TLR4-NFκB-P38及CCR5-IP3信号通路对丙泊酚药物依赖大鼠胶质细胞神经免疫的影响  

Effects of TLR4-NFκB-P38 and CCR5-IP3 Signaling Pathways on Neuroimmunity of Glial Cells in Propofol-Dependent Rats

作　　者：阿勒西尔·努尔买买提 王婷[1] 努尔买买提江·买合木提[1] 苏丽亚·迪力夏提 郑敏 Alexir·Nuermaimaiti;WANG Ting;Nuermaimaitijiang·Maihemuti;Sulia·Dilixiati;ZHENG Min(Anesthesia and Perioperative Medical Center,Cancer Hospital Affiliated to Xinjiang Medical University,Urumqi,Xinjiang,830011,China)

机构地区：[1]新疆医科大学附属肿瘤医院麻醉与围术期医学中心,新疆乌鲁木齐830011

出　　处：《现代生物医学进展》2024年第17期3217-3225,共9页Progress in Modern Biomedicine

基　　金：新疆维吾尔自治区自然科学基金项目(2021D01C390)。

摘　　要：目的:探讨Toll样受体4(TLR4)-核因子κB(NFκB)-P38及C-C趋化因子受体5(CCR5)-(三磷酸肌醇)IP3信号通路对丙泊酚药物依赖大鼠胶质细胞神经免疫的影响。方法:体外实验:使用细胞计数试剂-8(CCK-8)试剂盒检测并比较丙泊酚药物依赖对BV2(小鼠小胶质细胞)细胞活性的影响;流式细胞术比较丙泊酚药物依赖对BV2细胞凋亡率的影响;比较不同浓度丙泊酚对BV2细胞因子[肿瘤坏死因子-α(TNF-α)、趋化因子配体2(CCL2)和CCL5]的影响。采用免疫蛋白印记法(WB)检测并比较TLR4、p65、磷酸化-p65(p-p65)、细胞外调节蛋白激酶(ERK)1/2和p-ERK1/2蛋白表达。动物实验:利用24只SD大鼠建立丙泊酚药物依赖模型分为三组:空白组(注射等量生理盐水,n=8),丙泊酚药物依赖模型组(注射40 mg/kg丙泊酚+生理盐水,n=8),异丁司特(IBU)预处理+丙泊酚干预组(注射7.5 mg/kg IBU,n=8)。采用条件性位置偏爱(CPP)和移动距离评估丙泊酚所致的成瘾效应。检测并比较SD大鼠模型血清细胞因子(CCL2、CCL5和TNF-α)水平以及前额叶皮质组织中相关蛋白表达。结果:与空白组比较,丙泊酚15μM干预组CCL2、CCL5和TNF-α水平升高(P<0.05);丙泊酚20μM干预组CCL2、CCL5和TNF-α水平升高(P<0.05)。与空白组比较,丙泊酚干预组CCL2、CCL5和TNF-α水平升高(P<0.05),与丙泊酚干预组对比,IBU预处理+丙泊酚干预组的CCL2、CCL5和TNF-α水平显著下降(P<0.05)。与空白组比较,丙泊酚干预组和IBU预处理+丙泊酚干预组的TLR4蛋白表达显著增加,p-p65和p-ERK1/2蛋白表达显著上调(P<0.05),而p65和ERK1/2的蛋白表达变化差异无统计学意义(P>0.05)。与丙泊酚干预组比较,IBU预处理+丙泊酚干预组TLR4、p-p65和p-ERK1/2蛋白表达显著降低(P<0.05)。与空白组比较,丙泊酚干预组CPP值显著升高(P<0.05),与丙泊酚干预组比较,IBU预处理+丙泊酚干预组CPP值显著降低(P<0.05)。与空白组比较,丙泊酚干预组的移动距离组间�Objective:To investigate the effects of Toll-like receptor 4(TLR4)-nuclear factorκB(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-αin propofol 15μM intervention group were increased(P<0.05).The levels of CCL2,CCL5 and TNF-αin propofol 20μM intervention group were increased(P<0.05).Compared with blank group,the levels of CCL2,CCL5 and TNF-αin 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+pr

关 键 词：丙泊酚 精神依赖 神经免疫系统 CCR5-IP3信号通路 TLR4-NFκB-P38信号通路 

分 类 号：R-33[医药卫生] R965