Qingchang Wenzhong Decoction ameliorates intestinal inflammation and intestinal barrier dysfunction in ulcerative colitis via the GC-C signaling pathway
Qingchang Wenzhong Decoction ameliorates intestinal inflammation and intestinal barrier dysfunction in ulcerative colitis via the GC-C signaling pathway
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DOI：
10.1016/j.jep.2023.117503
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发表时间：
2023-12-17
影响因子：
5.4
通讯作者：
Shi，Lei
中科院分区：
文献类型：
--
作者：
Zhang，Yang;Li，Junxiang;Shi，Lei
溃疡性结肠炎（UC）是一种结肠粘膜的特发性慢性炎症性疾病，伴有腹痛和血性腹泻。目前，UC的临床治疗选择有限。清肠温中汤是治疗UC的有效中药方剂。然而，QCWZD在缓解UC肠屏障功能障碍的机制还没有清楚地explained.Aim的studyTo确定的机制，QCWZD促进肠屏障功能障碍的恢复在UC. MaterialsandMethodsA二次分析从UC患者获得从以前的RCT临床试验的结肠粘膜进行。使用UC患者的结肠组织石蜡包埋切片评价QCWZD对UC患者肠粘液和机械屏障的影响。并通过体内、体外实验进一步探讨其作用机制。UC小鼠在含有3.0%葡聚糖硫酸钠（DSS）的无菌水中建立。同时，治疗组小鼠给予QCWZD或美沙拉嗪。在体外，使用Caco-2和HT 29细胞共培养构建肠屏障模型。GC-C质粒用于过表达/敲低GC-C以阐明QCWZD的靶标。采用HE、AB-PAS、ELISA、免疫组织化学和免疫荧光法检测结肠炎症程度和肠屏障完整性。RT-qPCR、Western Blot检测GC-C信号通路相关基因和蛋白的表达。结果在UC患者中，QCWZD增加粘液分泌，杯状细胞数量，并促进MUC 2和ZO-1的表达。QCWZD加速UC小鼠从DSS诱导的炎症中恢复，包括体重增加、疾病活动指数（DAI）评分降低、结肠长度恢复和组织学愈合。在体内和体外实验中，QCWZD促进粘液分泌并增加ZO-1表达，从而修复粘液机械屏障损伤。QCWZD的作用通过调节GC-C信号通路介导，进而影响CFTR磷酸化和MUC 2表达以促进粘液分泌，同时抑制MLCK的过度激活并修复紧密连接以维持机械屏障的完整性。分子对接的结果表明，结合的主要成分QCWZD GC-C。结论我们的研究表明，QCWZD调节GC-C信号通路，以促进缓解粘液机械屏障损伤的UC。QCWZD作用机制的阐明为UC新疗法的开发提供了希望。
Ethnopharmacological relevanceUlcerative colitis (UC) is an idiopathic, chronic inflammatory disorder of the colonic mucosa, accompanied with abdominal pain, and bloody diarrhea. Currently, clinical treatment options for UC are limited. Qingchang Wenzhong Decoction (QCWZD) is an effective prescription of traditional Chinese medicine for the treatment of UC. However, the mechanism of QCWZD in alleviating intestinal barrier dysfunction in UC has not been clearly explained.Aim of the studyTo determine the mechanism whereby QCWZD promotes the recovery of intestinal barrier dysfunction in UC.Materials and methodsA secondary analysis of colonic mucosa from UC patients acquired from a prior RCT clinical trial was performed. The effects of QCWZD on intestinal mucus and mechanical barriers in UC patients were evaluated using colon tissue paraffin-embedded sections from UC patients. The mechanism was further investigated by in vivo and in vitro experiments. UC mice were established in sterile water with 3.0% dextran sodium sulfate (DSS). Meanwhile, mice in the treatment group were dosed with QCWZD or mesalazine. In vitro, an intestinal barrier model was constructed using Caco-2 and HT29 cells in co-culture. GC-C plasmid was used to overexpress/knock down GC-C to clarify the target of QCWZD. HE, AB-PAS, ELISA, immunohistochemistry and immunofluorescence assays were used to assess the level of colonic inflammation and intestinal barrier integrity. Rt-qPCR, Western Blot were used to detect the expression of genes and proteins related to GC-C signaling pathway. Molecular docking was used to simulate the binding sites of major components of QCWZD to GC-C.ResultsIn UC patients, QCWZD increased mucus secretion, goblet cell number, and promoted MUC2 and ZO-1 expression. QCWZD accelerated the recovery of UC mice from DSS-induced inflammation, including weight gain, reduced disease activity index (DAI) scores, colon length recovery, and histological healing. QCWZD promoted mucus secretion and increased ZO-1 expression in in vivo and in vitro experiments, thereby repairing mucus mechanical barrier damage. The effects of QCWZD are mediated through regulation of the GC-C signaling pathway, which in turn affects CFTR phosphorylation and MUC2 expression to promote mucus secretion, while inhibiting the over-activation of MLCK and repairing tight junctions to maintain the integrity of the mechanical barrier. Molecular docking results demonstrate the binding of the main components of QCWZD to GC-C.ConclusionOur study demonstrated that QCWZD modulates the GC-C signaling pathway to promote remission of mucus-mechanical barrier damage in the UC. The clarification of the mechanism of QCWZD holds promise for the development of new therapies for UC.