毛樱桃总黄酮对RAW264.7细胞中炎症因子水平的影响及其机制

doi:10.13481/j.1671-587x.20190416

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (04): 836-842.doi: 10.13481/j.1671-587x.20190416

• 基础研究 • 上一篇

毛樱桃总黄酮对RAW264.7细胞中炎症因子水平的影响及其机制

曹爽¹, 范紫薇¹, 王映映², 孙利娟², 古虹¹, 李贺¹, 孙靖辉¹, 王春梅¹, 陈建光¹, 陈曦², 张成义¹

1. 北华大学药学院药理教研室, 吉林吉林 132013;
2. 北华大学医学院病原学教研室, 吉林吉林 132013

收稿日期:2018-09-05 发布日期:2019-08-02
通讯作者: 张成义,教授,硕士研究生导师(Tel:0432-64608281,E-mail:zhchyJL@163.com);陈曦,教授,硕士研究生导师(Tel:0432-64608281,E-mail:846617750@qq.com) E-mail:zhchyJL@163.com;846617750@qq.com
作者简介:曹爽(1994-),女,内蒙古自治区乌兰浩特市人,在读医学硕士,主要从事抗炎药物方面的研究。
基金资助:
吉林省科技厅科技攻关项目资助课题（20190304048YY）；吉林省吉林市科技局科技计划项目资助课题（201820847）；吉林省中医药管理局中医药科技项目资助课题（2018111）

Effects of prunus tomentosa thunb total flavonoids on levels of inflammatory factors in RAW264.7 cells and their mechanisms

CAO Shuang¹, FAN Ziwei¹, WANG Yingying², SUN Lijuan², GU Hong¹, LI He¹, SUN Jinghui¹, WANG Chunmei¹, CHEN Jianguang¹, CHEN Xi², ZHANG Chengyi¹

1. Department of Pharmacology, College of Pharmacy, Beihua University, Jilin 132013, China;
2. Department of Etiology, College of Medical, Beihua University, Jilin 132013, China

Received:2018-09-05 Published:2019-08-02

摘要/Abstract

摘要： 目的：探讨毛樱桃总黄酮（PTTTF）对脂多糖（LPS）诱导的细胞炎症模型中炎症因子的影响，阐明其作用机制。方法：通过LPS诱导小鼠巨噬细胞RAW264.7和人中性粒细胞（PMN）建立细胞炎症模型，将细胞分为对照组、模型组、地塞米松（DXM）组和不同剂量（0.4、4.0和40.0 mg·L^-1）PTTTF组。采用RT-PCR法检测RAW264.7细胞中白细胞介素1β（IL-1β）、白细胞介素6（IL-6）、诱导性一氧化氮合酶（iNOS）、肿瘤坏死因子（TNF-α）和高迁移率族蛋白B1（HMGB1）mRNA表达水平；采用Western blotting法检测RAW264.7细胞中核转录因子κB（NF-κB）及信号通路关键分子细胞外调节蛋白激酶（ERK1/2）、C-Jun氨基末端激酶1/2（JNK）和磷酸化JNK（p-JNK）蛋白表达水平；采用流式细胞术检测RAW264.7细胞S期细胞百分率和PMN细胞凋亡率。结果：与对照组比较，模型组RAW264.7细胞中IL-1β、IL-6、iNOS、TNF-α和HMGB1 mRNA表达水平明显升高（P<0.05或P<0.01），NF-κB、ERK1/2和p-JNK蛋白表达水平明显升高（P<0.01），S期细胞百分率明显升高（P<0.05），PMN细胞凋亡百分率明显降低（P<0.05）。与模型组比较，4.0及40.0 mg·L^-1PTTTF组和DXM组RAW264.7细胞中IL-1β、iNOS、TNF-α和HMGB1 mRNA表达水平明显降低（P<0.05或P<0.01），NF-κB、ERK1/2和p-JNK蛋白表达水平明显降低（P<0.05），S期细胞百分率明显降低（P<0.05或P<0.01），PMN细胞凋亡率明显升高（P<0.05或P<0.01）。结论：PTTTF能够下调LPS诱导的细胞炎症模型中炎症因子表达水平，该作用可能与其降低S期细胞百分率，促进其凋亡，并下调NF-κB、ERK1/2和p-JNK蛋白表达有关。

