含益气复脉通络方的血清通过MAPK通路减轻高糖诱导的小胶质细胞损伤基于网络药理学的分析和生物学验证

IF 1.9 4区医学 Q3 INTEGRATIVE & COMPLEMENTARY MEDICINE

European Journal of Integrative Medicine Pub Date : 2024-09-08 DOI:10.1016/j.eujim.2024.102403

Chang Chang , Qian Chen , Ke Zuo , Huilun Yuan , Lujie Wang , Aijuan Jiang , Yuqing Wang

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引用次数: 0

摘要

导言益气活血通络方（YQHX）由黄芪（Astragalus mongholicus Bunge）、当归（Angelica sinensis (Oliv.) Diels）、熟地黄（Rehmannia glutinosa Libosch、YQHX是安徽中医药大学第一附属医院的中成药方剂，可减轻糖尿病神经病变疼痛（DNP）。本研究旨在探讨 YQHX 介导的 DNP 缓解机制。方法采用液相色谱串联质谱法（LC-MS/MS）鉴定YQHX中的化学成分。通过网络药理学分析探讨了 YQHX 治疗 DNP 的潜在机制。用不同浓度的含 YQHX 的血清处理已建立的葡萄糖诱导 BV2 细胞损伤模型。细胞计数试剂盒-8和酶联免疫吸附试验评估了含YQHX的血清和MAPK通路抑制剂对小胶质细胞的影响。测定了小胶质细胞标志物 OX42 的信使 RNA (mRNA) 和蛋白水平。此外，还通过 Western 印迹法检测了磷酸化-p38、-c-Jun-terminal 激酶和-细胞外信号调节激酶以及下游核因子-κB/p65 和磷酸化-激活转录因子 2 (ATF2) 蛋白。网络药理学结果显示，203 种有效成分和 363 个重叠靶点分别是 YQHX 介导的 DNP 治疗的潜在候选疗法。此外，京都基因和基因组百科全书的通路富集分析共得出 105 个信号通路。值得注意的是，YQHX大大提高了BV2细胞的活力（P <0.01），显著减少了促炎因子的产生（P <0.01），显著增加了抗炎因子的产生（P <0.01），并明显抑制了OX42的mRNA和蛋白水平（P <0.01）。结论YQHX可通过抑制MAPKs和下游ATF2的活化来防止高血糖诱导的BV2微神经胶质细胞损伤，从而减少微神经胶质细胞的活化和促炎细胞因子的释放。这些结果为进一步探索YQHX治疗DNP的机制提供了基础理论依据，也为中药的应用提供了一个新的靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Yiqi Huoxue Tongluo formula-containing serum attenuates high glucose-induced injury in microglia via MAPK pathways: Network pharmacology-based analysis and biological validation

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Yiqi Huoxue Tongluo formula-containing serum attenuates high glucose-induced injury in microglia via MAPK pathways: Network pharmacology-based analysis and biological validation

Introduction

The Yiqi Huoxue Tongluo formula (YQHX), containing Astragalus mongholicus Bunge, Angelica sinensis (Oliv.) Diels, Rehmannia glutinosa Libosch., Pueraria lobata (Willd.) Ohwi, Corydalis yanhusuo W.T.Wang, Spatholobus suberectus Dunn, Clematis chinensis Osbeck, is a proprietary formula of the First Affiliated Hospital of Anhui University of Chinese Medicine, and can attenuate diabetic neuropathy pain (DNP). This study aimed to explore the mechanisms underlying YQHX-mediated DNP alleviation. Additionally, the effects of YQHX on BV2 microglia injured by high glucose via the mitogen-activated protein kinase (MAPK) signaling pathways were investigated.

Methods

Liquid chromatography tandem mass spectroscopy (LC-MS/MS) was performed to identify the chemical compounds in YQHX. The potential mechanism of YQHX for treating DNP was explored through network pharmacology analysis. The established glucose-induced BV2 cell injury model was treated with varying concentrations of YQHX-containing serum. The effects of YQHX-containing serum and MAPK pathway inhibitors on microglial cells were assessed by Cell Counting Kit-8 and enzyme-linked immunosorbent assays. The messenger RNA (mRNA) and protein levels of microglial marker OX42 were measured. Additionally, phosphorylated-p38, -c-Jun-terminal kinase, and -extracellular signal-regulated kinase and downstream nuclear factor-κB/p65 and phosphorylated-activating transcription factor 2 (ATF2) proteins were detected by western blotting.

Results

In total, 503 chemical components were identified in YQHX by LC-MS/MS analysis. Network pharmacology results showed 203 active ingredients and 363 overlapping targets, respectively, as potential therapeutic candidates for YQHX-mediated DNP treatment. Additionally, the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis yielded a total of 105 signal pathways. Notably, YQHX considerably improved the cell viability of BV2 cells (P < 0.01), remarkably reduced the production of proinflammatory factors (P < 0.01), significantly increased the production of anti-inflammatory factors (P < 0.01), and markedly inhibited the mRNA and protein levels of OX42 (P < 0.01). Furthermore, YQHX treatment inhibited MAPK activation and ATF2 expression.

Conclusions

YQHX may protect against high-glucose-induced BV2 microglial cell injury by inhibiting the activation of MAPKs and downstream ATF2, thus, reducing microglial activation and proinflammatory cytokine release. These results provide a foundational theoretical basis for exploring the further mechanism of YQHX in treating DNP, and they provide a novel target for the application of traditional Chinese medicine.

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来源期刊

European Journal of Integrative Medicine INTEGRATIVE & COMPLEMENTARY MEDICINE-

CiteScore

4.70

自引率

4.00%

发文量

102

审稿时长

33 days

期刊介绍： The European Journal of Integrative Medicine (EuJIM) considers manuscripts from a wide range of complementary and integrative health care disciplines, with a particular focus on whole systems approaches, public health, self management and traditional medical systems. The journal strives to connect conventional medicine and evidence based complementary medicine. We encourage submissions reporting research with relevance for integrative clinical practice and interprofessional education. EuJIM aims to be of interest to both conventional and integrative audiences, including healthcare practitioners, researchers, health care organisations, educationalists, and all those who seek objective and critical information on integrative medicine. To achieve this aim EuJIM provides an innovative international and interdisciplinary platform linking researchers and clinicians. The journal focuses primarily on original research articles including systematic reviews, randomized controlled trials, other clinical studies, qualitative, observational and epidemiological studies. In addition we welcome short reviews, opinion articles and contributions relating to health services and policy, health economics and psychology.