【作者】 赵红伟；

【导师】 刘建平；

【作者基本信息】 河北医科大学 ， 中医内科学， 2008， 硕士

【摘要】 目的:我们认为溃疡性结肠炎(Ulcerative Colitis,UC)是以脾虚为本,湿热邪毒为标,瘀血阻络贯穿始终为病机关键。芪仙安肠方是导师在临床上治疗UC的经验方,在治疗上以“益气健脾,清热利湿,化瘀解毒,生肌敛疮”为治疗大法取得较好效果。故本研究通过动物实验,进一步观察芪仙安肠方治疗UC大鼠的疗效以及探讨本方对实验大鼠免疫机制的影响。方法:本研究采用三硝基苯磺酸(TNBS)/乙醇联合造模法诱导大鼠溃疡性结肠炎动物模型(MK),于造模后第3天分别给予芪仙安肠方大(DK)、中(ZK)、小剂量(XK)和柳氮磺胺吡啶(LK)进行灌胃治疗,观察其疗效以及探讨其作用机制。实验分三个方面研究探讨:实验一观察芪仙安肠方对溃疡性结肠炎大鼠一般状态、结肠组织病理形态学及红细胞免疫的影响;实验二芪仙安肠方对溃疡性结肠炎大鼠T细胞亚群及免疫球蛋白的影响;实验三芪仙安肠方对溃疡性结肠炎大鼠ICAM-1表达的影响。通过动物实验进一步探讨本方治疗溃疡性结肠炎的可能作用机制。结果:1实验一:1.1一般状况(DAI)与MK(2.016±0.401)比较,DK(0.429±0.153)、ZK(0.496±0.144)、XK(1.304±0.212)、LK(1.286±0.299)、BK(0.378±0.133)都能明显的降低大鼠的疾病活动指数(P<0.01);其中中药组与LK之间比较,DK、ZK明显低于LK(PDK-LK<0.01) (PZK-LK<0.01);三种不同剂量中药之间比较,DK、ZK明显低于XK(PDK-XK<0.01)(PZK-XK<0.01)。1.2结肠病理组织观察MK大鼠肠黏膜层完整,可见充血水肿和炎细胞浸润;LK和三个中药组黏膜层完整,未见充血、水肿及炎细胞浸润。1.3红细胞免疫红细胞C3b受体酵母菌花环(RBC-C3bR):与MK(4.50±1.75)比较,DK(8.5±1.58)、ZK(8.31±1.75)、BK(9.31±2.09)能非常显著增高血中红细胞C3bR受体酵母菌花环(P<0.01) ;其中中药与LK(6.49±2.41)比较, DK优于LK(PDK-LK<0.05);三种不同剂量的中药之间比较,DK与XK(6.46±2.18)比较能显著提高血中红细胞C3b受体酵母菌花环(PDK-XK<0.05)。红细胞免疫复合物花环试验(RBC-ICR):与MK(2.625±0.916)比较,DK(0.875±0.732)、ZK(0.938±0.729)、BK(0.813±0.651)能明显降低红细胞免疫复合物花环(P<0.01);其中中药与LK(1.846±0.620)之间比较,DK、ZK明显低于LK(PDK-LK<0.05) (PZK-LK<0.05);三种不同剂量的中药之间的比较,DK、ZK与XK(1.825±0.667)比较能明显降低红细胞免疫复合物花环(PDK-XK<0.05) (PZK-XK<0.05)。2实验二:2.1 T细胞亚群(%)CD3:与MK(65.98±2.54)比较,DK(52.48±4.35)、ZK(53.52±3.73)、BK(48.98±3.48)均能明显降低血中CD3含量(P<0.01);其中中药与LK(61.09±1.42)之间比较,DK、ZK血中CD3的含量能明显低于LK(PDK-LK<0.01) (PZK-LK<0.01);三种不同剂量的中药之间比较,DK、ZK明显低于XK(61.33±8.53) (PDK-XK<0.01) (PZK-XK<0.01)。CD4:与MK(51.03±3.37)比较,DK(25.24±4.40)、ZK(26.49±3.07)、XK(39.97±4.48)、LK(40.36±4.63)、BK(25.18±4.42)均能明显降低血中CD4的含量(P<0.01);其中中药组与LK之间比较,DK、ZK与LK比较能明显降低血中CD4的含量(PDK-LK<0.01) (PZK-LK<0.01);三种不同剂量的中药之间的比较,DK、ZK明显低于XK(PDK-XK<0.01) (PZK-XK<0.01)。CD8:与MK(14.69±3.45)比较,DK(27.80±7.29)、ZK(27.78±6.96)、BK(27.65±6.80)能明显升高血中CD8含量(P<0.01);其中中药与LK(21.22±6.27)之间比较,DK、ZK与LK比较具有显著性差异(PDK-LK<0.01) (PZK-LK<0.01);三种不同剂量的中药之间比较,DK、ZK与XK(21.28±6.34)比较具有显著性差异(PDK-XK<0.05) (PZK-XK<0.05)。CD4/CD8 :与MK(3.469±0.224)比较, DK(1.008±0.254)、ZK(1.034±0.245)、XK(1.869±0.043)、LK(1.868±0.129)、BK(0.991±0.191)均能明显降低血中CD4/CD8的含量(P<0.01);其中中药与LK比较,DK、ZK明显低于LK(PDK-LK<0.01) (PZK-LK<0.01);三种不同剂量的中药之间比较,DK、ZK明显低于XK(PDK-XK<0.01) (PZK-XK<0.01)。2.2免疫球蛋白IgA:与MK(0.088±0.024)比较,DK(0.036±0.017)、ZK(0.041±0.019)、BK(0.035±0.022)均能明显降低血中IgA的含量(P<0.01);其中中药与LK比较,DK、ZK明显低于LK(0.063±0.026) (PDK-LK<0.05) (PZK-LK<0.05);三种不同剂量的中药之间比较,DK、ZK与XK(0.064±0.025)比较能明降低血中IgA的含量(PDK-XK<0.05) (PZK-XK<0.05)。IgG:与MK(0.169±0.082)比较,DK(0.204±0.068)、ZK(0.210±0.085)、BK(0.169±0.082)均能降低血中IgG的含量(P<0.01);其中中药组与LK(0.300±0.105)比较,DK、ZK明显低于LK(PDK-LK<0.05) (PZK-LK<0.05);三种不同剂量的中药之间比较,DK、ZK与XK(0.300±0.100)比较能明显降低血中IgG的含量(PDK-XK<0.05) (PZK-XK<0.05)。