白介素35在炎性肠疾病中的抗炎作用机制  

Anti-inflammatory effect of interleukin-35 in mice with colitis and its mechanism

作　　者：卢战军 胡洋洋[2,3] 李思思 臧丽娟 蒋巍亮[2,3] 吴坚炯 巫协宁[2] 曾悦[2,3] 王兴鹏[1,2,3] LU Zhanjun;HU Yangyang;LI Sisi;ZANG Lijuan4;JIANG Weiliang;WU Jianjiong;WU Xiening;ZENG Yue;WANG Xingpeng(Department of Gastroenterology,Shanghai General Hospital of Nanjing Medical University,Shanghai 200080,China;Department of Gastroenterology,Shanghai General Hospital,Shanghai 200080,China;Shanghai Key Laboratory of Pancreatic Disease,Shanghai 200080,China;Pathology Center,Shanghai General Hospital,Shanghai 200080,China)

机构地区：[1]南京医科大学附属上海一院临床医学院消化科,上海200080 [2]上海交通大学附属第一人民医院消化科,上海200080 [3]上海市胰腺疾病重点实验室,上海200080 [4]上海交通大学附属第一人民医院病理中心,上海200080

出　　处：《浙江大学学报（医学版）》2018年第5期499-506,共8页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金(81802318)

摘　　要：目的:研究IL-35在炎性肠疾病中的抗炎作用及相关机制。方法:BALB/c雌性小鼠共30只,随机分为对照组、模型组(口服4%葡聚糖硫酸钠7 d)、IL-35组(口服4%葡聚糖硫酸钠7 d,第2～5天腹腔注射IL-35 2μg/d),每组10只。每天对小鼠进行疾病活动指数(DAI)评分; 7 d后处死小鼠,留取血清和肠道组织,观察结肠大体形态; HE染色观察各组结肠组织病理形态变化;流式细胞术检测各组结肠组织中巨噬细胞极化情况;实时定量RT-PCR检测各组结肠组织中细胞因子IL-6、TNF-α、γ干扰素(IFN-γ)、IL-10和Src同源系列2结构域的肌醇5-磷酸酶1(SHIP1)的mRNA表达量;免疫组织化学法检测各组结肠组织中SHIP1的表达及分布情况;蛋白质印迹法检测各组结肠组织中SHIP1蛋白的表达。结果:模型组在实验过程中DAI评分较对照组增加,而IL-35组自第4天起DAI评分较模型组减少(均P<0.01);与对照组比较,模型组结肠明显缩短(P<0.05),而IL-35组的结肠长度长于模型组,但差异无统计学意义(P>0.05);与模型组比较,IL-35组炎症细胞浸润减少、黏膜组织炎症评分和腺窝破坏组织评分较模型组减少(均P<0.05); IL-35组结肠组织中促炎因子IL-6、TNF-α和IFN-γ的mRNA相对表达量较模型组减少,而抑炎因子IL-10的mRNA相对表达量较模型组增加(均P<0.05);与对照组比较,模型组M1型巨噬细胞比例增加(P<0.05),而IL-35组M1型巨噬细胞的比例较模型组减少(P<0.05); IL-35组小鼠结肠组织中SHIP1mRNA和蛋白相对表达量均较模型组增加(均P<0.05)。结论:在炎性肠疾病中,IL-35可以通过调控SHIP1表达抑制M1型巨噬细胞极化,以及调节炎症因子的表达发挥抗炎作用。Objective:To investigate the anti-inflammatory effect and mechanisms of interleukin-35(IL-35) in inflammatory bowel disease.Methods:BALB/c mice were divided into three groups with 10 mice in each group:control group,model group(oral administration of 4% glucan sodium sulfate for 7 d) and IL-35-treated group(oral administration of 4% glucan sodium sulfate for 7 d,intraperitoneal injection of 2 μg IL-35 at d2-5).Disease activity index(DAI) was scored every day.After 7 d,the mice were sacrificed,and the serum and intestinal tissue samples were collected.The gross morphology of the colon was observed;HE staining was used to observe the pathological changes of colon tissue;flow cytometry was employed to detect the change of macrophage polarization ratio in colon tissue;the mRNA expression levels of cytokines IL-6,TNF-α,IFN-γ,IL-10 and SHIP1 in colon tissue were determined by real-time quantitative RT-PCR;the expression and distribution of SHIP1 in colon tissue was measured by immunohistochemistry;Western blotting was adopted to detect the expression level of SHIP1 protein in colonic intestinal tissues of each group.Results:The DAI scores of the mice in the model group were higher than those in the control group,while the DAI scores in the IL-35-treated group were lower than those in the model group(all P<0.01).Compared with the control group,the colon length was significantly shortened in the model group(P<0.05),while the colon length of the IL-35-treated group had an increasing trend compared with the model group,but the difference was not statistically significant(P>0.05).Compared with the model group,microscopic inflammatory infiltration score was decreased and microscopic crypt destruction and score was significantly lower in IL-35-treated group(all P<0.05).The relative expression of proinflammatory cytokines IL-6,TNF-α and IFN-γ in the colon tissue of IL-35-treated group was decreased compared with the model group,while the relative expression of IL-10 mRNA was higher than that of the model group(all P<0.05)

关 键 词：炎性肠疾病/病理生理学 白细胞介素类/代谢 免疫组织化学 印迹法 蛋白质 随机对照试验 疾病模型 动物 

分 类 号：R574[医药卫生—消化系统]