| 阿勒西尔·努尔买买提,王 婷,努尔买买提江·买合木提,苏丽亚·迪力夏提,郑 敏.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[J].现代生物医学进展英文版,2024,(17):3217-3225. |
| 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 |
| Received:March 03, 2024 Revised:March 26, 2024 |
| DOI：10.13241/j.cnki.pmb.2024.17.004 |
| 中文关键词: 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 |
| 英文关键词: Propofol Psychological dependence Nervous immune system C-C chemokine receptor 5-IP3 signaling pathways Toll-like receptor 4 -nuclear factor κB-P38 signaling pathways |
| 基金项目:新疆维吾尔自治区自然科学基金项目（2021D01C390） |
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| 中文摘要: |
| 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15 μM干预组CCL2、CCL5和TNF-α水平升高（P<0.05）；丙泊酚20 μM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比较差异具有统计学意义（P<0.05）；与丙泊酚干预组比较，IBU预处理+丙泊酚干预组也表现出降低移动距离趋势（P<0.05）。在大鼠前叶额皮质组织中，与空白组比较，丙泊酚干预组CCL2、 CCL5和TNF-α水平显著增加（P<0.05）。与丙泊酚干预组比较，IBU预处理+丙泊酚干预组CCL2、CCL5和TNF-α水平显著降低（P<0.05）。与空白组比较，丙泊酚干预组前额叶皮质组织中TLR4蛋白表达、p-p65、p-ERK1/2显著升高（P<0.05）。与丙泊酚干预组比较，IBU预处理+丙泊酚干预组的前额叶皮质组织中TLR4蛋白表达、p-p65、p-ERK1/2显著降低（P<0.05）。结论：TLR4-NFκB-P38及CCR5-IP3信号通路可能参与神经免疫调节在丙泊酚药物依赖中的作用。 |
| 英文摘要: |
| ABSTRACT Objective: 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-α in 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-ERK1/2 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-ERK1/2 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. |
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