School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China.
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China.
J Ethnopharmacol. 2022 Oct 5;296:115447. doi: 10.1016/j.jep.2022.115447. Epub 2022 Jun 8.
Hypericum sampsonii Hance (Yuanbaocao), a traditional herbal medicine with various pharmacological properties, is traditionally used to treat diarrhea and enteritis in China for hundreds of years. Investigations have uncovered its anti-inflammatory effects and corresponding bioactive constituents in H. sampsonii, however, the mechanisms of action for the treatment of enteritis are still unclear.
This study aims to investigate the therapeutic effects and molecular mechanisms of H. sampsonii in a dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) mice model.
The major ingredients of the ethyl acetate extract (HS) in H. sampsonii were analyzed by UPLC-QTOF-MS. The inflammatory state of UC mice was caused by 3% DSS once daily for seven days. During DSS treatment, the mice in the positive drug group and the other three groups were orally administered 5-ASA (positive control) or HS daily. After treatment with HS or 5-ASA for a week, colonic pathological observation and the molecular biological index were performed for therapeutic evaluation, including visual inspection in the length and weight of colons and spleens, pathological morphology by hematoxylin and eosin (HE) staining, determination of oxidative markers, inflammatory cytokines and tumor necrosis factor-alpha (TNF-α) levels in colonic tissues as well as spleen index. Gene expression levels of inflammatory cytokines, antioxidant enzymes and PDE4 were detected using kits and PCR, while the expression of colonic tight junction proteins and relative signals of PKA/CREB signaling pathway were analyzed by Western blot.
The main components in HS were found to be polycyclic polyprenylated acylphloroglucinols (PPAPs). HS distinctly alleviated DSS-stimulated UC-like lesions symptoms as evidenced by a significant recovery from body weight, colon lengths, and histological injuries of colons. HS reduced the accumulation of pro-inflammatory cytokines and improved the mRNA level of IL-10. Simultaneously, the colonic mRNA expression levels of IL-1β, IL-17, iNOS and COX-2 were all significantly suppressed by HS in a dose-dependent manner. Furthermore, HS restored the protein expression of tight junction-associated protein (ZO-1 and occluding). Besides, HS significantly inhibited the protein level of PDE4 and decreased the expressions of PKA and phosphorylated CREB.
This is the first work about main composition and anti-UC effect of Hypericum sampsonii Hance. For the first time, this study reveals HS is not toxic in a single dose and exert significantly protective effect in DSS-colitis mice. The underlying mechanisms may involve the improvement to inflammatory status, the protection for intestinal barrier function, the inhibition of PDE4, and the activation of PKA/CREB signaling pathway. This study provided an experimental basis for the traditional application of H. sampsonii Hance in the treatment of diarrhea and dysentery.
贯叶金丝桃（元宝草）是一种具有多种药理特性的传统草药，在中国已有数百年用于治疗腹泻和肠炎的历史。研究已经揭示了贯叶金丝桃中的抗炎作用及其相应的生物活性成分，然而，其治疗肠炎的作用机制仍不清楚。
本研究旨在探讨贯叶金丝桃在葡聚糖硫酸钠（DSS）诱导的溃疡性结肠炎（UC）小鼠模型中的治疗效果和分子机制。
采用 UPLC-QTOF-MS 分析贯叶金丝桃中乙酸乙酯提取物（HS）的主要成分。通过每天给予 3% DSS 一次，持续 7 天，诱导 UC 小鼠的炎症状态。在 DSS 处理期间，阳性药物组和其他三组小鼠每天口服 5-ASA（阳性对照）或 HS 进行治疗。用 HS 或 5-ASA 治疗一周后，进行结肠组织的病理观察和分子生物学指标评价，包括结肠和脾脏长度和重量的肉眼观察、苏木精和伊红（HE）染色的病理形态学、氧化标志物、炎性细胞因子和肿瘤坏死因子-α（TNF-α）水平的测定以及结肠组织中炎性细胞因子、抗氧化酶和 PDE4 的基因表达水平，通过 Western blot 分析结肠紧密连接蛋白的表达和 PKA/CREB 信号通路的相对信号。
发现 HS 中的主要成分是多环多聚缩酚酸内酯（PPAPs）。HS 明显缓解了 DSS 刺激的 UC 样病变症状，表现在体重、结肠长度和结肠组织学损伤的显著恢复。HS 减少了促炎细胞因子的积累，并提高了 IL-10 的 mRNA 水平。同时，HS 以剂量依赖性方式显著抑制了 IL-1β、IL-17、iNOS 和 COX-2 的结肠组织 mRNA 表达水平。此外，HS 还显著恢复了紧密连接相关蛋白（ZO-1 和闭合蛋白）的蛋白表达。此外，HS 显著抑制了 PDE4 的蛋白水平，并降低了 PKA 和磷酸化 CREB 的表达。
这是首次关于贯叶金丝桃主要成分和抗 UC 作用的研究。本研究首次揭示，HS 在单剂量下无毒性，并在 DSS-结肠炎小鼠中表现出显著的保护作用。其潜在机制可能涉及改善炎症状态、保护肠道屏障功能、抑制 PDE4 和激活 PKA/CREB 信号通路。本研究为贯叶金丝桃在治疗腹泻和痢疾方面的传统应用提供了实验依据。
