基于TLR-4/MyD88/NF-κB对健脾化滞丸治疗溃疡性结肠炎的机制进行拆方研究  

Splitting study of mechanism of Jianpi-Huazhi pill in treatment of ulcerative colitis based on TLR-4/MyD88/NF-κB pathway

作　　者：景姗[1] 顾玮 刘小丽 顾庆华[1] JING Shan;GU Wei;LIU Xiaoli;GU Qinghua(Department of Spleen and Stomach Diseases,Nantong Hospital Affiliated to Nanjing University of Chinese Medicine,Nantong Hospital of Traditional Chinese Medicine,Nantong 226001,China;Nantong Health Higher Vocational and Technical School,Nantong 226010,China)

机构地区：[1]南京中医药大学附属南通医院(南通市中医院)脾胃病科,南通226001 [2]南通卫生高等职业技术学校,南通226010

出　　处：《中国免疫学杂志》2024年第5期1075-1081,共7页Chinese Journal of Immunology

基　　金：江苏省“十三五”中医药重点学科建设项目(苏中医科教[2019]5号);江苏省中医药科技发展计划项目(MS2023109);南通市科技计划项目(MS12018094);南通市科技计划项目(MSZ20085);南通市“十四五”科教强卫工程(通卫科教[2021]15号)。

摘　　要：目的:通过建立脾虚湿蕴型溃疡性结肠炎(UC)大鼠模型,探讨健脾化滞丸及其不同拆方对TLR-4/MyD88/NF-κB通路的影响。方法:64只Wistar大鼠随机分为正常对照组8只、模型组56只,采用中医证候联合乙醇-2,4,6-三硝基苯磺酸灌肠法复制UC大鼠模型,造模成功后,将模型大鼠随机分为模型组(M)、美沙拉嗪组(A)、健脾化滞丸全方组(B)、健脾清化方组(C)、健脾清化活血方组(D)、健脾清化导滞方组(E)、清化导滞活血方组(F),每组8只,给予相应药物灌胃4周,疗程结束后处死所有大鼠并取材,观察大鼠体质量变化、疾病活动度及结肠病理改变,ELISA及免疫组化检测大鼠结肠组织TNF-α表达;RT-PCR及Western blot检测大鼠结肠组织TLR-4、MyD88、NF-κB mRNA及蛋白表达。结果:健脾化滞丸全方、健脾清化方、健脾清化活血方组及健脾清化导滞方组大鼠体质量明显高于模型组(P<0.01),各治疗组均能明显改善UC模型大鼠结肠组织病理损伤及疾病活动指数,其中健脾化滞丸全方组及健脾清化活血方组作用最好。各治疗组均可抑制结肠组织TLR-4、MyD88、NF-κB mRNA及蛋白表达(P<0.001),其中美沙拉嗪组、健脾化滞丸全方组、健脾清化导滞方组及清化导滞活血方组在降低NF-κB mRNA及蛋白表达上明显优于健脾清化方组及健脾清化活血方组(P<0.05)。各组均能降低结肠组织TNF-α表达(P<0.001),组间差异无统计学意义(P>0.05)。结论:健脾化滞丸及各拆方均可能通过TLR-4/MyD88/NF-κB通路发挥作用,其中黄连、煨木香、凤尾草、炮姜可能为本方核心药物,对临床具有一定指导意义。Objective:To establish ulcerative colitis(UC)rat model of spleen deficiency and dampness,and to explore effect of Jianpi-Huazhu pill and its different deconditions on TLR-4/MyD88/NF-κB pathway.Methods:A total of 64 Wistar rats were randomly divided into normal control group(n=8)and model group(n=56).UC rat model was established by traditional Chinese medicine syndrome combined with ethanol-2,4,6-trinitrobenzene sulfonic acid enema method.After successfully modeling,rats were randomly divided into model group(M),Mesalazine group(A),Jianpi-Huazhi pill group(B),Jianpi-Qinghua decoction group(C),Jianpi-Qinghua-Huoxue decoction group(D),Jianpi-Qinghua-Daozhi decoction group(E)and Qinghua-Daozhi-Huoxue decoction group(F),with 8 rats in each group,given corresponding drugs by gavage for 4 weeks.At end of course of treatment,all rats were sacrificed and samples were taken.Changes of body weight,disease activity and pathological changes of colon were observed.TNF-αexpression in colon tissue was detected by ELISA and immunohistochemistry.RT-PCR and Western blot were used to detect mRNA and protein expressions of TLR-4,MyD88 and NF-κB in rat colon tissue.Results:Rats weight in Jianpi-Huazhi pill group,Jianpi-Qinghua decoction group,Jianpi-Qinghua-Huoxue decoction group and Jianpi-Qinghua-Daozhi decoction group were obviously higher than that of model group(P<0.01),all treatment groups could obviously improve UC model rat pathological damage and disease activity index in colon tissue,and Jianpi-Huazhi pill group and Jianpi-Qinghua-Huoxue group showed better efficacy.All treatment groups could inhibit mRNA and protein expressions of TLR-4,MyD88 and NF-κB in colon tissue(P<0.001).Mesalazine group,Jianpi-Huazhi pill group,Jianpi-Qinghua-Daozhi group and Qinghua-Daozhi-Huoxue group were significantly better than Jianpi-Qinghua group and Jianpi-Qinghua-Huoxue group in reducing NF-κB mRNA and protein expressions(P<0.05).All groups could reduce expression of TNF-αin colon tissue(P<0.001),and there was no significant differenc

关 键 词：溃疡性结肠炎 健脾化滞丸 TLR-4/MyD88/NF-κB通路 TNF-α 

分 类 号：R392[医药卫生—免疫学]