巨噬细胞表型改变在先天性巨结肠相关性小肠结肠炎中的研究  

Phenotypic changes of macrophages in Hirschsprung's disease associated enterocolitis

作　　者：陈绪勇[1] 焦春雷[1] 付康[1] 冯晨钊 朱天琦[1] 柯昌庶[3] 冯杰雄[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院小儿外科,武汉430030 [2]武汉外国语学校 [3]华中科技大学同济医学院附属同济医院病理科,武汉430030

出　　处：《腹部外科》2016年第6期467-472,共6页Journal of Abdominal Surgery

基　　金：国家自然科学基金(81270441)

摘　　要：目的通过检测先天性巨结肠患儿结肠组织中巨噬细胞表型的改变,探讨巨噬细胞活化与巨结肠相关性肠炎发病之间的关系。方法收集先天性巨结肠患儿(肠炎组4例,非肠炎组6例)结肠远段及近段标本,行HE染色、评估各段肠管炎症损伤程度,CD68和iNOS免疫荧光双染法评估M1型巨噬细胞活化状况,CD68和Arg-1免疫荧光双染法评估M2型巨噬细胞活化状况,RTqPCR检测各组肠管iNOS及Arg-1的mRNA表达水平。结果免疫荧光染色显示,肠炎组近段结肠iNOS/C/D68阳性细胞的百分比(73.12±2.48)%明显高于肠炎组远段结肠(54.19±1.98)%、非肠炎组近段结肠(49.35±2.70)%及远段结肠(43.18±4.21)%;而肠炎组近段结肠Arg-1/CD68阳性细胞的百分比(33.15±4.81)%明显低于远段结肠(49.32±3.98)%;非肠炎组近段结肠Arg-1/CD68阳性细胞的百分比(45.23±1.95)%亦低于远段结肠(56.52±2.79)%,差异有统计学意义(P<0.05)。RT-qPCR结果显示,肠炎组近段结肠iNOS表达明显升高,而Arg-1表达则在肠炎组及非肠炎组的远段结肠升高(P<0.05)。结论先天性巨结肠相关性小肠结肠炎的发生可能与巨噬细胞向M1型转化有关。Objective To investigate the phenotypic changes of macrophages in patients with Hirschsprung disease, and to explore the role of macrophage activation in Hirschsprung＇ s associated entercolitis. Methods The distal segment of colon and proximal segments of the colon specimens in 10 patients with Hirschsprung＇s disease （Entercolitis group： 4 cases; non-entercolitis group： 6 cases） were collected. HE staining was used to assess the degree of bowel inflammation injury. CD68 and iNOS immunofluorescence double staining was used to assess M1 macrophages, and CD68 and Arg-1 immunofluorescence double staining was used to assess M2 macrophages. The mRNA expression levels of iNOS and Arg-1 were detected by RT-qPCR. Results Immunofluorescence staining showed that the percentage of iNOS/CD68 positive cells in proximal segments of HAEC group [（73. 12± 2. 48）%] was higher than that of distal segments of HAEC group [（54. 19±1.98）%], proximal segments of HD group [（49.35 ± 2. 70） %] and distal segments of HD group [（43. 18± 4. 21 ） %]. But the percentage of Arg-1/CD68 positive cells in proximal segment colon of HAEC group [（33. 15 ＋ 4. 81）%] was lower than in distal segment of HAEC group [（49. 32 ± 3. 98） %], and percentage of Arg-1/CD68 positive cells in proximal segment colon of HD group [（45.23 ± 1.95）%] was also lower than in distal segment of HD group [（56. 52± 2. 79）%] （PM. 05）. RT-qPCR results showed that the expression of iNOS was significantly increased in proximal segments of colon in HAEC group, but that of Arg-1 was higher in distal segment of HAEC group （P〈0. 05）. Conclusions The occurrence of Hirschsprung＇s associated entercolitis may be associated with the activation of M1 macrophages.

关 键 词：先天性巨结肠相关性小肠结肠炎 巨噬细胞表型 

分 类 号：R656.9[医药卫生—外科学]