背景:林得(LR),枯草草(Sims)Kosterm的干燥根。,是一种传统的中草药,已经使用了数千年来促进气血循环,舒缓肝脏,治疗腹泻和痢疾.先前的研究发现,LR的乙醇提取物通过调节Th17/Treg平衡发挥抗溃疡性结肠炎(UC)的作用。水提物是LR的经典临床应用形式,但LR水提物(LRWE)对UC的作用及其机制尚不清楚。
目的:目的:UC是一种以肠道炎症为特征的胃肠道疾病,粘膜损伤,和纤维化,它是结肠癌的高危因素之一。然而,仍然缺乏效果令人满意的补救措施。本研究旨在探讨LRWE对UC的疗效及可能的作用机制。
方法:使用回流提取法制备LRWE样品。通过给予2.5%DSS水溶液诱导小鼠结肠炎,通过评估疾病活动评分来评估LRWE的治疗效果,结肠长度,和粪便形态。H&E染色,TEM,Masson染色,应用AB-PAS染色观察结肠组织病理学变化。通过转录组学分析结肠组织中差异表达的基因。TUNEL染色检测细胞凋亡。IL-6、IL-1β等炎症因子的表达,以及p-STAT1、p-JAK2、p-STAT3、Bax、并通过免疫荧光和免疫组织化学检测Bcl-2。occludin的表达,Bcl-2,Bax,和JAK/STAT信号通路相关的重要蛋白通过蛋白质印迹(WB)进行定量。
结果:LRWE减轻了体重减轻,结肠缩短,DAI分数,病理变化,结肠炎小鼠结肠组织的超微结构特征。它还抑制促炎细胞因子(如TNF-α,IL-6和IL-1β)和增加的IL-10水平。此外,它通过上调Occludin和Mucin-2的表达来保护肠屏障。机械上,LRWE可能通过降低p-JAK2、p-STAT3、p-STAT1、Bcl2和Bax的蛋白表达来抑制JAK-STAT信号通路的激活,从而减轻炎症反应,抑制细胞凋亡。
结论:LRWE对DSS诱导的UC具有保护作用。这种效应与JAK-STAT信号通路的抑制有关,改善肠道炎症,减少肠上皮细胞凋亡。
BACKGROUND: Linderae Radix (LR), the dried root of Lindera aggregata (Sims) Kosterm., is a traditional Chinese herbal medicine that has been used for thousands of years for promoting Qi circulation, soothing the liver, and treating diarrhea and dysentery. Previous studies have found that ethanol extract of LR plays an anti-ulcerative colitis (UC) role by regulating Th17/ Treg balance. Water extract is the classic clinical application form of LR, but the effect of water extract of LR (LRWE) on UC and its underlying mechanism is still unclear.
OBJECTIVE: Purpose: UC is a gastrointestinal disease characterized by intestinal inflammation, mucosal injury, and fibrosis, and it is one of the high-risk factors for colon cancer. However, there is still a lack of remedies with satisfactory effects. This study aimed to investigate the efficacy and the potential mechanism of LRWE against UC.
METHODS: LRWE samples were prepared using a reflux extraction method. Colitis in mice was induced by administering 2.5 % DSS water solution to evaluate the therapeutic effect of LRWE by assessing disease activity score, colon length, and fecal morphology. H&E staining, TEM, Masson staining, and AB-PAS staining were applied to observe histopathological changes in the colon tissues. Differentially expressed genes in colon tissues were analyzed by transcriptomics. Cell apoptosis was detected by TUNEL staining. The expression of inflammatory factors such as IL-6 and IL-1β, as well as the expression of p-STAT1, p-JAK2, p-STAT3, Bax, and Bcl-2, were detected by immunofluorescence and immunohistochemistry. The expression of occludin, Bcl-2, Bax, and JAK/STAT signaling pathway-related vital proteins were quantified by Western blot (WB).
RESULTS: LRWE alleviated body weight loss, colon shortening, DAI scores, pathological changes, and ultrastructural features of colon tissue in mice with colitis. It also inhibited the increase of pro-inflammatory cytokines (such as TNF-α, IL-6, and IL-1β) and increased IL-10 levels. Additionally, it protected the intestinal barrier by upregulating the expression of Occludin and Mucin-2. Mechanistically, LRWE could inhibit the activation of JAK-STAT signaling pathway by reducing the protein expression of p-JAK2, p-STAT3, p-STAT1, Bcl2, and Bax, thus reducing the inflammatory responses and inhibiting cell apoptosis.
CONCLUSIONS: LRWE has a protective effect on DSS-induced UC. This effect is related to the inhibition of the JAK-STAT signaling pathway, the improvement of intestinal inflammation, and the reduction of intestinal epithelial cell apoptosis.