Penehyclidine hydrochloride alleviates LPS-induced inflammatory responses and oxidative stress via ROS/Nrf2/HO-1 activation in RAW264.7 cells.

IF 2.1 4区医学 Q3 MEDICINE, RESEARCH & EXPERIMENTAL

Advances in Clinical and Experimental Medicine Pub Date : 2024-03-26 DOI:10.17219/acem/183883

Qiongmei Guo, Chunyan Zhang, Jingui Gao, Wenjing Shi, Xiaozhi Liu

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引用次数: 0

Abstract

Background: Inflammation is a biological response of the immune system to harmful stimuli. Penehyclidine hydrochloride (PCH) can alleviate inflammation and oxidative stress by activating reactive oxygen species (ROS), nuclear factor erythroid 2-related factor (Nrf2) and heme oxygenase 1 (HO-1) in animal models, but there is a lack of cellular evidence.

Objectives: This study investigated the effects of PHC on lipopolysaccharide (LPS)-induced inflammation response and oxidative stress in RAW264.7 cells.

Material and methods: RAW264.7 cells were treated with 1 μg/mL or 5 μg/mL of PHC, with interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), IL-1β, and prostaglandin E2 (PGE2) levels measured with enzyme-linked immunosorbent assay (ELISA) and nitric oxide (NO) measured using the Griess test. Reactive oxygen species were examined with flow cytometry and immunofluorescence, and b-related factor 2 (BRF-2) and NAD(P)H-quinone oxidoreductase 1 (NQO1) using western blot.

Results: Penehyclidine hydrochloride partly, but substantially, reversed LPS-related NO and PGE2 production by RAW264.7 cells in a dose-dependent manner and suppressed LPS-induced expression of IL-6, TNF-α and IL-1β messenger ribonucleic acid (mRNA), secretion of IL-6, TNF-α and IL-1β, and ROS production. Lipopolysaccharide stimulation did not affect Nrf2, heme oxygenase 1 (HO-1) or NQO1 protein expression in RWA264.7 cells not treated with PHC. However, PHC treatment significantly elevated Nrf2, HO-1 and NQO1 protein in LPS-treated RWA264.7 cells, an effect that was dose-dependent. The ROS scavenging using N-acetyl-L-cysteine abolished the PHC-induced upregulation of Nrf2 and HO-1.

Conclusions: Penehyclidine hydrochloride may alleviate LPS-induced inflammation and oxidative stress by activating Nrf2 signaling in RAW264.7 macrophages. These findings suggest that PHC could alleviate inflammation by targeting activated macrophages.

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盐酸哌替啶通过激活 ROS/Nrf2/HO-1 减轻 RAW264.7 细胞中 LPS 诱导的炎症反应和氧化应激。

背景：炎症是免疫系统对有害刺激的一种生物反应。在动物模型中，盐酸哌替啶（PCH）可通过激活活性氧（ROS）、核因子红细胞2相关因子（Nrf2）和血红素加氧酶1（HO-1）来缓解炎症和氧化应激，但缺乏细胞证据：本研究探讨了PHC对脂多糖（LPS）诱导的RAW264.7细胞炎症反应和氧化应激的影响：用1 μg/mL或5 μg/mL的PHC处理RAW264.7细胞，用酶联免疫吸附试验（ELISA）检测白细胞介素6（IL-6）、肿瘤坏死因子α（TNF-α）、IL-1β和前列腺素E2（PGE2）的水平，用Griess试验检测一氧化氮（NO）的水平。用流式细胞仪和免疫荧光法检测了活性氧，用 Western 印迹法检测了 b 相关因子 2（BRF-2）和 NAD(P)H-醌氧化还原酶 1（NQO1）：结果：盐酸哌替啶以剂量依赖的方式部分但显著地逆转了 RAW264.7 细胞与 LPS 相关的 NO 和 PGE2 的产生，并抑制了 LPS 诱导的 IL-6、TNF-α 和 IL-1β 信使核糖核酸（mRNA）的表达、IL-6、TNF-α 和 IL-1β 的分泌以及 ROS 的产生。在未经 PHC 处理的 RWA264.7 细胞中，脂多糖刺激不会影响 Nrf2、血红素加氧酶 1（HO-1）或 NQO1 蛋白的表达。然而，在经 LPS 处理的 RWA264.7 细胞中，PHC 能明显提高 Nrf2、HO-1 和 NQO1 蛋白的表达，这种影响与剂量有关。使用 N-乙酰-L-半胱氨酸清除 ROS 可消除 PHC 诱导的 Nrf2 和 HO-1 上调：结论：盐酸哌替啶可通过激活 RAW264.7 巨噬细胞中的 Nrf2 信号，减轻 LPS 诱导的炎症和氧化应激。这些研究结果表明，PHC 可通过靶向活化的巨噬细胞来缓解炎症。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Advances in Clinical and Experimental Medicine MEDICINE, RESEARCH & EXPERIMENTAL-

CiteScore

3.70

自引率

4.80%

发文量

153

审稿时长

6-12 weeks

期刊介绍： Advances in Clinical and Experimental Medicine has been published by the Wroclaw Medical University since 1992. Establishing the medical journal was the idea of Prof. Bogumił Halawa, Chair of the Department of Cardiology, and was fully supported by the Rector of Wroclaw Medical University, Prof. Zbigniew Knapik. Prof. Halawa was also the first editor-in-chief, between 1992-1997. The journal, then entitled "Postępy Medycyny Klinicznej i Doświadczalnej", appeared quarterly. Prof. Leszek Paradowski was editor-in-chief from 1997-1999. In 1998 he initiated alterations in the profile and cover design of the journal which were accepted by the Editorial Board. The title was changed to Advances in Clinical and Experimental Medicine. Articles in English were welcomed. A number of outstanding representatives of medical science from Poland and abroad were invited to participate in the newly established International Editorial Staff. Prof. Antonina Harłozińska-Szmyrka was editor-in-chief in years 2000-2005, in years 2006-2007 once again prof. Leszek Paradowski and prof. Maria Podolak-Dawidziak was editor-in-chief in years 2008-2016. Since 2017 the editor-in chief is prof. Maciej Bagłaj. Since July 2005, original papers have been published only in English. Case reports are no longer accepted. The manuscripts are reviewed by two independent reviewers and a statistical reviewer, and English texts are proofread by a native speaker. The journal has been indexed in several databases: Scopus, Ulrich’sTM International Periodicals Directory, Index Copernicus and since 2007 in Thomson Reuters databases: Science Citation Index Expanded i Journal Citation Reports/Science Edition. In 2010 the journal obtained Impact Factor which is now 1.179 pts. Articles published in the journal are worth 15 points among Polish journals according to the Polish Committee for Scientific Research and 169.43 points according to the Index Copernicus. Since November 7, 2012, Advances in Clinical and Experimental Medicine has been indexed and included in National Library of Medicine’s MEDLINE database. English abstracts printed in the journal are included and searchable using PubMed http://www.ncbi.nlm.nih.gov/pubmed.