大麻素受体激动剂花生四烯酰氯乙胺(ACEA)通过抑制炎症反应改善小鼠脓毒症相关性脑病

Cannabinoid receptor 1 agonist arachidonyl-2′-chloroethylamide (ACEA) improves sepsis-associated encephalopathy by inhibiting inflammatory factors

目的 探讨大麻素受体激动剂花生四烯酰氯乙胺(ACEA)对脓毒症相关性脑病(SAE)小鼠认知功能的影响。方法C57BL/6小鼠随机分成人工脑脊液(ACSF)组和脂多糖(LPS)组,通过采用LPS侧脑室注射构建SAE模型,小鼠脓毒症严重程度评分(MSS)和体质量变化判断小鼠的基本情况;行为学范式评估小鼠运动能力(旷场实验)、认知功能(情景前置性条件恐惧实验、 Y迷宫实验)。评估ACEA干预效果中,小鼠随机分为ACSF组、 ACEA干预的ACSF组、 LPS组和ACEA干预的LPS组,ACEA干预组给与1.5 mg/kg ACEA,实时定量PCR检测小鼠海马组织白细胞介素1β(IL-1β)、 IL-6、肿瘤坏死因子α(TNF-α)的mRNA表达水平;Western blot法检测小鼠海马组织IL-6、 TNF-α的蛋白表达水平;尼氏染色检测小鼠海马CA1区神经元损伤情况;行为学范式评估小鼠运动能力(旷场实验)、认知功能(情景前置性条件恐惧实验、 Y迷宫实验)。结果 侧脑室注射LPS 3 d后,小鼠存在显著认知功能障碍,证实SAE造模成功。与ACSF组相比,LPS组海马炎症因子IL-6、 TNF-α及IL-1β mRNA表达水平显著升高,IL-6、 TNF-α蛋白表达水平显著增加,CA1区可见神经元尼氏体减少,小鼠存在显著认知功能障碍;与LPS组相比,ACEA干预组IL-6、 TNF-α及IL-1β mRNA表达水平显著降低,IL-6、 TNF-α蛋白表达水平显著减少,神经元尼氏小体增多,小鼠认知功能改善。结论 ACEA通过抑制IL-6、 TNF-α炎症因子表达水平改善SAE小鼠认知功能。

Objective To investigate the impact of the cannabinoid receptor agonist arachidonyl-2′-chloroethylamide(ACEA)on cognitive function in mice with sepsis-associated encephalopathy(SAE).Methods C57BL/6 mice were randomly divided into artificial cerebrospinal fluid(ACSF)and lipopolysaccharide(LPS)groups.The SAE model was established by intraventricular injection of LPS.The severity of sepsis in mice was assessed by sepsis severity score(MSS)and body mass changes.Behavioral paradigms were used to evaluate motor ability(open field test)and cognitive function(contextual fear conditioning test,Y-maze test).To evaluate the effects of ACEA intervention on SAE,mice were randomly assigned to ACSF group,ACEA intervention combined with ACSF group,LPS group,and ACEA intervention combined with LPS group.The dosage of ACEA intervention was 1.5 mg/kg.Real-time quantitative PCR was used to measure the mRNA expression levels of interleukin 1β(IL-1β),IL-6,and tumor necrosis factorα(TNF-α)in mouse hippocampal tissues.Western blot analysis was used to assess the protein levels of IL-6 and TNF-αin the hippocampus.Nissl staining was performed to examine neuronal damage in the CA1 region of the mouse hippocampus.Behavioral paradigms were again employed to evaluate motor ability and cognitive function.Results Three days after intraventricular LPS injection,mice exhibited significant cognitive dysfunction,confirming SAE modeling.Compared to the control group,the LPS group showed significant increases in mRNA of inflammatory factors such as IL-6,TNF-α,and IL-1β,together with significant increases in IL-6 and TNF-αprotein levels in the hippocampus,a decrease in Nissl bodies in the CA1 region,and significant cognitive dysfunction.Compared to the LPS group,the ACEA intervention group showed a significant decrease in the mRNA of IL-6,TNF-α,and IL-1β,a significant reduction in IL-6 and TNF-αprotein levels,an increase in Nissl bodies,and improved cognitive function.Conclusion ACEA improves cognitive function in SAE mice by inhibiting the expression levels of inflammatory factors IL-6 and TNF-α.