半乳糖凝集素-受体相互作用对旋毛虫感染小鼠肠道黏膜免疫的调节

The galectin-receptor interaction may regulate intestinal mucosal immunity of mice infected with Trichinella spiralis

目的探讨半乳糖凝集素对旋毛虫感染小鼠小肠组织的免疫调节作用。方法本实验共使用26只雌性昆明小鼠,其中正常对照组6只;α-乳糖对照组6只,每天给每只鼠经腹腔注射200μL 1.5 mmol/Lα-乳糖溶液;旋毛虫感染组7只,每只鼠口饲感染旋毛虫幼虫300条;旋毛虫感染+α-乳糖组7只,每只鼠口饲感染旋毛虫幼虫300条,小鼠感染后115 d每天每只鼠经腹腔注射200μL 1.5 mmol/Lα-乳糖溶液。感染旋毛虫后第15天处死小鼠,取小肠,采用HE染色观察小肠组织病理变化,采用Image-Pro Plus 6.0软件对小肠组织中的杯状细胞和潘氏细胞进行计数,实时荧光定量PCR检测小肠组织中肠道黏蛋白2(MUC2)的相对表达量。采用SPSS 25.0对数据进行多组间单因素方差分析,采用LSD-t检验进行组间两两比较,采用GraphPad Prism 7.0绘制统计图。结果在感染后第15天,与正常对照组和α-乳糖组相比,旋毛虫感染组、旋毛虫感染+α-乳糖组小鼠的小肠组织均出现较明显的病理变化和炎症细胞浸润,杯状细胞和潘氏细胞的数量显著增加,差异有统计学意义(P<0.05),小肠组织中MUC2的mRNA表达水平显著增加,差异有统计学意义(P<0.05)。与旋毛虫感染组相比,旋毛虫感染+α-乳糖组小鼠小肠组织中杯状细胞和潘氏细胞的数量显著增加,差异有统计学意义(P<0.05),小肠组织中MUC2的mRNA表达水平显著增加,差异有统计学意义(P<0.05)。结论使用α-乳糖处理后,旋毛虫感染小鼠小肠组织中杯状细胞和潘氏细胞的数量以及MUC2的mRNA表达水平均增加,提示阻断半乳糖凝集素-受体的相互作用可增强小鼠抗旋毛虫感染的肠道黏膜免疫。

Objective To investigate the effect of galectins-receptor interactions on immune regulation of small intestine of mice infected with Trichinella spiralis.Methods A total of 26 female Kunming mice were used in this experiment.Mice were randomized into 4 groups,(i)uninfected control group,6 mice;(ii)α-lactose-treated group,6 mice,each was injected with 200μL 1.5 mmol/Lα-lactose everyday;(iii)T.spiralis-infected group,7 mice,each was infected with 300 T.spiralis larvae by oral gavage,and(iv)T.spiralis+α-lactose group,7 mice,each was infected with 300 T.spiralis larvae plusα-lactose treatment.Mice were injected intraperitoneally with 200μL 1.5 mmol/L ofα-lactose solution in phosphate buffer saline(PBS)once daily from 1 to 15 days post infection(p.i.).All mice were sacrificed at day 15 p.i.and their small intestines were taken for further analysis.After HE staining,the pathology of intestinal tissues of mice was observed by light microscopy.The numbers of goblet cells and Paneth cells were counted by Image-Pro Plus 6.0 software.The mRNA expression levels of mucin 2(MUC2)were detected by quantitative real-time reverse transcription polymerase chain reaction.Statistical analysis of the data was performed by one-way ANOVA and then followed by LSD-t multiple comparison test by using SPSS 25.0 software for windows.The graphs were made by using GraphPad Prism software(version 7.0).Results On day 15 p.i.,compared with uninfected mice and uninfected mice withα-lactose treatment,there were obvious small intestine pathology,significantly increased numbers of goblet cells and Paneth cells(P<0.05),and significantly increased mRNA expression levels of MUC2 in the small intestines of T.spiralis-infected mice and T.spiralisinfected mice withα-lactose treatment(P<0.05).However,compared with T.spiralis-infected mice,there were significantly increased goblet cells and Paneth cells(P<0.05),and significantly increased mRNA expression level of MUC2 in the small intestine of T.spiralis-infected mice withα-lactose treatment(P<0.05).Conclusion Treatment withα-lactose can increase the numbers of goblet cells and Paneth cells and MUC2 m RNA expression level,suggesting that blockage of the galectin-receptor interaction may enhance murine intestinal mucosal immunity against T.spiralis infection.