IgM:与MK(0.309±0.070)比较,BK(0.195±0.066)能明显降低血中的IgM(P<0.01);其余各组之间比较,DK(0.271±0.056)、ZK(0.272±0.050)、XK(0.297±0.041)、LK(0.300±0.050)之间无显著性差异(P>0.05)。2.3补体C3、C4C3:与MK(0.130±0.037)比较,DK(0.209±0.038)、ZK(0.206±0.018)、BK(0.216±0.036)均能明显升高血中的补体C3的含量(P<0.01);其中中药组与LK(0.171±0.030)之间比较,DK、ZK能明显高于LK(PDK-LK<0.05) (PZK-LK<0.05);三种不同剂量的中药组之间比较,DK、ZK与XK(0.169±0.030)之间比较能明显升高血中的补体C3的含量(PDK-XK<0.05) (PZK-XK<0.05)。C4:与MK(0.024±0.011)比较,DK(0.058±0.016)、ZK(0.050±0.011)、BK(0.059±0.019)均能升高血中补体C4(P<0.01);其中中药组与LK(0.040±0.016)之间比较,DK明显高于LK(PDK-LK<0.05);三种不同剂量的中药之间比较,DK与XK(0.040±0.019)比较能明显升高血中补体C4含量(PDK-XK<0.05)。3实验三:免疫组织化学染色黏附分子ICAM-1检测与MK(5.750±1.282)比较,DK(1.875±0.991)、ZK(2.125±0.991)、XK(2.625±0.916)、LK(2.750±1.165)、BK(1.125±0.991)均能明显降低肠黏膜阳性细胞的表达(P<0.01);其余各组之间比较无显著性差异(P>0.05)。结论:芪仙安肠方能明显改善UC大鼠一般状况和结肠组织形态;提高补体C3、C4含量,减少了血液中IgG、IgA免疫反应,从而降低了ICAM-1对肠黏膜的炎症反应,IgM治疗前后变化不明显;本方还能增强UC大鼠血清中红细胞C3bR的活性,减少红细胞免疫复合物(RCIC)在肠黏膜的大量沉积,从而减轻了其对结肠黏膜上皮细胞的损伤,与模型组比较具有非常显著性差异;还能调节CD4+细胞与CD8+细胞比例,使其保持平衡,减少两者比例失衡而导致的免疫功能的紊乱。尤其是芪仙安肠方大剂量组(DK)和中剂量组(ZK)对实验大鼠的治疗作用更为明显。更多还原

【Abstract】 Objective: We also found out the main disease mechanism of UC was Pi weakness, chronic accumulation of Tanshi and further turning Shire.Qi Xian An Chang Recipe is a effective therapy, so we made up a therapy principle of healthiness spleen, dispelling damp and disappeaering carbuncle.Through the research and experiment, we could observe Qi Xian An Chang Recipe’s effects on UC and demonstrate its the immune functional mechanism.Methods: The animal experiment copied a rat UC model induced by rinitrobenzene-sulfonic acid(TNBS) and alcohol.The day after the model being made(MK), we cured them with DaJiLiang group (DK), ZhongJiLiang group(ZK), XiaoJiLiang group(Xk) and LiuDanHuangAnBiDing group(LK), observing their effects.The experiment contained four components.Experiment I: was to observe the the general state of UC rat, their pathological changes and the RBC immune;Experiment II: was about Qi Xian An Chang Recipe’s effect on T cell subsetand the immune globulin and the complement.Experiment III was about Qi Xian An Chang Recipe’s effect on the expression of ICAM-1 protein.Through these experiments we could explore UC disease mechanism further.Results:1 Experment I:1.1 The general condition (DAI)Compared with MG(2.016±0.401), DK(0.429±0.153), ZK (0.496±0.144), XK(1.304±0.212), LK(1.286±0.299)and BK (0.378±0.133)all could significant decrease the General state in UC rat (P<0.01); LK was compared with three Chinese herbal groups: DK, ZK was significant better than LK (PDK-LK<0.01) (PZK-LK<0.01); Compared among the three Chinese herbal: DK, ZK was significant better than XK (PDK-XK<0.01) (PZK-XK<0.01).1.2 Pathological damageMG rats’colonic mucosas were unabroken, with congestion, edema and infiltrating of inflammatory cells.LK and three Chinese