CD40在溃疡性结肠炎发病机制中作用的实验研究

发布时间：2018-04-13 07:12

本文选题：CD40 + 溃疡性结肠炎　；参考：《苏州大学》2014年博士论文

【摘要】：第一部分CD40在溃疡性结肠炎组织中的表达及意义目的：研究CD40分子在溃疡性结肠炎组织中的表达，及其与溃疡性结肠炎严重度之间的相关性。方法：应用免疫组化技术检测56例溃疡性结肠炎组织和10例正常组织中CD40分子表达。结果：CD40分子在溃疡性结肠炎组织中的阳性表达率为62.5%，明显高于对照组（P＜0.05）。与轻度溃疡性结肠炎相比，CD40阳性表达率在中度溃疡性结肠炎中明显升高（P＜0.05）。结论：溃疡性结肠炎患者结肠组织中CD40表达异常。第二部分抗CD40单克隆抗体在溃疡性结肠炎中治疗效应及作用机制的实验研究目的初步探讨抗CD40单克隆抗体在葡聚糖硫酸钠(dextran sulfate sodium, DSS)诱导的小鼠溃疡性结肠炎(ulcerative colitis, UC)中的治疗效应及作用机制。方法24只BALB/c小鼠按随机数字表分为：A组(对照组，n=6)、B组(模型组，n=6)、C组(奥沙拉秦干预组，n=6)、D组(抗CD40单克隆抗体干预组，n=6)。A组小鼠自由饮用蒸馏水7天；B、C、D组小鼠自由饮用5%DSS溶液7天以制成UC小鼠模型；造模的同时于第1、3、5及7天分别给以A、B组腹腔注射无菌蒸馏水。C组小鼠每天灌服奥沙拉秦(60mg/kg)、D组小鼠于造模第1、3、5、7天腹腔注射抗CD40单克隆抗体（20ug/次），，于造模第8天，处死所有各组小鼠。大体观察各组小鼠疾病活动指数(disease activity index, DAI)，常规病理切片评估结肠组织病理学评分(histopathological score, HPS)。RT-PCR测定各组小鼠远端结肠中IFN-γmRNA、TNF-αmRNA的表达，ELISA法测定各组小鼠血清中IFN-γ、TNF-α含量的变化。结果(1)与对照组DAI(0.29±0.48)和HPS(0.16±0.32)相比，模型组小鼠DAI(6.21±1.25)和HPS(7.00±0.58)明显增高(p0.05)；与模型组相比，奥沙拉秦干预组C组DAI(4.58±2.36)和HPS(4.35±0.76)均有不同程度的下降(p0.05)；抗CD40单克隆抗体干预组D组DAI(3.98±1.86)和HPS(3.25±0.66)亦有不同程度的下降(p0.05)(2)模型组IFN-γ mRNA、TNF-mRNA表达量（分别为3.21±0.26、1.23±0.36）明显高于对照组(分别为1.35±0.18、0.21±0.13)(p0.05)，奥沙拉秦干预后，IFN-γ mRNA、TNF-mRNA表达在C组(2.58±0.24、0.72±0.15)、抗CD40单克隆抗体干预组D组(2.34±0.25、0.59±0.18)均显著下降(p0.05)。（3）对照组小鼠血清中IFN-γ、TNF-α的水平分别为(215.18±32.51)、(20.06±5.20)，模型组小鼠血清中IFN-γ、TNF-α的表达比例分别为(953.32±40.96)、(42.97±8.76)，模型组小鼠血清中IFN-γ、TNF-α的表达比例与对照组相比明显升高(P0.05)，奥沙拉秦干预后C组、抗CD40抗体干预组D组小鼠血清中IFN-γ、TNF-α的表达比例分别为C组(768.73±31.86)、(37.75±6.75)、D组(693.47±51.38)、(30.93±3.18)，与模型组相比干预组均有明显降低(P0.05)；结论(1)抗CD40抗体干预组干预的UC小鼠模型大体症状可明显好转，结肠炎症程度亦得到明显减轻；(2)奥沙拉秦干预后C组、抗CD40抗体干预组D组的UC小鼠远端结肠中IFN-γ mRNA、TNF-αmRNA明显下降，提示促炎性细胞因子在UC发病中可能发挥重要作用；(3)抗CD40抗体干预的UC小鼠血清中IFN-γ和TNF-α的表达水平均明显下降；提示抗CD40单克隆抗体可下调效应细胞因子，减轻UC炎症。
[Abstract]:Expression and significance of first part CD40 in ulcerative colitis

Objective : To study the expression of CD40 molecule in ulcerative colitis and its correlation with the severity of ulcerative colitis .

