| 甘海燕,李琳,杨琰,等.补阳还五汤调控小胶质细胞/巨噬细胞极化抑制大鼠脑缺血后炎症反应研究[J].浙江中医药大学学报,2019,43(1):1-6. |
| 补阳还五汤调控小胶质细胞/巨噬细胞极化抑制大鼠脑缺血后炎症反应研究 |
| Buyang Huanwu Decoction Inhibits Inflammation via Regulating Microglia/Macrophage Polarization after Cerebral Ischemia in Rats |
| DOI：10.16466/j.issn1005-5509.2019.01.001 |
| 中文关键词: 补阳还五汤 小胶质细胞/巨噬细胞 极化 炎症 脑缺血 |
| 英文关键词: Buyang Huanwu Decoction microglia/macrophage polarization inflammation cerebral ischemia |
| 基金项目:国家自然科学基金项目(81073075);浙江中医药大学校级科研基金项目(20172Y01 ) |
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| 中文摘要: |
| [目的]研究补阳还五汤(Buyang Huanwu Decoction, BYHWD)对大鼠脑缺血后小胶质细胞/巨噬细胞M1/M2极化及神经炎症的影响。[方法]采用线栓法建立大鼠大脑中动脉阻塞(middle cerebral artery occlusion, MCAO)模型，缺血90 min后再灌注。将大鼠随机分为假手术组、模型组和BYHWD组，BYHWD组缺血后24h开始予BYHWD(13g·kg-1)灌胃，连续给药14d。缺血后第14天处死大鼠，分别采用Iba1/CD16/32、Iba1/CD206免疫荧光双标染色检测缺血区M1型和M2型小胶质细胞/巨噬细胞表型，qRT-PCR检测M1型小胶质细胞/巨噬细胞表面标记物CD86、诱导型一氧化氮合酶(inducible nitric oxide synthase，iNOS)及促炎因子肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、白细胞介素-1β(interleukin-1β，IL-1β)、白细胞介素-6(interleukin-6，IL-6)的mRNA表达；M2型小胶质细胞/巨噬细胞表面标记物CD206、精氨酸酶-1(arginase-1，Arg-1)及抗炎因子白细胞介素-10(interleukin-10, IL-10)、转化生长因子-β(transforming growth factor -β，TGF-β)的mRNA表达。[结果]免疫荧光双标染色结果表明，与模型组比较，BYHWD显著减少缺血区M1型小胶质细胞/巨噬细胞(CD16/32+)数量(P＜0.01)，增加M2型小胶质细胞(CD206+)数量(P＜0.05)。qRT-PCR结果表明，与模型组比较，BYHWD显著下调M1型小胶质细胞/巨噬细胞表面标记物CD86、iNOS和促炎因子TNF-α、IL-1β、IL-6 mRNA表达(P＜0.01)，上调M2型小胶质细胞/巨噬细胞表面标记物CD206、Arg-1和抗炎因子IL-10、TGF-β mRNA表达(P＜0.01)。[结论] BYHWD可能通过促进激活的小胶质细胞/巨噬细胞从M1型向M2型转换，从而抑制大鼠脑缺血后炎症反应。 |
| 英文摘要: |
| [Objective] To investigate the effects of Buyang Huanwu Decoction(BYHWD) on microglia/macrophage M1/M2 polarization and neuro-inflammation after cerebral ischemia in rats. [Methods] Focal cerebral ischemia was induced by occlusion of the right middle cerebral artery for 90 minutes using the intraluminal filament model. The rats were randomly divided into sham operation group, model group and BYHWD group. BYHWD (13g·kg-1) was administered intragastricly in BYHWD group 24h after ischemia and once daily for 14 days. Microglia/macrophage M1/M2 phenotype was determined by double immune-fluorescence labeling. The expression of M1/M2 microglia/macrophage markers were detected by qRT-PCR. [Results] Compared with model group, double immune-fluorescence labeling showed that BYHWD significantly reduced the expression of M1 marker CD16/32(P＜0.01) and enhanced the expression of M2 marker CD206 in microglia/macrophage after cerebral ischemia(P＜0.05). qRT-PCR results showed that BYHWD significantly down-regulated the expression of M1 microglia/macrophage surface markers CD86, inducible nitric oxide synthase (iNOS), and pro-inflammatory cytokines tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6) mRNA(P＜0.01). In contrast, BYHWD up-regulated the expression of M2 microglia/macrophage surface markers CD206, arginase-1(Arg-1), and anti-inflammatory cytokines interleukin-10(IL-10) and transforming growth factor beta(TGF-β) mRNA(P＜0.01). [Conclusion] These findings suggest that BYHWD inhibits inflammation after cerebral ischemia in rats, which may be related to promoting the polarization of the microglia/macrophage from M1 to M2 phenotype in the ischemic brain. |
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