OBJECTIVE:To investigate the therapeutic effect of Sishen Wan(四神丸,SSW)on ulcerative colitis(UC)induced by dinitrobenzene sulfonic acid and its effect on toll-like receptor 2/interleukin-1 receptor-associated kinase...OBJECTIVE:To investigate the therapeutic effect of Sishen Wan(四神丸,SSW)on ulcerative colitis(UC)induced by dinitrobenzene sulfonic acid and its effect on toll-like receptor 2/interleukin-1 receptor-associated kinase-4/nuclear factor-κB(TLR2/IRAK4/NF-κB)signaling pathway in colonic tissue.METHODS:In this study,120 Sprague–Dawley rats were randomly divided into blank and model groups.The experimental UC model in rats was established by subcutaneous injection of hydrocortisone+senna gavage for 21 d+dinitrobenzene sulfonic acid(DNBS)/ethanol solution enema.The successful model rats were randomly divided into the model group;mesalazine(0.36 g/kg)group;and high-,medium-,and low-dose SSW(24,12,and 6 g/kg)groups.The model and blank groups were gavaged with equal volumes of distilled water once a day for 21 d.The general condition of the rats was observed,and the body mass,fecal properties,and occult blood were recorded for calculating the disease activity index(DAI)score.The colonic tissue of the rats was collected,and its general morphology and pathological form were noted for obtaining the colonic mucosal injury index(CMDI)score.Hematoxylin-eosin staining was used to view the pathological changes of the colon tissue in each group,apoptosis of the cells was detected using terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling staining,and quantitative real-time polymerase chain reaction was used to measure the expressions of TLR2,myeloid differentiation primary response gene 88(My D88),IRAK4,and NF-κB p65 mRNA in the colon tissue.The expressions of TLR2,My D88,IRAK4,and NF-κB p65 protein were detected using western blotting and immunohistochemistry assay,and the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the colon tissue were determined using enzyme linked immunosorbent assay.RESULTS:Compared with the blank group,the general condition of the model group was relatively poor.The DAI and CMDI scores of the model group increased significantly(P<0.01),the glands and intestinal mucosa disappeared partially,and several inflammatory cells infiltrated and gathered in the mucosal layer and base layer of the rats in the model group.Furthermore,the cell apoptosis and expression levels of TLR2,My D88,IRAK4,and NF-κB p65 mRNA and protein in the colon tissue of rats in the model group increased significantly(P<0.01).The levels of IL-1βand TNF-αincreased significantly in the colon tissue of rats in the model group(P<0.01).After treatment with SSW,compared with the model group,the general condition of the UC rats improved.Moreover,the DAI and CMDI scores of the UC rats decreased significantly(P<0.05),and the pathological changes in the colon tissue of the UC rats tended to be normal.The cell apoptosis and expression levels of TLR2,My D88,IRAK4,and NF-κB p65 mRNA and protein in the colon tissue of the UC rats decreased gradually(P<0.01),and the levels of IL-1βand TNF-αdecreased significantly(P<0.01).CONCLUSION:SSW can improve the general condition and alleviate the intestinal mucosal injury of UC model rats.Additionally,SSW can inhibit the TLR2/IRAK4/NF-κB signaling pathway,but further studies are required to confirm it.展开更多
目的:运用网络药理学方法及分子对接模拟技术,分析四神丸治疗腹泻型肠易激综合征的作用机制。方法:采用网络药理学方法,基于中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis ...目的:运用网络药理学方法及分子对接模拟技术,分析四神丸治疗腹泻型肠易激综合征的作用机制。方法:采用网络药理学方法,基于中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)数据库、中药分子机制的生物信息学分析工具(BATMAN-TCM)数据库及对相关文献的检索,找出四神丸中各药物活性成分及相互作用靶点,并以口服生物利用度≥30%,类药性≥0.18为条件进行筛选;采用Cytoscape 3.8.2软件绘制药物-活性成分-靶点的可视化网络;利用GeneCards数据库检索腹泻型肠易激综合征相关靶点;通过VENNY网络平台将四神丸与腹泻型肠易激综合征的潜在靶点做韦恩图;利用STRING数据库构建蛋白质-蛋白质相互作用网络模型,并通过Cytoscape 3.8.2软件进行优化;基于Matescape数据平台,对潜在靶点进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)通路富集分析,最后通过Autodock进行分子对接检验。结果:筛选后得出活性成分51个、靶点270个,腹泻型肠易激综合征靶点976个,四神丸治疗腹泻型肠易激综合征的靶点116个,包括肿瘤坏死因子(Tumor Necrosis Factor,TNF)、丝氨酸/苏氨酸蛋白激酶1(Akt Serine/Threonine Kinase 1,AKT1)、转录因子JUN、细胞肿瘤抗原p53(Cellular Tumor Antigen p53,TP53)、白细胞介素-6(Interleukin 6,IL-6)等。KEGG富集分析主要得到癌症通路、动脉粥样硬化信号通路、丙型病毒性肝炎、催乳素信号通路等。分子对接结果显示核心成分与核心靶点的结合活性稳定。结论:四神丸治疗腹泻型肠易激综合征是通过其多成分、多靶点、多通路的特点实现的,为进一步的临床应用及实验研究提供了理论依据。展开更多
