枸杞多糖抑制溃疡性结肠炎小鼠结肠组织中干扰素-γ的表达
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摘要
目的观察枸杞多糖对慢性溃疡性结肠炎小鼠结肠组织中干扰素-γ(IFN-γ)表达的影响。方法小鼠设正常组、DSS组和枸杞多糖组,DSS组循环饮用1.5%DSS,枸杞多糖组在DSS组的基础上灌胃给予200mg·(kg·d)-1的枸杞多糖,记录各组小鼠的体重变化,HE染色观察各组小鼠结肠组织病理变化,并通过q-PCR和ELISA检测三组小鼠结肠组织内IFN-γmRNA和蛋白的表达水平。结果 DSS组小鼠自第7天起各时点体重均低于正常组(P<0.05);正常组小鼠结肠黏膜厚薄均匀,隐窝结构规则,无炎症细胞浸润,DSS组小鼠结肠黏膜厚薄不均,隐窝出现分支、萎缩、扭曲等结构改变,肠壁内有大量淋巴细胞、浆细胞浸润,枸杞多糖组小鼠结肠黏膜结构破坏有所减轻,肠壁内炎症细胞浸润减少;正常组IFN-γmRNA的相对表达量为(0.85±0.15),DSS组为(1.11±0.54),差异无统计学意义(P>0.05),枸杞多糖组的相对表达量为(0.22±0.11)低于DSS组(P<0.05);正常组小鼠结肠组织匀浆液中IFN-γ蛋白的浓度为(65.95±0.08)pg·mL-1,枸杞多糖组IFN-γ的浓度为(92.21±15.36)pg·mL-1均低于DSS组(115.39±9.62)pg·mL-1(P均<0.05)。结论枸杞多糖下调DSS小鼠结肠组织中IFN-γ的表达,可为枸杞多糖治疗溃疡性结肠炎的研究奠定基础。
Objective To observe the effect of Lycium barbarum polysaccharide(LBP)on expression of IFN-γin the colon tissue of mice with chronic ulcerative colitis. Methods Mice were randomly divided into three groups: control group,DSS group and LBP group. The DSS group was given 1.5% DSS for 4 cycles,and the LBP group was given 200 mg·(kg·d)-1 LBP on the basis of DSS group. Weight changes of mice in each group were recorded. HE staining was used to observe the pathological changes of colon tissue in each group. The levels of IFN-γmRNA and protein in colon tissues were detected by q-PCR and ELISA. Results The weight of mice in the normal group increased with the growth. The weight of the mice in the DSS group was lower than that in the normal group from the 7 th day(P<0.05). There was no significant difference in the weight change between the mice in the LBP group and the DSS group(P>0.05). The colonic mucosa of the normal group was uniform in thickness,the structure of the crypt was regular,and there was no infiltration of inflammatory cells.The thickness of the colonic mucosa in the DSS group was uneven,and the crypts showed branching,atrophy,distortion and other structural changes. There were a large number of lymphocytes and plasma cells in the intestinal mucosa. The destruction of colonic mucosal structure in the LBP group was alleviated,and the inflammatory cell in the intestinal mucosa was reduced. The relative expression of IFN-γ mRNA in the normal group was(0.85 ±0.15),and(1.11±0.54)in the DSS group,the difference was not statistically significant(P>0.05). Compared with the DSS group,the relative expression of IFN-γ in the LBP group was(0.22±0.11),the difference was statistically significant(P<0.05). The concentration of IFN-γ protein in the colonic homogenate of the normal group was(65.95±0.08)pg·m L-1,and that of the DSS group was(115.39±9.62)pg·m L-1. The difference was statistically significant(P<0.05). Compared with the DSS group,the concentration of IFN-γ in the LBP group was(92.21 ±15.36)pg·m L-1,the difference was statistically significant(P<0.05).Conclusion LBP down-regulates the expression of IFN-γ in colon tissue of DSS mice,which lays a foundation for the research of LBP treatment for ulcerative colitis.
