ABSTRACT
OBJECTIVE: Intestinal fibrosis is a refractory complication of inflammatory bowel disease (IBD). Tumor necrosis factor ligand-related molecule-1A (TL1A) is important for IBD-related intestinal fibrosis in a dextran sodium sulfate (DSS)-induced experimental colitis model. This study aimed to explore the effects of TL1A on human colonic fibroblasts. METHODS: A trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis model of LCK-CD2-TL1A-GFP transgenic (Tg) or wild-type (WT) mice was established to determine the effect and mechanism of TL1A on intestinal fibrosis. The human colonic fibroblast CCD-18Co cell line was treated concurrently with TL1A and human peripheral blood mononuclear cell (PBMC) supernatant. The proliferation and activation of CCD-18Co cells were detected by BrdU assays, flow cytometry, immunocytochemistry and Western blotting. Collagen metabolism was tested by Western blotting and real-time quantitative polymerase chain reaction (RT-qPCR). RESULTS: The level of collagen metabolism in the TNBS+ethyl alcohol (EtOH)/Tg group was greater than that in the TNBS+EtOH/WT group. Transforming growth factor-ß1 (TGF-ß1) and p-Smad3 in the TNBS+EtOH/Tg group were upregulated as compared with those in the TNBS+EtOH/WT group. The proliferation of CCD-18Co cells was promoted by the addition of human PBMC supernatant supplemented with 20 ng/mL TL1A, and the addition of human PBMC supernatant and TL1A increased CCD-18Co proliferation by 24.4% at 24 h. TL1A promoted cell activation and increased the levels of COL1A2, COL3A1, and TIMP-1 in CCD-18Co cells. Treatment of CCD-18Co cells with TL1A increased the expression of TGF-ß1 and p-Smad3. CONCLUSION: TL1A promotes TGF-ß1-mediated intestinal fibroblast activation, proliferation, and collagen deposition and is likely related to an increase in the TGF-ß1/Smad3 signaling pathway.
Subject(s)
Cell Proliferation , Fibroblasts , Fibrosis , Signal Transduction , Smad3 Protein , Transforming Growth Factor beta1 , Tumor Necrosis Factor Ligand Superfamily Member 15 , Tumor Necrosis Factor Ligand Superfamily Member 15/metabolism , Tumor Necrosis Factor Ligand Superfamily Member 15/genetics , Smad3 Protein/metabolism , Smad3 Protein/genetics , Humans , Fibroblasts/metabolism , Fibroblasts/pathology , Animals , Transforming Growth Factor beta1/metabolism , Transforming Growth Factor beta1/genetics , Mice , Colon/metabolism , Colon/pathology , Colitis/metabolism , Colitis/chemically induced , Colitis/pathology , Colitis/genetics , Cell Line , Mice, Transgenic , Trinitrobenzenesulfonic Acid , Disease Models, Animal , Leukocytes, Mononuclear/metabolismABSTRACT
OBJECTIVE: By sequenceing the Cj1136, Cj1138 and Cj1139 gene of Campylobacter jejuni (C. jejuni) strains associated with Guillain-Barré Syndrome (GBS), features of Cj1136, Cj1138 and Cj1139 gene were studied. Results were compared with the C. jejuni strain NCTC11168, to find the mutations in sequence of C. jejuni which inducing GBS and their polygenetic relationship was analyzed. METHODS: Three GBS-associated C. jejuni strains were isolated from stools of GBS patients from Hebei province who had been diagnosed as clinical AMAN pattern and electrophysiological tests were performed. After distilling and sequencing Cj1136, Cj1138 and Cj1139 genes, results were spliced and assembled into a complete sequence by the terminals overlapped with each other. Sequences of Cj1136, Cj1138 and Cj1139 genes were compared with NCTC11168, to find the mutations and gene feature. RESULTS: The Cj1136, Cj1138 and Cj1139 gene of the three GBS- associated C. jejuni strains were composed by 1173 base pairs, 1170 base pairs, 912 base pairs respectively. The alignment with the related sequence of NCTC11168 showed that there were two same mutations in the Cj1138 gene of the three C. jejuni stains. Data from phylogenetic analysis demonstrated that the three C. jejuni strains were genetically closed to NCTC11168, with the biggest phylogenetic distance between the three of them as 2.1%. CONCLUSION: When compared with NCTC11168 the Cj1138 gene of the three GBS-associated C. jejuni strains had the same mutations which might be related to the development of GBS. Relation between the variation and GBS-pathogenesis remained to be confirmed. The mutations found in the three C. jejuni strains established the foundation for exploring the biological characteristics of GBS-associated C. jejuni strains and demonstrated that the GBSassociated C. jejuni strains of Hebei province having its regional features.