关键词: 毛樱桃总黄酮, 炎症因子, 信号通路, 细胞凋亡

Abstract: Objective:To investigate the influence of Prunus tomentosa thunb total flavonoids (PTTTF) in the inflammatory factors in the lipopolysaccharide(LPS)-induced cellular inflammation model, and to clarify its mechanism. Methods:The cellular inflammation models of the RAW264.7 cells and human neutrophils (PMN) induced by LPS were established. The cells were divided into control group, model group, dexamethasone group (DXM), and different doses (0.4,4.0, and 40.0 mg·L^-1) of PTTTF groups. The expression levels of inflammatory factors including interleukin-1β(IL-1β), interleukin-6(IL-6), inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α) and high mobility group protein B1(HMGB1)mRNA in the RAW264.7 cells were detected by RT-PCR. The expression levels of NF-κB, and signal pathway key molecules ERK1/2 and p-JNK proteins in the RAW264.7 cells were determined by Western blotting method. Flow cytometry was used to detect the percentages of RAW264.7 cells in different cell cycles and the apoptotic rates of PMN. Results:Compared with control group, the expression levels of IL-1β, IL-6, iNOS, TNF-α, and HMGB1 mRNA in the RAW264.7 cells in model group were significantly increased (P<0.05 or P<0.01),and the expression levels of NF-κB,ERK1/2, and p-JNK proteins were also significantly increased (P<0.01);the percentage of RAW264.7 cells in S phase was increased (P<0.05);the apoptotic rate of PMN was significantly decreased (P<0.05). Compared with model group,the expression levels of IL-1β, iNOS, TNF-α, and HMGB1 mRNA in the RAW264.7 cells in 4.0 and 40.0 mg·L^-1 PTTTF groups and DXM group were significantly decreased(P<0.05 or P<0.01), the expression levels of NF-κB,ERK1/2, and p-JNK proteins were significantly decreased(P<0.05), and the percentages of RAW264.7 cells in S phase were significantly decreased (P<0.05 or P<0.01); the apoptotic rates of PMN were increased(P<0.05 or P<0.01). Conclusion:PTTTF can down-regulate the expression levels of inflammatory factors in the RAW264.7 cell inflammation models induced by LPS. This effect may be related to decreasing the percentage of inflammatory cells in S phase, promoting the apoptosis, and down-regulating the expressions of NF-κB, ERK1/2, and p-JNK proteins.

Key words: prunus tomentosa thunb total flavonoids, inflammatory factors, signaling pathway, apoptosis

中图分类号:

R285.5

曹爽, 范紫薇, 王映映, 孙利娟, 古虹, 李贺, 孙靖辉, 王春梅, 陈建光, 陈曦, 张成义. 毛樱桃总黄酮对RAW264.7细胞中炎症因子水平的影响及其机制[J]. 吉林大学学报(医学版), 2019, 45(04): 836-842.

CAO Shuang, FAN Ziwei, WANG Yingying, SUN Lijuan, GU Hong, LI He, SUN Jinghui, WANG Chunmei, CHEN Jianguang, CHEN Xi, ZHANG Chengyi. Effects of prunus tomentosa thunb total flavonoids on levels of inflammatory factors in RAW264.7 cells and their mechanisms[J]. Journal of Jilin University(Medicine Edition), 2019, 45(04): 836-842.

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毛樱桃总黄酮对RAW264.7细胞中炎症因子水平的影响及其机制

Effects of prunus tomentosa thunb total flavonoids on levels of inflammatory factors in RAW264.7 cells and their mechanisms

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