大麻二酚对脓毒症模型小鼠生存率和炎症反应的影响

Effect of cannabidiol on survival rate and inflammatory response of septic mice

背景寻找能够治疗并缓解脓毒症症状的药物一直是临床迫切需要解决的问题。大麻二酚(cannabidiol,CBD)有希望被应用于多种神经退行性疾病和免疫相关疾病的治疗中,本课题组将目光放在大麻二酚对于脓毒症的治疗上。目的观察大麻二酚对脓毒症小鼠生存率、炎症细胞因子水平[白细胞介素-1b(interleukin-1,IL-1b)、γ干扰素(interferon-γ,IFN-γ)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和IL-6]、肺组织病理改变和免疫细胞浸润情况的影响。方法本试验分为两部分,共使用72只小鼠。第一部分为生存率观察实验,第二部分为标本采集实验。实验一:选取36只小鼠随机分为3组,每组12只,分别为模型对照组、大麻二酚溶剂对照组和大麻二酚实验组,盲肠结扎穿孔术(cecal ligation and puncture,CLP)后分别于12 h、24 h、36 h、48 h、60 h、72 h腹腔注射1 mL的0.9%氯化钠注射液、大麻二酚溶剂或20 mg/kg的CBD溶液。观察各组小鼠在CLP术后72 h生存率;实验二:将剩余36只小鼠按照上述方案分组,在CLP术后12 h、24 h、36 h腹腔注射1 mL的0.9%氯化钠注射液、溶剂或20 mg/kg的CBD溶液,于术后48 h处死小鼠,检测炎症细胞因子水平(IL-1b、IFN-γ、TNF-α和IL-6)、肺部病理改变和肝免疫细胞浸润情况。结果相较于模型组和大麻二酚溶剂对照组,大麻二酚实验组小鼠的细胞因子水平(IL-1b、IFN-γ、TNF-α和IL-6)降低(P<0.01);CD4^(+)、CD8^(+)T细胞、自然杀伤细胞在脓毒症小鼠肝中聚集情况明显减少,肺病理损伤改善;模型对照组、大麻二酚溶剂对照组、大麻二酚实验组小鼠72 h生存率分别为16.7%、25%、58.3%。结论CBD可以减轻小鼠脓毒症肺损伤,抑制炎性细胞因子的分泌,减少免疫细胞浸润,提高脓毒症小鼠的生存率。

Background Pursuing drugs that can treat and relieve the symptoms of sepsis has been an urgent clinical problem.Cannabidiol is promising to be used in the treatment of a variety of neurodegenerative diseases and immune-related diseases.Our research group focuses on the treatment of sepsis by cannabidiol.Objective To observe the effects of cannabidiol(CBD)on the survival rate,inflammatory cytokine levels(IL-1b,IFN-γ,TNF-αand IL-6),lung pathological changes and immune cell infiltration in sepsis mice.Methods This experiment was divided into two parts,with 72 mice in total.The first part was the survival rate observation experiment,and the second part was the specimen collection experiment.Thirty-six mice were selected in the first experiment and randomly divided into model control group,solvent control group and cannabidiol experimental group,with 12 mice in each group.At 12 h,24 h,36 h,48 h,60 h,and 72 h after CLP,1 mL of 0.9%sodium chloride aqueous solution,cannabidiol solvent or 20 mg/kg CBD solution was intraperitoneally injected.The survival rate of 72 h after cecal ligation and perforation was observed.The remaining 36 mice were included in the second experiment and grouped according to the above scheme,and 1 mL of 0.9%sodium chloride aqueous solution,solvent or 20 mg/kg CBD solution was intraperitoneally injected at 12 h,24 h,and 36 h after CLP.The mice were sacrificed at 48 h after CLP,the levels of inflammatory cytokines(IL-1b,IFN-γ,TNF-αand IL-6),lung pathological changes and liver immune cell infiltration were detected.Results Compared with the model control group and the solvent control group,the cytokine levels of IL-1b,IFN-γ,TNF-αand IL-6 in the CBD experimental group decreased significantly(all P<0.01).The accumulation of CD4^(+)T,CD8^(+)T cells and natural killer cells in the septic liver were reduced,thus improving the lung pathological damage.The 72 h survival rate of mice in the model control group,solvent control group and CBD experimental group was 16.7%,25%and 58.3%respectively.Conclusion The above results indicate that CBD can reduce lung injury in mice with sepsis,inhibit the secretion of inflammatory cytokines,reduce immune cell infiltration and prolong the survival of septic mice.