摘要目的 探讨卡巴胆碱对高氧诱导小鼠急性肺损伤(hyperoxia-induced acute lung injury,HALI)的保护作用及其相关机制.方法 健康雄性ICR小鼠32只,随机(随机数字法)分为对照组、高氧暴露3 d组(HO3d组)、高氧暴露3 d+卡巴胆碱组(HO3d+Carba组)、卡巴胆碱组(Carba组),每组8只.各组小鼠造模完成后均在光镜下观察肺组织病理学改变;采用Western-blot检测肺组织Toll样受体4(Toll-like receptor 4,TLR4)及核因子-κB(nuclear factor-kappa B,NF-κB)的表达;采用实时定量PCR(RT-PCR)检测肺组织高迁移率族蛋白B1(high mobility group box-1 protein,HMGB-1)及肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)mRNA的表达.计量资料两两比较采用LSD-t检验,多组间比较采用单因素方差分析,以P<0.05为差异有统计学意义.结果 对照组和Carba组比较,各指标差异均无统计学意义(P>0.05),肺组织结构无明显异常损伤改变;HO3d组小鼠肺组织TLR4、NF-κB蛋白表达量以及HMGB-1、TNF-αmRNA表达量明显高于对照组(P<0.01),且肺组织表面有明显出血,病理损伤严重;HO3d+Carba组小鼠肺组织TLR4、NF-κB蛋白表达量以及肺组织HMGB-1、TNF-αmRNA较HO3d组明显降低(P<0.01),同时肺组织损伤程度也较HO3d组减轻.结论 高氧可通过上调肺组织TLR4、NF-κB的表达,引起肺组织炎性损伤;卡巴胆碱可以减少高氧肺损伤HMGB-1、TNF-α 炎症因子的释放,其机制可能与抑制TLR4/NF-κB信号通路相关,对HALI有一定保护作用.

abstractsObjective To investigate the protective effect of Carbachol on hyperoxia-induced acute lung injury (HALI) in mice and its related mechanisms. Methods Thirty-two healthy male ICR mice were randomly divided into four groups:control group, hyperoxia exposure three days group (HO3d group), hyperoxia exposure three days + Carbachol group (HO3d+Carba group), and Carbachol group (Carba group), eight mice in each group. The pathological changes of lung tissue in each group were observed under light microscope after the models were completed in each group.The expression of TLR4 and NF-κB protein in lung tissues were detected by Western blot, and the expression of HMGB-1 and TNF-α mRNA in lung tissues by RT-PCR. LSD-t test was used for sample pairwise comparison, and one-way ANOVA for intergroup comparison. P<0.05 was considered statistically significant. Results There was no statistical difference between the control group and the Carba group (P> 0.05), and no obvious abnormal changes in lung tissue structure. The expression of TLR4, NF-κB protein and HMGB-1 and TNF-α mRNA in the HO3d group were significantly higher than those in the control group (P<0.01), and there were obvious bleeding on the surface of the lung tissue and severe pathological damage. The&nbsp;expression of TLR4,NF-κB protein and HMGB-1 and TNF-α mRNA in the HO3d+Carba group were significantly lower than those in the HO3d group(P<0.01), while lung tissue damage degree was also lower than that in the HO3d group. Conclusions Hyperoxia can increase the expression of TLR4 and NF-κB in lung tissues, and cause inflammatory injury in lung tissue. Carbachol can reduce the release of HMGB-1 and TNF-α inflammatory factors in hyperoxia-induced acute lung injury, and its mechanism is related to the inhibition of TLR4/NF-κB signal pathway, which has a protective effect on HALI.