四磨汤通过调控NF-κB信号通路改善重度脓毒症大鼠结肠上皮细胞屏障功能障碍炎症状态作用研究  

Effect of Simo decoction on inflammation of colonic epithelial cell barrier dysfunction in rats with severe sepsis via NF-κB signal pathway

作　　者：温剑艺[1] 王首红[1] 刘艳红[1] 张崇健[1] 郭伟新[1] WEN Jianyi;WANG Shouhong;LIU Yanhong;ZHANG Chongjian;GUO Weixin(Guangdong Provincial People’s Hospital,Guangzhou 510080,China)

机构地区：[1]广东省人民医院广东省医学科学院,广东广州510080

出　　处：《陕西中医》2023年第10期1365-1371,共7页Shaanxi Journal of Traditional Chinese Medicine

基　　金：广东省中医药局科研项目(20221012)。

摘　　要：目的:探讨四磨汤对重度脓毒症大鼠结肠上皮细胞屏障功能障碍炎症状态的改善作用以及其对核转录因子κB(NF-κB)信号的调控机制。方法:采用盲肠结扎穿孔术(CLP)构建大鼠重度脓毒症模型;实验分组为正常组、模型组、四磨汤低剂量组、四磨汤高剂量组、激活剂组,每组10只;试剂盒检测各组大鼠血清中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的含量;末端脱氧核苷酸转移酶(TdT)介导的脱氧尿苷三磷酸(dUTP)缺口末端标记(TUNEL)染色检测各组大鼠结肠组织的细胞凋亡;免疫荧光检测结肠组织中高迁移率蛋白-1(HMGB1)、核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)的表达;免疫组化检测结肠中白细胞共同抗原(CD45)、闭合蛋白1(Claudin-1)和闭锁蛋白1(ZO-1)的表达;Western blot法检测结肠组织中NF-κB、磷酸化NF-κB65(NF-κB p65)、B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)的表达。结果:四磨汤能明显下调重度脓毒症大鼠血清中TNF-α和IL-6的含量;抑制结肠组织中的细胞凋亡,降低结肠组织中HMGB1和NLRP3的表达,上调结肠组织中CD45、ZO-1和Claudin-1的表达;抑制NF-κB p65和Bax的表达,升高Bcl-2的表达,差异有统计学意义(均P<0.05)。结论:四磨汤能明显抑制重度脓毒症大鼠结肠上皮细胞的炎症应激状态,缓解结肠组织的病理损伤,改善结肠黏膜的屏障功能,这可能与抑制NF-κB信号的磷酸化有关。Objective:To investigate the effect of Simo decoction on the inflammatory state of colonic epithelial cell barrier dysfunction in rats with severe sepsis and its regulation mechanism on nuclear factor-κB(NF-κB)signaling pathway.Methods:Cecal ligation and puncture(CLP)was used to establish the rat model of severe sepsis.The experimental groups were divided into normal group,model group,low dose Simo decoction group,high dose Simo decoction group and activator group,with 10 rats in each group.The kit was used to detect the content of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in serum.TUNEL staining was used to detect the apoptosis in colon tissues.Immunofluorescence was used to detect the expression of high mobility group box-1 protein(HMGB1)and nucleotide binding oligomerization domain-like receptor protein 3(NLRP3)in colon tissues.Immunohistochemistry was used to detect the expression of common antigen(CD45),closure protein 1(Claudin-1)and zonula occludens-1(ZO-1)in colon tissues.Western blot was used to detect the expression of NF-κB,phosphorylated NF-κB 65(NF-κB p65),B cell lymphoma-2(Bcl-2),and Bcl-2 associated x protein(Bax).Results:Simo decoction could significantly down-regulate the contents of TNF-αand IL-6 in serum of rats with severe sepsis,inhibit the apoptosis in colon,decrease the expression of HMGB1 and NLRP3,up-regulate the expression of CD45,ZO-1 and Claudin-1,inhibit the expression of NF-κB p65 and Bax,and increase the expression of Bcl-2,and the differences were statistically significant(all P<0.05).Conclusion:Simo decoction can obviously inhibit the inflammatory stress state of colonic epithelial cells in rats with severe sepsis,alleviate the pathological injury of colonic tissue and improve the barrier function of colonic mucosa,which may be related to the phosphorylated inhibition of NF-κB signaling pathway.

关 键 词：重度脓毒症 四磨汤 结肠上皮细胞屏障 炎症状态 核转录因子κB信号 

分 类 号：R749.13[医药卫生—神经病学与精神病学]