通腑理肺汤对脓毒症大鼠肠黏膜免疫屏障保护作用的实验研究  

Study on Protective Effect of Tongfu Lifei Decoction(通腑理肺汤)on Immune Barrier of Intestinal Mucosa in Rats with Sepsis

作　　者：陈辉珍[1] 高健婷 黄小哲 CHEN Huizhen;GAO Jianting;HUANG Xiaozhe(Hangzhou Hospital of Chinese Medicine,Hangzhou,Zhejiang 310007)

机构地区：[1]杭州市中医院,浙江杭州310007

出　　处：《中国中医药科技》2024年第5期776-781,共6页Chinese Journal of Traditional Medical Science and Technology

基　　金：浙江省杭州市科技发展计划项目(20201203B181)。

摘　　要：目的:探究通腑理肺汤(Tongfu Lifei Decoction,TLD)对脓毒症大鼠肠黏膜免疫屏障的保护作用及其机制。方法:通过盲肠结扎穿刺术(cecal ligation puncture,CLP)制备大鼠脓毒症模型,随机分为模型组(Model)、通腑理肺汤低剂量组(3.6 g/kg,TLD-L)、通腑理肺汤高剂量组(7.2 g/kg,TLD-H),同时设假手术组(Sham)作为对照,分组给药,1次/日,连续给药7 d。ELISA检测血清IL-1β、TNF-α含量,流式细胞仪检测脾脏细胞Th17、Treg细胞百分比变化,HE染色观察肠组织病理变化,qPCR检测肠组织caspase-3(胱天蛋白酶3)和caspase-9(胱天蛋白酶3)mRNA的表达水平。Western blot检测肠组织caspase-3和caspase-9蛋白表达水平。结果:与Sham组相比,Model组大鼠血清IL-1β、TNF-α含量、脾脏细胞Th17、Treg细胞数量极显著升高(P<0.01)。与Model组相比,通腑理肺汤组大鼠血清IL-1β、TNF-α含量、Th17细胞数目极显著降低(P<0.01),Treg细胞数量显著升高(P<0.05)。HE染色结果显示通腑理肺汤可减少炎症细胞浸润,改善肠黏膜损伤。qPCR检测结果显示通腑理肺汤可下调caspase-3、caspase-9 mRNA表达水平。Western blot检测结果显示通腑理肺汤可下调caspase-3、caspase-9蛋白表达水平。结论:通腑理肺汤可保护脓毒症大鼠肠黏膜免疫屏障,其可能是通过维持Th17/Treg平衡、下调IL-1β、TNF-α含量及caspase-3、caspase-9的mRNA和蛋白表达水平发挥肠黏膜免疫屏障保护作用。Objective:Exploring the protective effect of Tongfu Lifei Decoction(TLD)on the immune ba-rrier of intestinal mucosal of rats with sepsis and its mechanism.Methods:A rat sepsis model was prepared by cecal ligation puncture(CLP)surgery,then these rats was randomly divided into model group(Model),low-dose Tongfu Lifei Decoction group(3.6 g/kg,TLD-L)and high-dose Tongfu Lifei Decoction group(7.2 g/kg,TLD-H),sham operation group(Sham)was set as the control group.After 7 days of drug administration,serum IL-1βand TNF-αlevels were determined by ELISA and changes in the percentage of Th17 and Treg cells in spleen cells were detected by flow cytometry.HE staining was performed to observe the pathological changes of intestinal tissue.qPCR was performed to detect the expression levels of caspase-3 and caspase-9 mRNA in intestinal tissues.Western blot was performed to detect the expression levels of caspase-3 and caspase-9 proteins in intestinal tissues.Results:Compared with the sham group,the serum IL-1βand TNF-αlevels,splenocyte Th17 and Treg cell numbers were significantly higher in the model group(P<0.01).Compared with the Model group,IL-1β,TNF-αlevels and Th17 cells were significantly lower(P<0.01)and Treg cells were significantly higher(P<0.05)in the Tongfu Lifei Decoction group.HE staining showed that Tongfu Lifei Decoction could reduce inflammatory cells infiltration and improve the intestinal mucosal injury.qPCR showed that Tongfu Lifei Decoction could down-regulate caspase-3 and caspase-9 mRNA expression levels.Western blot showed that Tongfu Lifei Decoction could down-regulate caspase-3 and caspase-9 protein expression levels.Conclusion:Tongfu Lifei Decoction can protect the intestinal mucosal immune barrier in rats with sepsis,probably by maintaining Th17/Treg balance,down-regulating IL-1β,TNF-αand caspase-3 and caspase-9 mRNA and protein expression levels.

关 键 词：通腑理肺汤 脓毒症 肠黏膜免疫屏障 TH17/TREG IL-1Β TNF-α caspase-3 CASPASE-9 大鼠 

分 类 号：R285.5[医药卫生—中药学]