herbal groups’colonics mucosas were unabroken, without such appearance above-mentioned.1.3 The RBC immuneRC3bR: Compared with MG(4.50±1.75), DK(8.5±1.58), ZK (8.31±1.75) and BK(9.31±2.09)all could significant increase the content of RC3bR(P<0.01);LK(6.49±2.41)was compared with three Chinese herbal groups: DK was significant increase the content of RC3bR(PDK-LK<0.05);Compared among the three Chinese herbal: DK was significant better than XK(6.46±2.18) (PDK-XK<0.05).RICR: Compared with MG(2.625±0.916), DK(0.875±0.732), ZK(0.938±0.729) and BK(0.813±0.651)all could significant decrease the content of RCIC (P<0.01);LK(1.846±0.620)was compared with three Chinese herbal groups: DK, ZK was significant decrease the content of RCIC (PDK-LK<0.05)(PZK-LK<0.05);Compared among the three Chinese herbal: DK, ZK was significant better than XK(1.825±0.667) (PDK-XK<0.05)(PZK-XK<0.05).2 Experimen II:2.1The T cell subset(%)CD3:Compared with MG(65.98±2.54),DK(52.48±4.35), ZK(53.52±3.73) and BK(48.98±3.48)all could significant decrease the percentage of CD3+(P<0.01);LK(61.09±1.42) was compared with three Chinese herbal groups: DK, ZK was significant decrease the percentage of CD3 + (PDK-LK<0.01) (PZK-LK<0.01); Compared among the three Chinese herbal: DK, ZK was significant better than XK(61.33±8.53)(PDK-XK<0.01) (PZK-XK<0.01).CD4+: Compared with MG(51.03±3.37), DK(25.24±4.40), ZK(26.49±3.07), XK(39.97±4.48), LK(40.36±4.63)and BK (25.18±4.42)all could significant decrease the percentage of CD4+ (P<0.01);LK was compared with three Chinese herbal groups: DK, ZK was significant decrease the percentage of CD4+ (PDK-LK<0.01)(PZK-LK<0.01); Compared among the three Chinese herbal: DK, ZK was significant better than XK (PDK-XK<0.01)(PZK-XK<0.01).CD8+: Compared with MG (14.69±3.45), DK(27.80±7.29), ZK(27.78±6.96)and BK(27.65±6.80)all could significant increase the percentage of CD8+ (P<0.01);LK(21.22±6.27)was compared with three Chinese herbal groups: DK, ZK was significant increase the percentage of CD8 + (PDK-LK<0.01) (PZK-LK<0.01);Compared among the three Chinese herbal: DK, ZK was significant better than XK(21.28±6.34) (PDK-XK<0.05) (PZK-XK<0.05).CD4+/CD8+: Compared with MG(3.469±0.224), DK(1.008±0.254), ZK(1.034±0.245), XK(1.869±0.043), LK(1.868±0.129) and BK(0.991±0.191)all could significant decrease the percentage of CD4+/CD8+ (P<0.01); LK was compared with three Chinese herbal groups: DK, ZK was significant decrease the percentage of CD4+/CD8+(PDK-LK<0.01) (PZK-LK<0.01); Compared among the three Chinese herbal: DK, ZK was significant better than XK(PDK-XK<0.01)(PZK-XK<0.01).2.2The immune globulinIgA: Compared with MG(0.088±0.024), DK(0.036±0.017), ZK(0.041±0.019)and BK(0.035±0.022)all could significant decrease the content of IgA (P<0.01);LK(0.063±0.026)was compared with three Chinese herbal groups: DK, ZK was significant decrease the content of IgA(PDK-LK<0.05) (PZK-LK<0.05);Compared among the three Chinese herbal: DK, ZK was significant better than XK(0.064±0.025) (PDK-XK<0.05) (PZK-XK<0.05).IgG: Compared with MG(0.169±0.082), DK(0.204±0.068), ZK(0.210±0.085)and BK(0.169±0.082)all could significant decrease the content of IgG (P<0.01);LK(0.300±0.105)was compared with three Chinese herbal groups: DK, ZK was significant decrease the content of IgG(PDK-LK<0.05) (PZK-LK<0.05);Compared among the three Chinese herbal: DK, ZK was significant better than XK(0.300±0.100)(PDK-XK<0.05) (PZK-XK<0.05).IgM: Compared with MG(0.309±0.070), BK(0.195±0.066) Could significant decrease the content of IgM (P< 0.01);Compa red among the others: DK(0.271±0.056), ZK(0.272±0.050), XK (0.297±0.041), LK(0.300±0.050)were not significant the variation( P>0.05).2.3The complement C3,C4C3: Compared with MG(0.130±0.037), DK(0.209±0.038), ZK(0.206±0.018)and BK(0.216±0.036)all could significant increase the content of C3 (P<0.01);LK(0.171±0.030)was compared with three Chinese herbal groups: DK, ZK was significant increase the content of C3(PDK-LK<0.05) (PZK-LK<0.05);Compared among the three Chinese herbal: DK, ZK was significant better than XK(0.169±0.030) (PDK-XK<0.05) (PZK-XK<0.05).C4: Compared with MG(0.024±0.011), DK(0.058±0.016), ZK(0.050±0.011)and BK(0.059±0.019)all could significant increase the content of C4 (P<0.01);LK(0.040±0.016)was compared with three Chinese herbal groups: DK was significant increase the content of C4(PDK-LK<0.05);Compared among the three Chinese herbal: DK was significant better than XK(0.040±0.019) (PDK-XK<0.05). 3 Experiment III: The expression of the Intercelluar adhesion molecule-1Compared with MG(5.750±1.282), DK(1.875±0.991), ZK (2.125±0.991), XK(2.625±0.916), LK(2.750±1.165) and BK (1.125±0.991)all could significant decrease the expression of the Intercelluar adhesion molecule-1 (P<0.01);Compared among the others were not significant the variation.( P>0.05).Conclusion:Qi Xian An Chang Recipe can improve UC rat’s general condition and colonic mucosa pathological damage;increase the content of C3, C4lessen the content of IgG, IgA in blood serum and decrease the expression of the Intercelluar adhesion molecule-1, IgM hadn’t significant variation between treating before and after.Qi Xian An Chang Recipe can improve UC rat’s the content of RC3bR in blood serum and decrease the content of RCIC in blood serum to lessen colonic mucosa damage;this Recipe also adjust the balanced between CD4+ and CD8+ and the disbalanced between CD4+and CD8+will cause immune hyperfunction.especially DK, ZK’s integrity effect is the best, significantly better than others.更多还原