基因静默对肠易激综合征内脏高敏感大鼠肠道树突状细胞免疫功能的影响  

Effects of gene silence on immune function of colonic dendritic cells in rats with irritable bowel syndrome visceral hypersensitivity

作　　者：庄肇朦[1] 张璐[1] 陶丽媛[1] 李蒙[1] 吕宾[1] 

机构地区：[1]浙江中医药大学附属第一医院消化科,杭州310000

出　　处：《中华消化杂志》2016年第2期96-100,共5页Chinese Journal of Digestion

基　　金：国家自然科学基金（H0307）

摘　　要：目的了解PDIA3基因在IBS内脏高敏感大鼠结肠黏膜异常免疫应答中的作用及机制。方法将48只成年SD大鼠分为空白对照组、空病毒组（IBS-1）、PDIA3基因静默组（IBS-2）和模型对照组（IBS-3），每组各12只。免疫组织化学法检测回盲部结肠组织表面分子CD103的表达，流式分选技术分离肠系膜淋巴结树突状细胞（DC），磁珠分选技术分离脾脏CD4^＋/CD8^＋T淋巴细胞，DC和CD4^＋/CD8^＋T淋巴细胞共培养，MTT法检测DC促淋巴细胞增殖能力，ELISA法检测DC促CD4^＋/CD8^＋T淋巴细胞分泌IL-4、IL-9水平。统计学处理采用独立样本t检验。结果大鼠肠道CD103标记的DC计数在空白对照组为（6.25±1.14）个/高倍镜视野（HPF），低于IBS-3组的（10.83±1.03）个/HPF（t＝10.07，P〈0.05）；IBS-2组为（7.42±0.90）个/HPF，低于IBS-3组，差异有统计学意义（t＝－9.25，P〈0.05）。空白对照组和IBS-3组大鼠DC促进CD4^＋T淋巴细胞增殖的MTT值分别为0.54±0.01和0.60±0.01（t＝3.373，P〈0.05）；IBS-2组为0.53±0.01，低于IBS-3组，差异有统计学意义（t＝－3.139，P〈0.05）；DC促进CD8^＋T淋巴细胞增殖的MTT值分别为0.52±0.01和0.59±0.00（t＝3.539，P〈0.01）；IBS-2组为0.54±0.01，低于IBS-3组，差异有统计学意义（t＝－3.183，P〈0.01）。空白对照组和IBS-3组大鼠DC促CD4^＋T淋巴细胞分泌IL-4水平分别为10.24±0.09和16.61±1.00（t＝3.222，P〈0.05）；IBS-2组为11.75±0.54，低于IBS-3组，差异有统计学意义（t＝－3.539，P〈0.01）。空白对照组和IBS-3组大鼠DC促CD4^＋T淋巴细胞分泌IL-9水平分别为15.86±10.19和43.51±11.32（t＝4.529，P〈0.05）；IBS-2组为29.05±2.09，低于IBS-3组，差异有统计学意义（t＝－6.841，P〈0.01）。空白对照组和IBS-3组大鼠DC促CD8^＋T淋巴细胞分泌IL-4水平分别为7.35±0.12和13.91±0.57（t＝19.557，P〈0.01）；IBS-2组为8.63±0.24，低于IBS-3组，差异�Objective To explore the role and mechanism of protein disulfide isomerase A3 （PDIA3） in abnormal colonic mucosa immune response of irritable bowel syndrome （IBS） rats with visceral hypersensitivity. Methods A total of 48 SD rats were divided into blank control group （n= 12）, empty virus group （IBS-1） （n= 12）, PDIA3 silencing group （IBS-2） （n= 12） and model control group（IBS-3） （n= 12）. The expression of CD103 in ileocecus colonic tissues was detected by immunohistochemistry. Dendritic ceils （DC） of mesenteric lymph node were isolated by flow cytometry. CD4^＋/CD8^＋ T cells of spleen were separated by immune-magnetic beads sorting technique. DC and CD4^＋/CD8^＋ T cells were co-cultured. The proliferation ability of lymphocytes promoted by DC was measured by methyl thiazolyl tetrazolium （MTT）. The secretion levels of interleukin （IL）- 4 and IL-9 of CD4^＋/CD8^＋ T cells stimulated by DC were determined by enzyme linked immunosorbent assay （ELISA） method. Independent sample t-test was performed for statistical analysis. Results The cell number of CD103 positive DC of rats colon of blank control group, IBS-3 group was 6.25± 1.14 and 10.83 ±1.03 （t = 10.07,P〈0.05）; that of IBS-2 group was 7. 42±0.90, and compared with that of IBS-3 group, the difference was statistically significant （t= -9. 25, P 〈 0.05）. The MTT value of CD4^＋ T cells proliferation stimulated by DC of blank control group and IBS-3 group was 0.54±0.01 and 0. 60_--4-0. 01 （t=3. 373, P〈0.05） ;that of IBS-2 group was 0.53 ± 0.01, and compared with that of IBS-3 group, the difference was statistically significant （t =-3. 139, P〈0. 05）. The MTT value of CD8^＋ T ceils proliferation stimulated by DC was 0. 52±0.01 and 0. 59±0.00（t=3. 539,P〈0.01） ; that of IBS-2 group was 0.54±0.01, and compared with that of IBS-3 group, the difference was statistically significant （t= -3. 183,P〈0.05）. The level of IL-4 secreted by CD4^＋ T cells promoted by DC of bla

关 键 词：PDIA3 树突细胞 白细胞介素4 白细胞介素9 CD4阳性T淋巴细胞 CD8阳性T淋巴细胞 内脏高敏感 

分 类 号：R574.4[医药卫生—消化系统]