LAMB3 Promotes Intestinal Inflammation Through SERPINA3 and Is Directly Transcriptionally Regulated by P65 in Inflammatory Bowel Disease.
LAMB3 Promotes Intestinal Inflammation Through SERPINA3 and Is Directly Transcriptionally Regulated by P65 in Inflammatory Bowel Disease.
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LAMB3通过SERPINA3促进肠道炎症，并在炎症性肠病中由P65直接转录调节。
DOI：
10.2139/ssrn.4139688
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发表时间：
2023-07
影响因子：
4.9
通讯作者：
Fangyuan Liu;Weimin Xu;Yaosheng Wang;Zhenyu Huang;Zhehui Zhu;Weijun Ou;Wenbo Tang;Jihong Fu;Chenying Liu;Yubei Gu;Yun Liu;Peng Du
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文献类型：
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作者：
Fangyuan Liu;Weimin Xu;Yaosheng Wang;Zhenyu Huang;Zhehui Zhu;Weijun Ou;Wenbo Tang;Jihong Fu;Chenying Liu;Yubei Gu;Yun Liu;Peng Du
各种细胞外基质（ECM）重塑事件参与炎症性肠病（IBD）。LAMB 3是层粘连蛋白-332的重要亚基，层粘连蛋白-332是一种重要的ECM组分。LAMB 3在肠道炎症中的生物学功能的数据缺乏。我们的目的是讨论LAMB 3在IBD中的作用。方法在培养的肠上皮细胞、患者炎症粘膜组织和小鼠结肠炎模型中评估LAMB 3的表达。采用RNA测序、实时荧光定量PCR和Western blotting检测LAMB 3的表达分布和潜在的下游靶基因。采用双荧光素酶法和染色质免疫沉淀-定量聚合酶链反应检测肿瘤坏死因子α刺激下P65是否能转录激活LAMB 3。结果LAMB 3表达在肠上皮细胞和结肠样组织的炎症状态中增加，并与克罗恩病的不良临床结局相关。LAMB 3的敲低抑制促炎细胞因子的表达。在机制上，LAMB 3的表达被P65直接转录激活，并在肿瘤坏死因子α刺激下被核因子κ B抑制剂抑制。此外，RNA测序和补充实验表明，LAMB 3上调SERPINA 3，通过整合素α3β1/FAK途径促进肠道炎症。结论LAMB 3可能成为IBD治疗的一个潜在靶点，并可作为克罗恩病肠狭窄的一个预测因子。我们的研究结果表明，ECM在IBD的发展中起着重要作用，并为IBD的治疗和预后提供了实验依据。
BACKGROUND Various extracellular matrix (ECM) reshaping events are involved in inflammatory bowel disease (IBD). LAMB3 is a vital subunit of laminin-332, an important ECM component. Data on the biological function of LAMB3 in intestinal inflammation are lacking. Our aim is to discuss the effect of LAMB3 in IBD. METHODS LAMB3 expression was assessed in cultured intestinal epithelial cells, inflamed mucosal tissues of patients and mouse colitis models. RNA sequencing, quantitative real-time polymerase chain reaction and Western blotting were used to detect the LAMB3 expression distribution and potential downstream target genes. Dual-luciferase assays and chromatin immunoprecipitation-quantitative polymerase chain reaction were used to determine whether P65 could transcriptionally activate LAMB3 under tumor necrosis factor α stimulation. RESULTS LAMB3 expression was increased in inflammatory states in intestinal epithelial cells and colonoids and was associated with adverse clinical outcomes in Crohn's disease. Knockdown of LAMB3 inhibited the expression of proinflammatory cytokines. Mechanistically, LAMB3 expression was directly transcriptionally activated by P65 and was inhibited by nuclear factor kappa B inhibitors under tumor necrosis factor α stimulation. Furthermore, RNA sequencing and replenishment experiments revealed that LAMB3 upregulated SERPINA3 to promote intestinal inflammation via the integrin α3β1/FAK pathway. CONCLUSION We propose that LAMB3 could serve as a potential therapeutic target of IBD and a predictor of intestinal stenosis of Crohn's disease. Our findings demonstrate the important role of ECM in the progression of IBD and offer an experimental basis for the treatment and prognosis of IBD.