基金Supported by the National Natural Science Foundation of China:to Explore the Immune Mechanism of Treating Ulcerative Colitis by Regulating Treg/Th17 Cell Imbalance by Warming the Kidney and Invigorating the Spleen through Intestinal Flora(No.81960826)
文摘OBJECTIVE:To investigate the therapeutic effect of Sishen Wan(四神丸,SSW)on ulcerative colitis(UC)induced by dinitrobenzene sulfonic acid and its effect on toll-like receptor 2/interleukin-1 receptor-associated kinase-4/nuclear factor-κB(TLR2/IRAK4/NF-κB)signaling pathway in colonic tissue.METHODS:In this study,120 Sprague–Dawley rats were randomly divided into blank and model groups.The experimental UC model in rats was established by subcutaneous injection of hydrocortisone+senna gavage for 21 d+dinitrobenzene sulfonic acid(DNBS)/ethanol solution enema.The successful model rats were randomly divided into the model group;mesalazine(0.36 g/kg)group;and high-,medium-,and low-dose SSW(24,12,and 6 g/kg)groups.The model and blank groups were gavaged with equal volumes of distilled water once a day for 21 d.The general condition of the rats was observed,and the body mass,fecal properties,and occult blood were recorded for calculating the disease activity index(DAI)score.The colonic tissue of the rats was collected,and its general morphology and pathological form were noted for obtaining the colonic mucosal injury index(CMDI)score.Hematoxylin-eosin staining was used to view the pathological changes of the colon tissue in each group,apoptosis of the cells was detected using terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling staining,and quantitative real-time polymerase chain reaction was used to measure the expressions of TLR2,myeloid differentiation primary response gene 88(My D88),IRAK4,and NF-κB p65 mRNA in the colon tissue.The expressions of TLR2,My D88,IRAK4,and NF-κB p65 protein were detected using western blotting and immunohistochemistry assay,and the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the colon tissue were determined using enzyme linked immunosorbent assay.RESULTS:Compared with the blank group,the general condition of the model group was relatively poor.The DAI and CMDI scores of the model group increased significantly(P<0.01),the glands and intestinal mucosa disappeared partially,and several inflammatory cells infiltrated and gathered in the mucosal layer and base layer of the rats in the model group.Furthermore,the cell apoptosis and expression levels of TLR2,My D88,IRAK4,and NF-κB p65 mRNA and protein in the colon tissue of rats in the model group increased significantly(P<0.01).The levels of IL-1βand TNF-αincreased significantly in the colon tissue of rats in the model group(P<0.01).After treatment with SSW,compared with the model group,the general condition of the UC rats improved.Moreover,the DAI and CMDI scores of the UC rats decreased significantly(P<0.05),and the pathological changes in the colon tissue of the UC rats tended to be normal.The cell apoptosis and expression levels of TLR2,My D88,IRAK4,and NF-κB p65 mRNA and protein in the colon tissue of the UC rats decreased gradually(P<0.01),and the levels of IL-1βand TNF-αdecreased significantly(P<0.01).CONCLUSION:SSW can improve the general condition and alleviate the intestinal mucosal injury of UC model rats.Additionally,SSW can inhibit the TLR2/IRAK4/NF-κB signaling pathway,but further studies are required to confirm it.
文摘目的:运用网络药理学方法及分子对接模拟技术,分析四神丸治疗腹泻型肠易激综合征的作用机制。方法:采用网络药理学方法,基于中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)数据库、中药分子机制的生物信息学分析工具(BATMAN-TCM)数据库及对相关文献的检索,找出四神丸中各药物活性成分及相互作用靶点,并以口服生物利用度≥30%,类药性≥0.18为条件进行筛选;采用Cytoscape 3.8.2软件绘制药物-活性成分-靶点的可视化网络;利用GeneCards数据库检索腹泻型肠易激综合征相关靶点;通过VENNY网络平台将四神丸与腹泻型肠易激综合征的潜在靶点做韦恩图;利用STRING数据库构建蛋白质-蛋白质相互作用网络模型,并通过Cytoscape 3.8.2软件进行优化;基于Matescape数据平台,对潜在靶点进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)通路富集分析,最后通过Autodock进行分子对接检验。结果:筛选后得出活性成分51个、靶点270个,腹泻型肠易激综合征靶点976个,四神丸治疗腹泻型肠易激综合征的靶点116个,包括肿瘤坏死因子(Tumor Necrosis Factor,TNF)、丝氨酸/苏氨酸蛋白激酶1(Akt Serine/Threonine Kinase 1,AKT1)、转录因子JUN、细胞肿瘤抗原p53(Cellular Tumor Antigen p53,TP53)、白细胞介素-6(Interleukin 6,IL-6)等。KEGG富集分析主要得到癌症通路、动脉粥样硬化信号通路、丙型病毒性肝炎、催乳素信号通路等。分子对接结果显示核心成分与核心靶点的结合活性稳定。结论:四神丸治疗腹泻型肠易激综合征是通过其多成分、多靶点、多通路的特点实现的,为进一步的临床应用及实验研究提供了理论依据。