Objective To observe the effect of Lycium barbarum polysaccharide(LBP)on expression of IFN-γin the colon tissue of mice with chronic ulcerative colitis. Methods Mice were randomly divided into three groups: control group,DSS group and LBP group. The DSS group was given 1.5% DSS for 4 cycles,and the LBP group was given 200 mg·(kg·d)-1 LBP on the basis of DSS group. Weight changes of mice in each group were recorded. HE staining was used to observe the pathological changes of colon tissue in each group. The levels of IFN-γmRNA and protein in colon tissues were detected by q-PCR and ELISA. Results The weight of mice in the normal group increased with the growth. The weight of the mice in the DSS group was lower than that in the normal group from the 7 th day(P<0.05). There was no significant difference in the weight change between the mice in the LBP group and the DSS group(P>0.05). The colonic mucosa of the normal group was uniform in thickness,the structure of the crypt was regular,and there was no infiltration of inflammatory cells.The thickness of the colonic mucosa in the DSS group was uneven,and the crypts showed branching,atrophy,distortion and other structural changes. There were a large number of lymphocytes and plasma cells in the intestinal mucosa. The destruction of colonic mucosal structure in the LBP group was alleviated,and the inflammatory cell in the intestinal mucosa was reduced. The relative expression of IFN-γ mRNA in the normal group was(0.85 ±0.15),and(1.11±0.54)in the DSS group,the difference was not statistically significant(P>0.05). Compared with the DSS group,the relative expression of IFN-γ in the LBP group was(0.22±0.11),the difference was statistically significant(P<0.05). The concentration of IFN-γ protein in the colonic homogenate of the normal group was(65.95±0.08)pg·m L-1,and that of the DSS group was(115.39±9.62)pg·m L-1. The difference was statistically significant(P<0.05). Compared with the DSS group,the concentration of IFN-γ in the LBP group was(92.21 ±15.36)pg·m L-1,the difference was statistically significant(P<0.05).Conclusion LBP down-regulates the expression of IFN-γ in colon tissue of DSS mice,which lays a foundation for the research of LBP treatment for ulcerative colitis.
引文
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[13]Globig AM,Hennecke N,Martin B,et al.Comprehensive intestinal T helper cell profiling reveals specific accumulation of IFN-γ+IL-17+coproducing CD4+T cells in active inflammatory bowel disease[J].Inflamm Bowel Dis,2014,20(12):2321-2329.
[14]Lim BO.Coriolus versicolor suppresses inflammatory bowel disease by inhibiting the expression of STAT1and STAT6 associated with IFN-γand IL-4 expression[J].Phytother Res,2011,25(8):1257-1261.
[2]胡品津,钱家鸣,吴开春,等.我国炎症性肠病诊断与治疗的共识意见(2012年·广州)[J].内科理论与实践,2013(1):61-75.
[3]Guo BJ,Bian ZX,Qiu HC,et al.Biological and clinical implications of herbal medicine and natural products for the treatment of inflammatory bowel disease[J].Ann N Y Acad Sci,2017,1401(1):37-48.
[4]Chan HC,Ng SC.Emerging biologics in inflammatory bowel disease[J].J Gastroenterol,2017,52(2):141-150.
[5]Solina G,MandalàS,La Barbera C,et al.Current management of intestinal bowel disease:the role of surgery[J].Updates Surg,2016,68(1):13-23.
[6]Li M,Gao J,Tang Y,et al.Traditional herbal medicinederived sulforaphene LFS-01 reverses colitis in mice by selectively altering the gut microbiota and promoting intestinal Gamma-delta T cells[J].Front Pharmacol,2018,8:959.
[7]Cheng J,Zhou ZW,Sheng HP,et al.An evidencebased update on the pharmacological activities and possible molecular targets of Lycium barbarum polysaccharides[J].Drug Des Devel Ther,2014,9:33-78.
[8]Ananthakrishnan AN.Epidemiology and risk factors for IBD[J].Nature Reviews Gastroenterology&Hepatology,2015,12(4):205-217.
[9]Katsanos KH,Papadakis KA.Inflammatory bowel disease:updates on molecular targets for biologics[J].Gut Liver,2017,11(4):455-463.
[10]Park JH,Peyrin-Biroulet L,Eisenhut M,et al.IBDimmunopathogenesis:A comprehensive review of inflammatory molecules[J].Autoimmun Rev,2017,16(4):416-426.
[11]Kmiec'Z,Cyman M,'Slebioda TJ.Cells of the innate and adaptive immunity and their interactions in inflammatory bowel disease[J].Adv Med Sci,2017,62(1):1-16.
[12]Nakanishi Y,Sato T,Takahashi K,et al.IFN-γ-dependent epigenetic regulation instructs colitogenic monocyte/macrophage lineage differentiation in vivo[J].Mucosal Immunol,2018,11(3):871-880.
[13]Globig AM,Hennecke N,Martin B,et al.Comprehensive intestinal T helper cell profiling reveals specific accumulation of IFN-γ+IL-17+coproducing CD4+T cells in active inflammatory bowel disease[J].Inflamm Bowel Dis,2014,20(12):2321-2329.
[14]Lim BO.Coriolus versicolor suppresses inflammatory bowel disease by inhibiting the expression of STAT1and STAT6 associated with IFN-γand IL-4 expression[J].Phytother Res,2011,25(8):1257-1261.