摘要
目的分析肿瘤坏死因子样配体1A(tumor necrosis factor ligand-related molecule1A,TL1A)表达对炎症性肠病相关肠纤维化的意义。方法C57BL/6野生型小鼠,9~12周,平均(10.61±0.17)周;体重20~24 g,平均(22.50±0.19)g;清洁级,淋系细胞高表达TL1A转基因型小鼠,8~12周,平均(10.57±0.20)周;体重19~25 g,平均(22.61±0.23)g;清洁级,以Masson(MT)染色与天狼猩红染色法定量分析肠组织纤维化程度。以免疫组织化学法对结肠组织内Ⅰ、Ⅲ型胶原、TGF-β1及Smad3表达进行测定。以右旋葡聚糖硫酸钠诱导建立炎症性肠病相关肠纤维化小鼠模型,采取随机法将C56BL/6野生小鼠及基因型小鼠分作研究组与对照组,每组10只。对照组予以饮用蒸馏水,研究组予以间断性饮用2%右旋葡聚糖硫酸钠水,共4周。研究组右旋葡聚糖硫酸钠水饮用时间为:d1~5,d8~12,d15~19,d22~26,其余时间均饮用蒸馏水。定期观察小鼠,评定其结肠纤维化程度,结肠组织Ⅰ、Ⅲ型胶原表达,转化生长因子-β1(transforming growth factor-β1,TGF-β1)及Smad3表达。结果以LCK-CD2-TL1A-GFP引物序列对小鼠DNA进行测定,于192 bp处,野生小鼠无目的基因检出,而转基因小鼠则可见目的基因成像,以此可鉴定出转基因小鼠。健康结肠黏膜组织中有少量胶原蛋白沉积,予以MT染色后,呈现出轻微蓝色;予以天狼猩红染色后,呈现出红色,且于纤维化进程中,黏膜组织及黏膜组织下层胶原蛋白的沉积量逐渐增加,染色程度、染色面积也随之增加。野生小鼠中,研究组结肠纤维化程度加重比对照组更高;转基因小鼠中,研究组结肠纤维化程度加重比对照组更高,且转基因小鼠结肠纤维化程度比野生小鼠高,差异有统计学意义(P<0.05);野生小鼠中,研究组小鼠Ⅰ、Ⅲ型胶原表达均比对照组高;转基因小鼠中,研究组小鼠的Ⅰ、Ⅲ型胶原表达比对照组更高,且转基因小鼠Ⅰ、Ⅲ型胶原表达比野生小鼠高,差异有统计学意义(P<0.05);野生小鼠中,研究组小鼠的TGF-β1及Smad3表达均比对照组高;转基因小鼠中,研究组小鼠的TGF-β1及Smad3表达比对照组更高,且转基因小鼠结肠组织中TGF-β1及Smad3表达比野生小鼠高,差异有统计学意义(P<0.05)。结论TL1A对肠成纤维细胞组织活化及增殖起促进作用,通过促使Ⅰ、Ⅲ型胶原大量合成,使炎症性肠病相关肠纤维化的程度加重,同时TL1A还能使炎症性肠病相关肠纤维化进一步发展,TL1A表达对炎症性肠病相关肠纤维化的作用机制之一即促进TGF-β1及Smad3表达上调。
Objective To investigate the significance of tumor necrosis factor ligand-related molecule1A(TL1A)expression in intestinal fibrosis related with inflammatory bowel diseases.Methods The mouse models with intestinal fibrosis related with inflammatory bowel diseases were established by induction with dextran sodium dextran sulfate,and the C56BL/6 wild mice and genotype mice were divided into research group and control group,respectively,with 10 mice in each group.The mice in control group were given distilled water,however,the mice in research group were given intermittently drinking 2%sodium dextran sulfate sodium sulfate,and the drinking time of dextran sulfate sodium sulfate in research group was d1~5,d8~12,d15~19,d22~26,with dranking distilled water for the rest of time,with a course of 4 weeks for both groups.The degree of colonic fibrosis,the expression levels of typeⅠandⅢcollagen in colon tissue,transforming growth factor-β1(TGF-β1)and Smad3 were observed and compared between the two groups.Results The degree of colonic fibrosis in research group was significantly severer than that in control group.The expression levels of typeⅠandⅢcollagen in research group were significantly higher than those in control group.Moreover the expression levels of TGF-β1 and Smad3 in research group were significantly higher than those in control group,which in transgenic mice were significantly higher than those in mild mice(P<0.05).Conclusion TL1A can promote the activation and proliferation of intestinal fibroblasts by promoting the mass synthesis of type I and III collagen,increasing the degree of intestinal fibrosis related with inflammatory bowel diseases,which can also promote the further development of the intestinal,and one of the action mechanisms of TL1A is to up-regulate the expressions of TGF-β1 and Smad3.
作者
杨玉宇
阮巍山
徐莉
陈东锋
YANG Yuyu;RUAN Weishan;XU Li(Department of Gastroenterology,Zhongshan People’s Hospital,Guangdong,Zhongshan 528403,China)
出处
《河北医药》
CAS
2021年第11期1605-1610,共6页
Hebei Medical Journal
基金
广东省科技发展专项资金(公益研究与能力建设方向)(编号:2017B030314037)。
关键词
肿瘤坏死因子样配体1A
炎症性肠病
肠纤维化
转化生长因子-Β1
SMAD3表达
tumor necrosis factor ligand-related molecule1A
inflammatory bowel disease
bowel fibrosis
transforming growth factor-β1
Smad3 expression
