Objective:To investigate the effects of Sishen pill on the expression of type I interferon(IFN)and type III interferon and their receptors in colonic tissues of mice with acute ulcerative colitis(UC).Methods:Male C57B...Objective:To investigate the effects of Sishen pill on the expression of type I interferon(IFN)and type III interferon and their receptors in colonic tissues of mice with acute ulcerative colitis(UC).Methods:Male C57BL/6Cnc mice were randomly divided into control group,model group,sishenwan group and salazosulfapyridine group.The model was made with 0.2 mL 4%dextran sodium sulfate(DSS)for 5 days,and the control group was given 0.2mL normal saline by gavage.On the second day of modeling,sishen pill group was given 0.2mL 1.5 g·kg^(-1) sishen pill,and SASP group was given 0.2mL 0.25 g·kg^(-1) sulfasalazine,twice a day,for 7 days.During the administration period,the disease activity index(DAI)of mice was calculated every day.After administration,the histopathological changes of colon tissues of mice in each group were observed by hematoxylin eosin(HE)staining,and the histological scores were calculated.The expression of IFN-α,IFN-β,IFN-λ2 and IFN-λ3 mRNAs in colon tissues of mice in each group were detected by qRT-PCR.The expression levels of IFN-α,IFN-β,IFN-λ2 and IFN-λ3 in colon tissues of mice in each group were detected by ELISA.Western blot was used to detect the expression of interferon receptors IFNAR1,IFNAR2 and IFNLR1 in colon tissues of mice in each group.Results:Compared with the control group,the DAI of mice increased significantly(P<0.001)in the model group.The inflammatory cells in colonic tissues infiltrated heavily,lymph nodes enlarged,colonic mucosal structure destroyed,crypt structure lost,inflammation involved a wide range,and the histological score increased significantly(P<0.001).The levels of IFN-α,IFN-βand IFNλ2 mRNA were significantly decreased(P<0.05,P<0.05,P<0.01).The expression levels of IFN-α,IFN-β,IFN-λ2 and IFN-λ3 were significantly decreased(P<0.01,P<0.01,P<0.001,P<0.001).The levels of IFNAR1,IFNAR2 and IFNLR1 were significantly decreased(P<0.01,P<0.05,P<0.01).Compared with the model group,the DAI decreased significantly(P<0.001)in Sishen pill group,the infiltration of inflammatory cells in colon tissue were significantly reduced,the structural regeneration of colon mucosa was significantly recovered,the crypt structure was significantly recovered,the lymph nodes were significantly reduced,the range of inflammation involvement was reduced,and the histological score was significantly reduced(P<0.001).The levels of IFN-α,IFN-βand IFN-λ2 mRNA were significantly increased(P<0.01,P<0.001,P<0.001).The levels of IFN-α,IFN-β,IFN-λ2 and IFN-λ3 were significantly increased(P<0.01,P<0.001,P<0.01,P<0.001).The levels of IFNAR1,IFNAR2 and IFNLR1 were significantly increased(P<0.05,P<0.001,P<0.05).Conclusion:Sishen pill may alleviate the symptoms and signs of mice with acute ulcerative colitis by regulating the expression of type I and type III interferon and their receptors in colon tissues.展开更多
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
基金National Natural Science Foundation of China(No.82160835)。
