DataSheet1_Protective Effect of Prim-O-Glucosylcimifugin on Ulcerative Colitis and Its Mechanism.doc

Intestinal epithelial immune dysfunction or imbalance in the homeostasis of intestinal flora can lead to the occurrence or exacerbation of ulcerative colitis (UC). Prim-O-glucosylcimifugin (POG) is an extract of Chinese traditional medicine (TCM) Saposhnikov, which has analgesic, anti-inflammatory, and antioxidant effects. The present work discussed how the POG alternated ulcerative colitis (UC) along with its underlying mechanism. This was clarified by performing animal studies in a mice model, wherein UC was induced by dextran sulfate sodium (DSS). In vivo studies have found that POG increased clinical score, colonic length, and weight of mice in the ulcerative colitis model. It repaired the pathological injury of an intestinal mucosa within mice while inhibiting the inflammatory factor levels such as IL-1β, TNF-α, and IL-6. Meanwhile, by16SrDNA sequencing analysis, it was found that POG regulated the richness of intestinal microbiota structure and repaired the intestinal immune barrier by upregulating the expression levels of tight junction proteins Occludin, Claudin-3, and ZO-1. To further confirm the above results, we found in in vitro studies that POG also protected lipopolysaccharide- (LPS-) induced RAW264.7 cells. POG dramatically suppressed inflammatory factor production (including TNF-α, IL-1β, and IL-6) within LPS-treated RAW264.7 cells by inhibiting the activation of ERK1/2, AKT, JNK1/2, IκB-α, P38, and P65 phosphorylation. In conclusion, POG plays a protective role against UC by inhibiting the activation of pro-inflammatory signaling pathways MAPK, AKT, and NF-κB; repairing the integrity of the intestinal barrier; and regulating the diversity and abundance of intestinal flora.

肠上皮免疫功能异常或肠道菌群稳态失衡可诱发或加重溃疡性结肠炎（ulcerative colitis, UC）。前定-O-葡萄糖基升麻素（Prim-O-glucosylcimifugin, POG）是传统中医药（Chinese traditional medicine, TCM）防风属植物的提取物，其具备镇痛、抗炎及抗氧化活性。本研究探讨了POG对溃疡性结肠炎的干预作用及其潜在分子机制。本研究通过硫酸葡聚糖钠（dextran sulfate sodium, DSS）诱导构建小鼠溃疡性结肠炎模型开展体内实验。体内研究结果显示，POG可提升溃疡性结肠炎模型小鼠的临床评分、结肠长度及体质量；修复小鼠肠黏膜病理损伤，同时抑制IL-1β、TNF-α及IL-6等炎性因子的表达水平。与此同时，通过16S rDNA测序分析发现，POG可调节肠道菌群结构的丰富度，并通过上调紧密连接蛋白Occludin、Claudin-3及ZO-1的表达水平修复肠道免疫屏障。为进一步验证上述结论，本研究开展了体外实验，结果显示POG同样可保护脂多糖（lipopolysaccharide, LPS）诱导的RAW264.7细胞。POG可通过抑制ERK1/2、AKT、JNK1/2、IκB-α、P38及P65的磷酸化激活，显著下调经LPS处理的RAW264.7细胞中TNF-α、IL-1β及IL-6等炎性因子的产生。综上，POG可通过抑制促炎信号通路MAPK、AKT及NF-κB的激活、修复肠屏障完整性以及调节肠道菌群的多样性与丰度，对溃疡性结肠炎发挥保护作用。