Objective To explore the mechanism of butylphthalide (NBP) in regulating microglia activation and inflammatory cytokine expression in the hippocampus of the mouse model of delayed encephalopathy after carbon monoxide poisoning (DEACMP). Methods Wild-type C57 adult mice with normal cognitive function were selected,and DEACMP was modeled by static inhalation of carbon monoxide.The mice were randomized into three groups:DEACMP,control,and NBP.The NBP group was administrated with NBP suspension at 6 mg/kg by gavage for 21 days,and the DEACMP and control groups were administrated with the same amount of vegetable oil by gavage.The hippocampal injury was observed by HE staining.The protein level of ionized calcium-binding adapter molecule 1 (IBA1) was determined by Western blotting,and the levels of downstream inflammatory cytokines were measured by ELISA. Results Compared with the control group,the DEACMP and NBP groups showed prolonged escape latency (P=0.001,P=0.029),reduced nerve cells (P=0.001,P=0.035),up-regulated expression of IBA1 (P=0.001,P=0.042),increased mean fluorescence intensity of IBA1 (P=0.001,P=0.021),and elevated levels of tumor necrosis factor-α (TNF-α) (P=0.002,P=0.024),interleukin (IL)-6 (P=0.001,P=0.015),and IL-1β (P=0.001,P=0.023).Compared with the DEACMP group,the NBP group showed shortened escape latency (P=0.025),increased nerve cells (P=0.039),down-regulated expression of IBA1 (P=0.035),decreased average fluorescence intensity of IBA1 (P=0.031),and lowered levels of TNF-α (P=0.028),IL-6 (P=0.037),and IL-1β (P=0.034). Conclusion NBP can inhibit the activation of microglia and reduce the expression of inflammatory factors,thereby alleviating cognitive dysfunction and brain tissue damage caused by DEACMP.
目的 探究丁苯酞(NBP)对一氧化碳中毒迟发性脑病(DEACMP)小鼠海马组织内小胶质细胞活化及炎症因子表达的作用机制。方法 筛选认知功能正常的C57成年野生型小鼠,采用静态吸入一氧化碳法制备DEACMP小鼠模型,并随机分为3组:DEACMP组、对照组及NBP组,其中,NBP组灌胃NBP混悬液6 mg/kg连续21 d,DEACMP组和对照组灌胃等量植物油。采用HE染色观察小鼠海马组织损伤情况,Western blot法测定离子钙结合衔接分子1(IBA1)蛋白表达水平,ELISA法检测下游炎症因子表达水平。结果 与对照组比较,DEACMP组和NBP组逃避潜伏期显著延长(P=0.001,P=0.029),神经细胞数量明显减少(P=0.001,P=0.035),IBA1表达显著增加(P=0.001,P=0.042),IBA1平均荧光强度显著增强(P=0.001,P=0.021),肿瘤坏死因子-α(TNF-α)(P=0.002,P=0.024)、白细胞介素(IL)-6(P=0.001,P=0.015)、IL-1β(P=0.001,P=0.023)表达水平显著增高。与DEACMP组比较,NBP组逃避潜伏期缩短(P=0.025),神经细胞数量增多(P=0.039),IBA1表达降低(P=0.035),IBA1平均荧光强度减弱(P=0.031),TNF-α(P=0.028)、IL-6(P=0.037)、IL-1β(P=0.034)表达水平降低。结论 NBP可通过抑制小胶质细胞活化减少炎症因子的表达,进而减轻DEACMP导致的认知功能障碍及脑组织损伤。.
Keywords: butylphthalide; carbon monoxide poisoning; delayed encephalopathy; inflammatory cytokines; microglia.