摘要目的 观察卡巴胆碱对脓毒症大鼠肺微血管内皮细胞(VEC)功能的影响.方法 将SPF级健康雄性SD大鼠30只随机分成三组:氯化钠注射液组(SHAM组)、脓毒症组(LPS组)、卡巴胆碱组(CBL组).LPS组经左侧股静脉注射脂多糖(LPS)10 mg/kg,SHAM组经左侧股静脉注射等量氯化钠注射液,卡巴胆碱组在注射LPS后注射卡巴胆碱10μg/kg.分别在各组模型制备完毕后6 h经腹主动脉取血用ELISA法检查肿瘤坏死因子-α(TNF-ɑ)水平,取右肺下叶肺组织用Real-time PCR检测SSeCKS的mRNA表达,Western blot检测各组右肺下叶肺组织E-选择素蛋白质表达水平情况,取右肺中叶行透射电镜检查.结果 SHAM组表达少量的TNF-ɑ、SSeCKS、E-选择素,LPS能够上调他们的表达,CBL组TNF-ɑ、SSeCKS、E-选择素较LPS组表达降低(P<0.05).透射电镜结果显示CBL组肺VEC水肿和凋亡等表现较LPS组相对减轻.结论 卡巴胆碱不仅可以有效抑制脓毒症大鼠血浆TNF-ɑ水平,抑制炎症反应,而且能下调肺微血管内皮细胞SSeCKS和E-选择素的表达,并减轻LPS所致的肺微血管内皮细胞超微结构改变.提示卡巴胆碱能够改善脓毒症大鼠肺微血管内皮细胞的通透性.

abstractsObjective To investigate the effects of carbachol on pulmonary microvascular endo-thelial cell function in septic rats .Methods Thirty healthy male SPF Sprague-Dawley rats were random-ly divided into three groups:saline group ( SHAME ) , lipopolysaccharide group ( LPS ) and carbachol group ( CBL) .Sepsis model was reproduced by the left femoral vein sublethal dose of lipopolysaceharide ( LPS,10 mg/kg ) .Carbachol group was injected with carbachol ( 10 μg/kg ) after injection of LPS . Tumor necrosis factor-a ( TNF-ɑ) in the blood from the abdominal aorta were measured by ELISA ( en-zyme-linked immuno sorbent assay ) method, SSeCKS from lower lobe of right lung were measured by Real-time PCR method , E-selectin from lower lobe of right lung were measured by Western blot method after injection 6 h.The middle lobe of right lung was taken for transmission electron microscope examina-tion after injection 6 h.Results The TNF-ɑ, SSeCKS and E-selectin in SHAM had low expression , but LPS can make their expression increased .The expression of TNF-ɑ, SSeCKS mRNA, and E-selectin pro-tein in CLB group was lower than those in the LPS group ( P<0.05 ) .The results of transmission electron microscopy showed that VEC edema and apoptosis in CBL group relatively reduced in LPS group . Conclusions Carbachol can not only effectively inhibite the level of TNF-ɑin septic rats plasma , inhibit inflammation, but also down-regulate the pulmonary microvascular endothelial cells with SSeCKS and E-selectin expression , and reduce the changes of ultrastructure of VEC , indicating that carbachol can im-prove the permeability of pulmonary microvascular endothelial cell in septic rats .