香连丸通过 TLR4/MyD88/NF-κB 信号通路减轻溃疡性结肠炎。
Xianglian Pill attenuates ulcerative colitis through TLR4/MyD88/NF-κB signaling pathway.
机构信息
School of Chinese Medicine, School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China; The First School of Clinical Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
Department of Colorectal Surgery, Kunshan Hospital of Traditional Chinese Medicine, Suzhou, 215300, China.
出版信息
J Ethnopharmacol. 2023 Jan 10;300:115690. doi: 10.1016/j.jep.2022.115690. Epub 2022 Sep 6.
ETHNOPHARMACOLOGICAL RELEVANCE
Xianglian Pill (XLP) is a classical Chinese medicine prescription applied for controlling ulcerative colitis (UC). Whereas, the underlying mechanism remains unclear.
AIM OF THE STUDY
The present work was aimed to investigate the mechanism of XLP in dextran sulfate sodium (DSS)-induced UC via the Toll Like Receptor 4 (TLR4)/Myeloid Differentiation factor 88 (MyD88)/Nuclear Factor kappa-B (NF-κB) signaling in mice.
MATERIALS AND METHODS
The major components of XLP were detected by high-performance liquid chromatography-diode array detection (HPLC-DAD). The ulcerative colitis model was induced by DSS in mice. 5-Amino Salicylic Acid (5-ASA) group and XLP group were intragastrically treated. Disease activity index (DAI) and colon length were monitored and hematoxylin-eosin (HE) staining was conducted. Gasdermin D (GSDMD)-N and TLR4 expressions in colon tissues were visualized by immunofluorescence. TLR4 mRNA was measured by Real Time Quantitative PCR (RT-qPCR). The expressions of NOD-like receptor thermal protein domain associated protein 3 (NLRP3), active-caspase-1, GSDMD-N, TLR4, MYD88, NF-κB, p-NF-κB, and the ubiquitination of TLR4 in colon tissues were detected by Western blot. Myeloperoxidase (MPO) enzyme activity was examined and serum inflammatory factors Interleukin (IL)-1β, IL-6, Tumor Necrosis Factor-α (TNF-α), and IL-18 were determined by Enzyme-linked Immunosorbent Assay (ELISA). TLR4 mice were applied for verifying the mechanism of XLP attenuated DSS symptoms.
RESULTS
The XLP treatment extended colon length, reduced DAI, and attenuated histopathological alteration in DSS-induced mice. XLP administration suppressed MPO activity and reduced the content of IL-1β, IL-6, TNF-α and IL-18 in serum. XLP also inhibited the expression levels of GSDMD-N, TLR4, NLRP3, active-caspase-1, MyD88, p-NF-κB/NF-κB in colon tissues of DSS-induced mice. TLR4 mice proved that TLR4 was involved in XLP-mediated beneficial effect on DSS-induced ulcerative colitis.
CONCLUSIONS
XLP might treat ulcerative colitis by regulating the TLR4/MyD88/NF-κB signaling pathway.
民族药理学相关性
香连丸（XLP）是一种用于控制溃疡性结肠炎（UC）的经典中药方剂。然而，其潜在机制尚不清楚。
研究目的
本研究旨在通过 Toll 样受体 4（TLR4）/髓样分化因子 88（MyD88）/核因子 kappa-B（NF-κB）信号通路，探讨 XLP 对葡聚糖硫酸钠（DSS）诱导的 UC 小鼠的作用机制。
材料和方法
采用高效液相色谱-二极管阵列检测法（HPLC-DAD）检测 XLP 的主要成分。采用 DSS 诱导小鼠溃疡性结肠炎模型。5-氨基水杨酸（5-ASA）组和 XLP 组进行灌胃治疗。监测疾病活动指数（DAI）和结肠长度，进行苏木精-伊红（HE）染色。免疫荧光法观察结肠组织中 Gasdermin D（GSDMD）-N 和 TLR4 的表达。实时定量 PCR（RT-qPCR）法检测 TLR4mRNA 的表达。Western blot 法检测结肠组织中 NOD 样受体热蛋白结构域相关蛋白 3（NLRP3）、活化半胱天冬酶-1、GSDMD-N、TLR4、MyD88、NF-κB、p-NF-κB 和 TLR4 泛素化的表达。采用髓过氧化物酶（MPO）酶活性检测和酶联免疫吸附试验（ELISA）法检测血清中白细胞介素（IL）-1β、IL-6、肿瘤坏死因子-α（TNF-α）和 IL-18 等炎症因子的含量。采用 TLR4 小鼠验证 XLP 减轻 DSS 症状的作用机制。
结果
XLP 治疗可延长结肠长度，降低 DAI，并减轻 DSS 诱导的小鼠组织病理学改变。XLP 给药可抑制 MPO 活性，降低血清中 IL-1β、IL-6、TNF-α和 IL-18 的含量。XLP 还可抑制 DSS 诱导的小鼠结肠组织中 GSDMD-N、TLR4、NLRP3、活化半胱天冬酶-1、MyD88、p-NF-κB/NF-κB 的表达水平。TLR4 小鼠证实 TLR4 参与了 XLP 对 DSS 诱导的溃疡性结肠炎的有益作用。
结论
XLP 可能通过调节 TLR4/MyD88/NF-κB 信号通路治疗溃疡性结肠炎。
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