白头翁汤通过抑制TLR4-ERK1/2信号通路缓解LPS诱导的微血管内皮细胞炎性反应

doi:10.11843/j.issn.0366-6964.2021.08.025

摘要/Abstract

摘要： 近年来，中药复方抗炎机制的研究不断深入，白头翁汤复方（Pulsatilla decoction，PD）作为经典的清热解毒中药方剂常用于预防和治疗细菌性腹泻。然而其抗炎机制和靶细胞研究仍然不明确，本课题以大鼠肠黏膜微血管内皮细胞（RIMVECs）为模式细胞，旨在研究白头翁汤对LPS诱导的RIMVECs炎症反应的调控作用。利用LPS刺激RIMVECs，通过荧光定量PCR（RT-PCR）、蛋白免疫印迹（Western blot）的方法检测白头翁汤对LPS刺激后RIMVECs的炎性信号通路TLR4-ERK1/2信号通路关键蛋白TLR4、TRAF6、ERK的mRNA及蛋白表达。进一步采用酶联免疫吸附方法（ELISA）检测白头翁汤对LPS刺激后的炎性因子IL-6、IL-8、IL-1β、TNF-α的分泌情况。结果表明：白头翁汤可以显著降低LPS诱导的TLR4、TRAF6、ERK的mRNA水平及蛋白表达并降低了LPS诱导的细胞下游炎性因子的分泌。白头翁汤通过抑制TLR4-ERK1/2信号通路缓解LPS所诱导的RIMVECs炎性反应，发挥抗炎作用。

关键词: 微血管内皮细胞, LPS, 白头翁汤, ERK1/2信号通路, PD98059

Abstract: In recent years, the researches on the anti-inflammatory mechanisms of Traditional Chinese Medicine formula have been in-depth. Pulsatilla decoction served as a classic Traditional Chinese Medicine prescription for clearing away heat and toxic material, is usually employed to prevent and treat bacterial diarrhea. However, its anti-inflammatory mechanisms and target cells are still unclear. In this study, rat intestinal mucosa microvascular endothelial cells (RIMVECs) were used as the model cells, aiming to illustrate the regulation effects of Pulsatilla decoction on LPS-induced inflammation of RIMVECs. The levels of mRNA and protein expressions of key proteins TLR4, TRAF6 and ERK in the inflammatory signaling pathway of RIMVECs after LPS stimulation were detected by RT-PCR and Western blot. The release levels of IL-6, IL-8, IL-1β, and TNF-alpha were determined by ELISA. The results showed that Pulsatilla decoction could significantly reduce the mRNA levels and protein expressions of TLR4, TRAF6 and ERK induced by LPS, and also suppress the secretions of the downstream inflammatory factors induced by LPS. Pulsatilla decoction alleviates the inflammatory responses of LPS-induced RIMVECs by inhibiting TLR4-ERK1/2 signaling pathway and plays an anti-inflammatory role.

Key words: microvascular endothelial cells, LPS, Pulsatilla decoction, ERK1/2 signaling pathway, PD98059

中图分类号:

S853.91

贺尚文, 王月明, 张慧, 侯思鲁, 刘晓晔, 董虹. 白头翁汤通过抑制TLR4-ERK1/2信号通路缓解LPS诱导的微血管内皮细胞炎性反应[J]. 畜牧兽医学报, 2021, 52(8): 2317-2325.

HE Shangwen, WANG Yueming, ZHANG Hui, HOU Silu, LIU Xiaoye, DONG Hong. Pulsatilla Decoction Alleviates LPS-induced Inflammation of Microvascular Endothelial Cells by Inhibiting the TLR4-ERK1/2 Signaling Pathway[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(8): 2317-2325.

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