目的:观察钩吻素子(koumine，KM)对脂多糖(lipopolysaccharide，LPS)诱导的小鼠脓毒症及其所致急性肝损伤的治疗作用，并探究可能的作用机制。方法:48只C57 BL/6小鼠随机分为对照组、模型组、10 mg·kg-1 MCC950组和KM 7。5，15，30 mg·kg-1剂量组，每组8只。各组小鼠一次性腹腔注射相应药物或溶剂(生理盐水)，给药1 h后，对照组小鼠给予等体积生理盐水，其余各组小鼠给予LPS 1 mg·kg-1腹腔注射，诱导建立小鼠脓毒症模型。LPS干预4 h后，采用全自动生化分析仪检测血清谷草转氨酶(aspartate ami-notransferase，AST)和谷丙转氨酶(alanine transaminase，ALT)活性;HE染色法观察肝脏组织病理学;ELISA法检测血清和腹腔灌洗液白细胞介素-1β(interleukin-1β，IL-1β)水平;RT-qPCR法检测肝脏组织中核苷酸结合寡聚化结构域样受体3(nucleotide binding oligomerization domain like receptor protein 3，NLRP3)、半胱氨酸天冬氨酸蛋白酶-1前体(cysteine aspartic protease-1 precursor，pro-caspase-1)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD，ASC)、IL-1β前体(interleukin-1βprecursor，pro-IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)和白细胞介素-6(interleukin-6，IL-6)mRNA水平。结果:KM显著降低脓毒症小鼠血清中AST和ALT水平，显著降低小鼠血清及腹腔灌洗液中IL-1β水平;肝脏病理切片显示KM治疗后小鼠组织受损程度较模型组稍有减轻;RT-qPCR研究结果证实，KM显著降低炎性因子TNF-α和IL-6 mRNA的表达水平，并能显著下调模型小鼠肝脏组织NLRP3，pro-caspase-1，ASC和pro-IL-1βmRNA的表达水平。结论:KM对脓毒症致小鼠急性肝损伤有治疗作用，其机制可能与抑制NLRP3炎症小体的活化及相关促炎细胞因子的释放有关。
The therapeutic effect of koumine on acute liver injury in mice with sepsis and its mechanisms
Objective:To explore the therapeutic effect and possible mechanisms of koumine (KM) on lipopolysaccharide (LPS)-induced sepsis and acute liver injury in mice.Methods:A total of 48 C57BL/6 mice were randomly divided into control group,model group,MCC95010 mg·kg-1 group and KM 7.5,15,30 mg·kg-1 groups,with eight mice in each group.Mice were given a one-time intraperitoneal injection of the corresponding drugs or an equal volume of normal saline.1 h later,the mice in the control group were given equal volume of normal saline,and those in the other groups were given 1 mg·kg-1 LPS intraperitoneally to construct an LPS-induced sepsis mouse model.4 h after LPS administration,serum aspartate aminotransferase ( AST ) and alanine transaminase ( ALT ) activities were detected by automatic biochemical analyzer.The serum AST and ALT activities were determined by an automatic biochemical analyzer.The levels of interleukin-1β in serum and peritoneal lavage fluid were measured by an ELISA kit.RT-qPCR was used to detect the mRNA levels of nucleotide binding oligomerization domain like receptor protein 3 (NLRP3),cysteine aspartic protease-1 precursor (pro-caspase-1),apoptosis-associated speck-like protein containing a CARD ( ASC),IL-1β precursor ( pro-IL-1β),tumor necrosis factor-α ( TNF-α) and IL-6 in liver tissue.Results:KM significantly decreased the serum levels of AST and ALT in sepsis mice and markedly decreased the levels of IL-1β in serum and peritoneal lavage fluid.The pathological section of liver showed that the degree of tissue damage after KM treatment was slightly reduced than that in the model group.RT-qPCR results confirmed that KM significantly decreased the mRNA expression levels of inflammatory factors TNF-α and IL-6,and significantly decreased the mRNA expression levels of NLRP3 inflammator-related proteins including NLRP3,pro-caspase-1,ASC and pro-IL-1β in liver tissue of model mice.Conclusion:KM shows a significant therapeutic effect on acute liver injury in mice with sepsis,and its mechanisms may be attributed to inhibiting the activation of NLRP3 inflammasome and the release of related pro-inflammatory cytokines.
kouminesepsisliver injurynucleotide binding oligomerization domain like receptor protein 3interleukin-1β
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维普
