摘要
目的探究IL-34基因敲除对小鼠肠黏膜通透性的影响及相关机制。方法选取20只6周龄SPF级雄性C57BL/6野生型(WT)小鼠作为研究对象,采用随机数表法分为WT组(WT小鼠)和KO组(IL-34^(-/-)小鼠),每组10只。通过连续称量记录14周两组的体质量,观察IL-34基因敲除对小鼠体质量的影响。采用H-E染色法观察两组小鼠结肠的病理改变。采用免疫组织化学染色法、实时荧光定量PCR(real-time qPCR)法检测两组小鼠结肠组织中紧密连接蛋白ZO-1和Occludin的表达。利用异硫氰酸荧光素标记的葡聚糖(FITC-dextran)评估IL-34基因敲除后对小鼠肠黏膜通透性的影响。在Caco-2细胞(人结直肠腺癌细胞)中观察IL-34对ZO-1和Occludin表达的影响。结果KO组与WT组在14周内体质量的变化情况差异无统计学意义。H-E染色结果显示,WT组和KO组的结肠均无明显损伤表现。免疫组织化学染色结果显示,与WT组比较,KO组结肠组织中ZO-1和Occludin的表达水平均较低(P均<0.05)。KO组的血清FITC-dextran水平显著高于WT组(P<0.05)。与WT组比较,KO组的结肠组织中ZO-1 mRNA和Occludin mRNA的相对表达量均较低(P均<0.05)。蛋白质印迹法结果显示,IL-34可上调Caco-2细胞中ZO-1和Occludin的表达。结论IL-34基因敲除可增高结肠黏膜的通透性,这可能是通过调控结肠组织中ZO-1和Occludin的表达实现的。
Objective This paper is to study the effects of IL-34 gene knockout on intestinal mucosal permeability in mice.Methods Twenty 6-week-old SPF-grade male C57BL/6 wild-type(WT)mice were selected as the research objects,and divided into WT group(WT mice)and KO group(IL-34^(-/-)mice)by random number table method,10 in each group.The body weights of the two groups were recorded by continuous weighing for 14 weeks,and the effect of IL-34 gene knockout on the body weight of mice was observed.The colonic pathological changes of the two groups of mice were observed by H-E staining.The expressions of tight-junction proteins ZO-1 and Occludin in the colon tissue of the two groups of mice were detected by immunohistochemical staining and real-time qPCR.The effects of IL-34 gene knockout on intestinal mucosal permeability in mice were evaluated by using the FITC-dextran method.The effects of IL-34 on ZO-1 and Occludin were observed in Caco-2 cells(human colorectal adenocarcinoma cells).Results There is no significant difference in body weight changes between the KO group and the WT group within 14 weeks.The results of H-E staining shows that there is no obvious damage to the colon of the WT group and the KO group.The results of immunohistochemical staining shows that compared with the WT group,the expressions of ZO-1 and Occludin in the colon tissue of the KO group are lower(P<0.05).The serum FITC-dextran level of the KO group is significantly higher than that of the WT group(P<0.05).Compared with the WT group,the relative expressions of ZO-1 mRNA and Occludin mRNA in the colon tissue of the KO group are lower(P<0.05).The results of western blotting shows that IL-34 can up-regulate the expressions of ZO-1 and Occludin in Caco-2 cells.Conclusions The knockout of IL-34 gene can increase the permeability of the colonic mucosa.It may be obtained by upregulating the expression of ZO-1 and Occludin proteins in the colon tissues.
作者
丁琴
李文文
刘渊
DING Qin;LI Wenwen;LIU Yuan(Department of Gastroenterology,the Sixth People's Hospital Affiliated to Shanghai Jiao Tong University,Shanghai 200233,China)
出处
《国际消化病杂志》
CAS
2022年第2期120-125,共6页
International Journal of Digestive Diseases
基金
国家自然科学基金资助项目(81470833)。
