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张瑛,高晓峰,郭纯,张宇星,周德生.基于PPARγ信号通路探讨安脑平冲方极化M2型小胶质细胞减轻脑出血后神经炎症的作用机制[J].湖南中医药大学学报,2023,43(3):405-412[点击复制] |
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| 基于PPARγ信号通路探讨安脑平冲方极化M2型小胶质细胞减轻脑出血后神经炎症的作用机制 |
| 张瑛,高晓峰,郭纯,张宇星,周德生 |
| (湖南中医药大学研究生院, 湖南 长沙 410208;湖南中医药大学第一附属医院脑病一科, 湖南 长沙 410007;湖南中医药大学第一附属医院医学创新中心, 湖南 长沙 410007) |
| 摘要: |
| 目的 探讨安脑平冲方极化M2型小胶质细胞减轻脑出血（intracerebral hemorrhage, ICH）后神经炎症的作用机制。方法 采用氯化血红素干预BV2细胞模拟体外ICH模型。第一部分实验分3组[对照组、模型组、安脑平冲方含药血清组（A-HYXQ组）]，对照组为正常生长的BV2细胞，模型组及A-HYXQ组采用氯化血红素干预BV2细胞，24 h后模型组给予10%空白血清干预，A-HYXQ组给予10% A-HYXQ干预。采用免疫荧光和Western blot法检测Ym-1、白细胞介素-10（interleukin-10, IL-10）、诱导型一氧化氮合酶（iductible nitric oxide synthase, iNOS）、白细胞介素-6（interleukin-6, IL-6）表达量。第二部分实验分为对照组、模型组、A-HYXQ组、A-HYXQ+GW9662组，对照组、模型组、A-HYXQ组处理同第一部分实验，A-HYXQ+GW9662组在给予10% A-HYXQ前1 h给予10 μmol/L的GW9662，采用免疫荧光、Western blot和qRT-PCR法检测过氧化物酶体增殖物激活受体γ（peroxisome proliferators-activated receptors γ, PPARγ）、精氨酸酶-1（arginase-1, Arg-1）和白细胞介素-4（interleukin-4, IL-4）的表达量；采用Western blot、qRT-PCR和ELISA法检测白细胞介素-1β（interleukin-1β, IL-1β）和肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）。结果 第一部分：与对照组比较，模型组Ym-1和IL-10平均荧光强度和蛋白表达量明显下降（P＜0.05），而iNOS和IL-6平均荧光强度和蛋白表达量明显上升（P＜0.05）；与模型组相比，A-HYXQ组Ym-1和IL-10平均荧光强度和蛋白表达量明显上升（P＜0.05），而iNOS和IL-6平均荧光强度和蛋白表达量明显下降（P＜0.05）。第二部分：与对照组比较，模型组PPARγ、Arg-1和IL-4平均荧光强度、蛋白及mRNA表达量明显下降（P＜0.05），而TNF-α和IL-1β蛋白和上清液表达量明显上升（P＜0.05）；与模型组相比，A-HYXQ组PPARγ、Arg-1和IL-4平均荧光强度、蛋白及mRNA表达量明显上升（P＜0.05），而TNF-α和IL-1β蛋白和上清液表达量明显下降（P＜0.05）；与A-HYXQ组相比，A-HYXQ+GW9662组PPARγ、Arg-1和IL-4平均荧光强度、蛋白及mRNA表达量下降（P＜0.05），而TNF-α、IL-1β蛋白和上清液表达量上升（P＜0.05）。结论 安脑平冲方可能通过激活PPARγ信号通路极化M2型小胶质细胞，减轻氯化血红素干预BV2细胞的炎症反应。 |
| 关键词: 脑出血 神经炎症 小胶质细胞 过氧化物酶体增殖物激活受体γ 安脑平冲方 |
| DOI：10.3969/j.issn.1674-070X.2023.03.007 |
| 投稿时间：2022-07-07 |
| 基金项目:国家自然科学基金项目（81874463）；中医内科重大疾病防治研究与转化教育部重点实验室开放基金（ZYNK201609）；湖南省科技厅科技创新平台与人才计划（2017SK4005）；湖南省重点研发计划（2020SK2092）。 |
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| Mechanism of Annao Pingchong Formula on polarization of M2 microglia in alleviating neuroinflammation after intracerebral hemorrhage based on PPARγ signaling pathway |
| ZHANG Ying,GAO Xiaofeng,GUO Chun,ZHANG Yuxing,ZHOU Desheng |
| (Graduate School of Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Department of Encephalopath (I), The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Center for Medical Innovation, The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To explore the mechanism of Annao Pingchong Formula on polarization of M2 microglia in alleviating neuroinflammation after intracerebral hemorrhage (ICH). Methods The ICH model in vitro was simulated by the intervention of hemin on BV2 cells. The first part of the experiment (Part Ⅰ) was divided into three groups: control group, model group, and serum containing Annao Pingchong Formula group (A-HYXQ group). The control group was made up of BV2 cells with normal growth. The model group and A-HYXQ group were treated with hemin. Then after 24 h, the model group was treated with 10% blank serum, while the A-HYXQ group was treated with 10% A-HYXQ. Immunofluorescence and Western blot were used to detect the expression of Ym-1, interleukin-10 (IL-10), inducible nitric oxide synthase (iNOS) and interleukin-6 (IL-6). The second part (Part Ⅱ) of the experiment was divided into control group, model group, A-HYXQ group and A-HYXQ+GW9662 group. The treatment of the control, model and A-HYXQ groups was the same as that of Part Ⅰ. The A-HYXQ+GW9662 group was given GW9662 (10 μmol/L) 1 h before administration of 10% A-HYXQ. The expression of peroxisome proliferator-activated receptor-γ (PPAR γ), arginase-1 (Arg-1) and interleukin-4 (IL-4) was detected by immunofluorescence, Western blot and qRT-PCR, while tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by Western blot, qRT-PCR and ELISA. Results Part Ⅰ: Compared with the control group, the average fluorescence intensity and protein expression of Ym-1 and IL-10 in the model group were reduced significantly (P<0.05), while those of iNOS and IL-6 were significantly higher (P<0.05); compared with the model group, the average fluorescence intensity and protein expression of Ym-1 and IL-10 in the A-HYXQ group were significantly higher (P<0.05), while those of iNOS and IL-6 were reduced significantly (P<0.05). Part Ⅱ: Compared with the control group, the average fluorescence intensity, protein and mRNA expression of PPARγ, Arg-1 and IL-4 in the model group were significantly lower (P<0.05), however, the protein expression of TNF-α and IL-1β and the supernatant expression were significantly higher (P<0.05); compared with the model group, the average fluorescence intensity, protein and mRNA expression of PPARγ, Arg-1 and IL-4 in the A-HYXQ group were significantly higher (P<0.05), while the protein expression of TNF-α and IL-1β and the supernatant expression were reduced significantly (P<0.05); compared with the A-HYXQ group, the average fluorescence intensity, protein and mRNA expression of PPARγ, Arg-1 and IL-4 in the A-HYXQ+GW9662 group were reduced significantly (P<0.05), while the protein expression of TNF-α, IL-1β and the supernatant expression were significantly higher (P<0.05). Conclusion Annao Pingchong Formula may activate PPARγ signaling pathway to polarize M2-microglia, thus reducing the inflammatory reaction of BV2 cells after hemin intervention. |
| Key words: intracerebral hemorrhage neuroinflammation microglia peroxisome proliferators-activated receptors-γ Annao Pingchong Formula |
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