文摘Objective:To investigate the effects of Sishen pill on the expression of type I interferon(IFN)and type III interferon and their receptors in colonic tissues of mice with acute ulcerative colitis(UC).Methods:Male C57BL/6Cnc mice were randomly divided into control group,model group,sishenwan group and salazosulfapyridine group.The model was made with 0.2 mL 4%dextran sodium sulfate(DSS)for 5 days,and the control group was given 0.2mL normal saline by gavage.On the second day of modeling,sishen pill group was given 0.2mL 1.5 g·kg^(-1) sishen pill,and SASP group was given 0.2mL 0.25 g·kg^(-1) sulfasalazine,twice a day,for 7 days.During the administration period,the disease activity index(DAI)of mice was calculated every day.After administration,the histopathological changes of colon tissues of mice in each group were observed by hematoxylin eosin(HE)staining,and the histological scores were calculated.The expression of IFN-α,IFN-β,IFN-λ2 and IFN-λ3 mRNAs in colon tissues of mice in each group were detected by qRT-PCR.The expression levels of IFN-α,IFN-β,IFN-λ2 and IFN-λ3 in colon tissues of mice in each group were detected by ELISA.Western blot was used to detect the expression of interferon receptors IFNAR1,IFNAR2 and IFNLR1 in colon tissues of mice in each group.Results:Compared with the control group,the DAI of mice increased significantly(P<0.001)in the model group.The inflammatory cells in colonic tissues infiltrated heavily,lymph nodes enlarged,colonic mucosal structure destroyed,crypt structure lost,inflammation involved a wide range,and the histological score increased significantly(P<0.001).The levels of IFN-α,IFN-βand IFNλ2 mRNA were significantly decreased(P<0.05,P<0.05,P<0.01).The expression levels of IFN-α,IFN-β,IFN-λ2 and IFN-λ3 were significantly decreased(P<0.01,P<0.01,P<0.001,P<0.001).The levels of IFNAR1,IFNAR2 and IFNLR1 were significantly decreased(P<0.01,P<0.05,P<0.01).Compared with the model group,the DAI decreased significantly(P<0.001)in Sishen pill group,the infiltration of inflammatory cells in colon tissue were significantly reduced,the structural regeneration of colon mucosa was significantly recovered,the crypt structure was significantly recovered,the lymph nodes were significantly reduced,the range of inflammation involvement was reduced,and the histological score was significantly reduced(P<0.001).The levels of IFN-α,IFN-βand IFN-λ2 mRNA were significantly increased(P<0.01,P<0.001,P<0.001).The levels of IFN-α,IFN-β,IFN-λ2 and IFN-λ3 were significantly increased(P<0.01,P<0.001,P<0.01,P<0.001).The levels of IFNAR1,IFNAR2 and IFNLR1 were significantly increased(P<0.05,P<0.001,P<0.05).Conclusion:Sishen pill may alleviate the symptoms and signs of mice with acute ulcerative colitis by regulating the expression of type I and type III interferon and their receptors in colon tissues.
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.
基金supported by the Major Program of the National Natural Science Foundation of China (grant nos.82192910 and 82192911)the Innovation Team and Talents Cultivation Program of the National Administration of Traditional Chinese Medicine (grant nos.ZYYCXTD-D-202207 and ZYYCXTD-C-202009)。
文摘溃疡性结肠炎(ulcerative colitis,UC)是一种病因复杂且复发率高的慢性非特异性炎症性肠病。《2017年溃疡性结肠炎中西医结合诊疗共识意见》中将出自《证治准绳》的四神丸作为治疗脾肾阳虚型UC的基础中药方剂。以四神丸为主方加减化裁、联合其他中药复方、联合西药以及其他中医特色疗法治疗慢性复发性UC具有疗效确切、不良反应小、复发率低、多靶点及多通路调控等优势。调节免疫是四神丸治疗UC的主要作用机制,主要表现在对记忆性T细胞(Memory T cell,TM)、辅助型T细胞1(T helper 1 cell,Th1)/辅助型T细胞2(T helper 2 cell,Th2)、调节性T细胞(Treg)、辅助性T细胞17(T helper cell 17,Th17)、滤泡辅助性T细胞(follicular B helper T cells,Tfh)/滤泡调节性T细胞(follicu⁃lar regulatory T cells,Tfr)、树突状细胞、B淋巴细胞和肠道菌群等以及多条信号通路调控作用,显示出四神丸良好的免疫药理作用。因此,就近年来四神丸治疗UC的临床运用及免疫药理学研究展开综述,以期为中医药治疗UC提供更多思路和途径。
