白术多糖通过调控TLR4/NF⁃κB信号通路对重症急性胰腺炎大鼠肠黏膜免疫屏障的影响  

Effects of polysaccharide of Atractylodes macrocephala Koidz on intestinal mucosal immune barrier in rats with severe acute pancreatitis via TLR4/NF⁃κB signaling pathway

作　　者：江勇 朱大侠 刘礼剑 JIANG Yong;ZHU Da-xia;LIU Li-jian(Emergency Department of Nanhua Hospital Affiliated to Nanhua University,Hengyang 421002,China)

机构地区：[1]南华大学附属南华医院急诊科,湖南衡阳421002

出　　处：《中成药》2021年第3期624-629,共6页Chinese Traditional Patent Medicine

摘　　要：目的探讨白术多糖对重症急性胰腺炎(SAP)大鼠肠黏膜免疫屏障的保护作用及其机制研究。方法采用逆行胆胰管注射5%牛磺胆酸钠溶液法建立SAP模型,术后立即给予相应药物处理。于术后48 h,检测血清中淀粉酶、TNF⁃α(肿瘤坏死因子⁃α)、白介素IL⁃6和IL⁃1β水平;HE染色观察胰腺和小肠组织病理学变化;流式细胞术检测小肠黏膜中CD4^(+)T细胞和CD8^(+)T细胞亚群比例;放射免疫法检测小肠黏膜中sIgA(分泌性免疫球蛋白A)水平;Western blot检测小肠组织中TLR4/NF⁃κB通路相关蛋白表达。结果与假手术组比较,SAP模型组大鼠胰腺和小肠组织损伤严重,血清淀粉酶活性及TNF⁃α、IL⁃6和IL⁃1β水平增加,小肠黏膜CD4+T细胞亚群比例及CD4^(+)/CD8^(+)比值和sIgA水平降低,而CD8+T细胞亚群比例上升,同时小肠组织中MyD88(髓样分化因子)和TLR4蛋白及NF⁃κB p65蛋白磷酸化水平增加(均P<0.05)。与SAP模型组比较,白术多糖高剂量组和乌司他丁组大鼠胰腺和小肠组织损伤有所改善,血清淀粉酶活性和TNF⁃α、IL⁃6、IL⁃1β水平降低,小肠黏膜中CD4^(+)T细胞亚群比例及CD4^(+)/CD8^(+)比值和sIgA水平增加,CD8^(+)T细胞亚群比例降低,而小肠组织中MyD88、TLR4蛋白及NF⁃κB p65磷酸化水平降低(均P<0.05)。结论白术多糖可改善重症急性胰腺炎大鼠肠黏膜免疫屏障损伤,其机制可能与抑制TLR4/NF⁃κB信号通路活化有关。AIM To investigate the protective effect of polysaccharide of Atractylodes macrocephala Koidz(PAM)on intestinal mucosal immune barrier in rats with severe acute pancreatitis(SAP)and its mechanism.METHODS Sixty male SD rats were randomly divided into sham operation group,model group(SAP),positive drug ulinastatin group(2×10^(4) U·kg^(-1)),low⁃dose PAM group(200 mg·kg^(-1))and high⁃dose PAM group(400 mg·kg^(-1)),with 12 rats in each group.The SAP rat models established with 5%sodium taurine injection into retrograde cholangiopancreatic tube were treated with the corresponding drug immediately after operation.Forty⁃eight h after the operation,rats were subjected to the detection of serum levels of amylase,TNF⁃α,IL⁃6 and IL⁃1β;the observation of pathological changes of pancreas and small intestine by HE staining;the determination of the subsets of CD4^(+)T cells and CD8+T cells in intestinal mucosa by flow cytometry;the detection of sIgA level in intestinal mucosa by radioimmunoassay;and the identification of expression of TLR4/NF⁃κB pathway related protein by Western blot.RESULTS Compared with the sham operation group,the SAP group shared severely damaged pancreatic and intestinal tissues;increased serum levels of amylase,TNF⁃α,IL⁃6 and IL⁃1β;decreased proportion of CD4^(+)T cell subsets,CD4^(+)/CD8^(+)ratio and sIgA level;increased proportion of CD8^(+)T cell subsets;and improved intestinal levels of MyD88,TLR4 and phosphorylation of NF⁃κBp65 protein(all P<0.05).In contrast to the SAP group,both high⁃dose PAM group and ulinastatin group demonstrated improvement in terms of pancreatic and intestinal tissue damage,.decreased serum levels of amylase,TNF⁃α,IL⁃6 and IL⁃1β,increased proportion of CD4^(+)T cell subsets,CD4^(+)/CD8^(+)ratio and sIgA level,decreased proportion of CD8^(+)T cell subsets,decreased intestinal levels of MyD88,TLR4 and phosphorylation of NF⁃κBp65 protein(all P<0.05).CONCLUSION PAM improves the damage of intestinal mucosal immune barrier in SAP rats pos

关 键 词：白术多糖 重症急性胰腺炎 肠黏膜免疫屏障 TLR4/NF⁃κB 

分 类 号：R285.5[医药卫生—中药学]