摘要
目的:从Toll样受体(TLR)/髓样分化因子88(MyD88)信号通路,探讨四神丸挥发油有效治疗慢性溃疡性结肠炎的可能机制。方法:将BALB/c小鼠随机分成正常组,模型组,四神丸挥发油组,二神丸挥发油组,五味子散挥发油组和美沙拉嗪组,采用葡聚糖硫酸钠(DSS)建立慢性溃疡性结肠炎小鼠模型,灌胃给药7 d后,通过体质量、结肠质量、结肠质量指数、结肠长度、结肠镜下损伤评分评价其疗效;用酶联免疫吸附测定法(ELISA)检测结肠组织上清液中白细胞介素-4(IL-4),白细胞介素-10(IL-10),白细胞介素-17A(IL-17A),白细胞介素-21(IL-21)和γ干扰素(IFN-γ)水平,同时用蛋白免疫印迹法(Western blot)法检测各组小鼠结肠黏膜中TLR/MyD88信号通路相关蛋白,包括TLR2,MyD88,Ras相关的C3肉毒素底物1(Rac1),白细胞介素-1受体相关激酶4(IRAK4),白细胞介素-1受体相关激酶1(IRAK1),肿瘤坏死因子受体相关因子6(TRAF6),转化生长因子-β活化激酶1结合蛋白1(TAB1),转化生长因子-β活化激酶1结合蛋白2(TAB2),丝裂原活化蛋白激酶激酶6(MKK6),p38丝裂原活化蛋白激酶(p38 MAPK)和环磷腺苷效应元件结合蛋白(CREB)的表达水平。结果:与正常组比较,模型组结肠长度显著降低,结肠质量、结肠质量指数和结肠镜下病理损伤评分均显著升高,IL-10水平显著降低,IL-4,IL-21和IFN-γ水平明显升高(P<0.05,P<0.01),TLR2,MyD88,Rac1,IRAK4,IRAK1,TRAF6,TAB1,TAB2,MKK6,p38 MAPK和CREB蛋白表达量均显著升高(P<0.01);与模型组比较,经四神丸挥发油干预后,小鼠结肠长度显著增加,结肠质量、结肠质量指数和结肠镜下病理损伤评分均显著降低,且小鼠结肠组织中IL-10水平明显升高,而IL-4,IL-17A,IL-21和IFN-γ水平明显降低(P<0.05,P<0.01),同时小鼠结肠组中TLR2,MyD88,Rac1,IRAK4,IRAK1,TRAF6,TAB1,TAB2,MKK6,p38 MAPK和CREB蛋白表达量均明显下调(P<0.05,P<0.01)。结论:四神丸挥发油能有效改善慢性溃疡性结肠炎的炎性损伤,可能是通过抑制TLR/MyD88信号通路实现的。
Objective:To explore the underlying mechanism of volatile oil from Sishenwan in treating chronic ulcerative colitis through the Toll-like receptor(TLR)/myeloid differentiation factor 88(MyD88)signaling pathway.Method:The BALB/c mice were randomly divided into a normal group(normal),a model group[dextran sodium sulfate(DSS)],a Sishenwan volatile oil group,an Ershen pill volatile oil group,a Wuweizi powder volatile oil group,and a mesalazine control group.The chronic ulcerative colitis model was induced by DSS in mice.Seven days after intragastric administration,the efficacy was evaluated based on the body weight,colon weight,colon weight index,colon length,and pathological damage score under colonoscopy.The levels of interleukin(IL)-4,IL-10,IL-17A,IL-21,and interferon-γ(IFN-γ)in the supernatant of colon tissues were detected by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the expression levels of proteins related to the TLR/MyD88 signaling pathway in the colon mucosa of mice,including TLR2,MyD88,Ras-related C3 botulinum toxin substrate 1(Rac1),IL-1 receptorassociated kinase 4(IRAK4),IRAK1,tumor necrosis factor receptor(TNFR)-associated factor 6(TRAF6),transforming growth factor-β-activated kinase 1 binding protein 1(TAB1),TAB2,mitogen-activated protein kinase kinase 6 (MKK6),p38 mitogen-activated protein kinase (p38 MAPK),and cyclic adenosine monophosphate response element-binding protein(CREB).Result:Compared with the normal group,the model group showed decreased colon length,increased colon weight,colon weight index,and pathological damage score under colonoscopy,decreased IL-10 level in the colon tissues,increased IL-4,IL-17A,IL-21,and IFN-γlevels(P<0.05,P<0.01),and up-regulated protein expression of TLR2,MyD88,Rac1,IRAK4,IRAK1,TRAF6,TAB1,TAB2,MKK6,p38MAPK,and CREB(P<0.01).Compared with the model group,the Sishenwan volatile oil group showed increased colon length,reduced colon weight,colon weight index,and pathological damage score under colonoscopy,elevated IL-10 level in the colon tissues,decreased IL-4,IL-17A,IL-21,and IFN-γlevels(P<0.05,P<0.01),and down-regulated protein expression of TLR2,MyD88,Rac1,IRAK4,IRAK1,TRAF6,TAB1,TAB2,MKK6,p38MAPK,and CREB (P<0.05,P<0.01).Conclusion:The volatile oil from Sishenwan can effectively improve the inflammatory response of chronic ulcerative colitis,which may be achieved by regulating the TLR/MyD88 signaling pathway.
作者
黄佳琦
蒋青青
钟友宝
王梦雪
龙健
赵海梅
刘端勇
HUANG Jia-qi;JIANG Qin-qin;ZHONG You-bao;WANG Meng-xue;LONG Jian;ZHAO Hai-mei;LIU Duan-yong(Department of Postgraduate,Jiangxi University of Chinese Medicine,Nanchang 330004,China;Laboratory Animal Research Center for Science and Technology,Jiangxi University of Chinese Medicine,Nanchang 330004,China;College of Traditional Chinese Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004,China;Prescription and Syndrome Research Center of Jiangxi University of Chinese Medicine,Nanchang 330004,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第23期19-25,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82060838,82060799)
江西省自然科学基金项目(20202ACBL206026,20192ACB20015,20202ACBL206028,20181BAB205082)
江西省教育厅科技项目(GJJ196049)
江西省研究生创新专项(研字[2020]39号)。
