,n=8),isobuprost(IBU)pretreatment+propofol intervention group(injected with 7.5 mg/kg IBU,n=8).The addictive effect of propofol were evaluated by conditioned place preference(CPP)and moving distance.The levels of serum cytokines(CCL2,CCL5 and TNF-α)and the expression of related proteins in prefrontal cortex of SD rat model were detected and compared.Results:Compared with blank group,the levels of CCL2,CCL5 and TNF-α in propofol 15 μM intervention group were increased(P＜0.05).The levels of CCL2,CCL5 and TNF-α in propofol 20 μM intervention group were increased(P＜0.05).Compared with blank group,the levels of CCL2,CCL5 and TNF-α in propofol intervention group were increased(P＜0.05),compared with propofol intervention group,the levels of CCL2,CCL5 and TNF-αin IBU pretreatment+propofol intervention group were significantly decreased(P＜0.05).Compared with blank group,the expression of TLR4 protein in propofol intervention group and IBU pretreatment+propofol intervention group was significantly increased,and the expression of p-p65 and p-ERKl/2 protein was significantly up-regulated(P＜0.05),there was no significant difference in the expression of p65 and ERK1/2 protein(P＞0.05).Compared with propofol intervention group,the expression of TLR4,p-p65 and p-ERKl/2 protein in IBU pretreatment+propofol intervention group was significantly decreased(P＜0.05).Compared with blank group,the CPP value in propofol intervention group was significantly increased(P＜0.05),compared with propofol intervention group,the CPP value in IBU pretreatment+propofol intervention group was significantly decreased(P＜0.05).Compared with blank group,the difference in the moving distance between propofol intervention group and blank group was statistically significant(P＜0.05).Compared with propofol intervention group,the IBU pretreatment+propofol intervention group also showed a tendency to reduce the moving distance(P＜0.05).In the frontal cortex of rats,compared with blank group,the levels of CCL2,CCL5 and TNF-α in propofol intervention group were significantly increased(P＜0.05).Compared with propofol intervention group,the levels of CCL2,CCL5 and TNF-α in IBU pretreatment+propofol intervention group were significantly decreased(P＜0.05).Compared with blank group,the expression of TLR4 protein,p-p65 and p-ERK1/2 in prefrontal cortex of the propofol intervention group were significantly increased(P＜0.05).Compared with propofol intervention group,the expression of TLR4 protein,p-p65 and p-ERK1/2 in prefrontal cortex of the IBU pretreatment+propofol intervention group were significantly decreased(P＜0.05).Conclusion:TLR4-NFκB-P38 and CCR5-IP3 signaling pathways may be involved in the role of neuroimmune regulation in propofol drug dependence.
NETL
NSTL
万方数据