Methods : Immunohistochemical technique was used to detect the expression of CD40 in 56 ulcerative colitis and 10 normal tissues .

Results : The positive expression rate of CD40 molecule in ulcerative colitis was 62.5 % , which was significantly higher than that in control group ( P < 0.05 ) .

Conclusion : The expression of CD40 in colonic tissues of patients with ulcerative colitis is abnormal .

Effect of the second part of anti - CD40 monoclonal antibody on ulcerative colitis and its mechanism of action

experimental study

Objective To investigate the therapeutic effect and mechanism of anti - CD40 monoclonal antibody in ulcerative colitis ( UC ) induced by dextran sulfate sodium ( DSS ) . Methods Twenty - four BALB / c mice were randomly divided into group A ( control group , n = 6 ) , group B ( model group , n = 6 ) , group C ( group C , n = 6 ) and D group ( anti - CD40 monoclonal antibody intervention group , n = 6 ) . Group A mice were free to drink distilled water for 7 days ;
B , C and D groups were free to drink 5 % DSS solution for 7 days to prepare UC mice model ;
All groups of mice were sacrificed on Days 1 , 3 , 5 and 7 , respectively , and the mice were given intraperitoneal injection of the anti - CD40 monoclonal antibody ( 20ug / kg ) on day 1 , 3 , 5 and 7 . All the groups of mice were sacrificed on the 8th day of the molding . The disease activity index ( DAI ) and the pathological score of colonic tissues were assessed by routine pathological sections . The expression of IFN - 纬 mRNA and TNF - 伪 mRNA in the distal colon of each group were determined by RT - PCR . The changes of IFN - 纬 and TNF - 伪 in serum of each group were determined by ELISA .

Results ( 1 ) Compared with control group DAI ( 0.29 卤 0.48 ) and hps ( 0.16 卤 0.32 ) , DAI ( 6.21 卤 1.25 ) and hps ( 7.00 卤 0.58 ) in model group were significantly higher ( p < 0.05 ) .
Compared with model group , DAI ( 4.58 卤 2.36 ) and HPs ( 4.35 卤 0.76 ) decreased in group C ( p < 0.05 ) .
The levels of IFN - 纬 and TNF - 伪 in the serum of the model group were significantly higher than those in the control group ( 2.58 卤 0 . 24 , 0 . 21 卤 0 . 13 ) ( P < 0 . 05 ) . The levels of IFN - 纬 and TNF - 伪 in the serum of the model group were significantly higher than those in the control group ( 2.58 卤 0 . 24 , 0 . 21 卤 0 . 15 ) , and the levels of IFN - 纬 and TNF - 伪 in the serum of the model group were significantly lower than those in the control group ( P < 0 . 05 ) .

Conclusion ( 1 ) The general symptoms of UC mice with anti - CD40 antibody intervention group can be improved obviously , and the degree of colonic inflammation is obviously reduced .
( 2 ) There was a significant decrease in IFN - 纬 mRNA and TNF - 伪 mRNA in the distal colon of UC mice in group D of anti - CD40 antibody , suggesting that pro - inflammatory cytokines might play an important role in the pathogenesis of UC .
( 3 ) The levels of IFN - 纬 and TNF - 伪 in serum of UC mice with anti - CD40 antibody were significantly decreased .
It is suggested that anti - CD40 monoclonal antibody can down - regulate effector cytokines and reduce UC inflammation .

【学位授予单位】：苏州大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R574.62

【参考文献】