| 段 鹏,霍 康,王建懿,郭秦乐,李 娜,许 静.牡荆素对大鼠脑缺血再灌注损伤后的小胶质细胞/巨噬细胞M2极化的影响[J].,2022,(19):3618-3624 |
| 牡荆素对大鼠脑缺血再灌注损伤后的小胶质细胞/巨噬细胞M2极化的影响 |
| Effect of Vitexin on M2 Polarization of Microglia/Macrophages after Cerebral Ischemia-reperfusion Injury in Rats |
| 投稿时间：2022-04-22 修订日期：2022-05-17 |
| DOI：10.13241/j.cnki.pmb.2022.19.003 |
| 中文关键词: 牡荆素 脑缺血再灌注损伤 小胶质细胞/巨噬细胞 M2极化 TLR4/NF-κB信号通路 |
| 英文关键词: Vitexin Cerebral ischemia-reperfusion injury Microglia/macrophages M2 polarization TLR4/NF-κB signaling pathway |
| 基金项目:陕西省自然科学基础研究计划项目（2018JM7021）；西安交通大学第一附属医院基金自由探索项目（2019ZYTS-10） |
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| 中文摘要: |
| 摘要 目的：探究牡荆素（Vitexin）对大鼠脑缺血再灌注损伤（CIRI）后的小胶质细胞巨噬细胞M2极化的影响。方法：采用改良的Longa法建立大鼠右侧中动脉阻塞/再灌注（MCAO/R）模型。建模后，将大鼠分为假手术组（Sham，n=10）、MCAO/R组（n=12）、Vitexin组（n=12）和Vitexin+脂多糖（LPS）组（n=12）。Sham组和MCAO/R组腹腔注射2 mL 0.9%生理盐水，Vitexin组腹腔注射2 mL牡荆素溶液（3 mg/kg），Vitexin+LPS组腹腔注射1 mL牡荆素溶液（3 mg/kg）和1 mL LPS溶液（0.5 mg/kg）。1次/d，连续14 d。给药14 d后，根据Zea Longa 5分法对大鼠进行神经功能评分。通过2,3,5-三苯基四唑氯化物（TTC）法检测脑梗死体积。采用双重免疫荧光染色方法检测大脑缺血半影区CD16/32+/Iba1+、CD206+/Iba1+的共表达。通过qRT-PCR或Western blot检测大脑缺血半影区Toll样受体4（TLR4）、肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、IL-1β、IL-4、IL-10、核因子-κB（NF-κB）p65的mRNA或蛋白表达。使用商用试剂盒检测大脑缺血半影区丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）的含量。结果：与MCAO/R组相比，Vitexin组神经功能评分降低，脑梗死体积降低，MDA水平降低，SOD和GSH-Px水平升高（P<0.05）。与MCAO/R组相比，Vitexin组CD16/32+/Iba1+的阳性细胞数量降低，CD206+/Iba1+的阳性细胞数量升高，TNF-α和IL-1β的mRNA水平降低，而IL-4和IL-10的mRNA水平升高（P<0.05）。与MCAO/R组相比，Vitexin组TLR4 mRNA和蛋白水平降低，细胞核NF-κB p65的蛋白表达水平降低（P<0.05）。然而，LPS给药逆转了牡荆素对上述指标的影响（P<0.05）。结论：牡荆素部分通过TLR4/NF-κB信号通路促进大鼠CIRI后小胶质细胞/巨噬细胞向M2表型极化，从而促进神经功能恢复。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of Vitexin on the M2 polarization of microglia and macrophages after cerebral ischemia-reperfusion injury (CIRI) in rats. Methods: The modified Longa method was used to establish the right middle artery occlusion/reperfusion (MCAO/R) model in rats. After modeling, the rats were divided into sham group (Sham, n=10), MCAO/R group (n=12), Vitexin group (n=12) and Vitexin+lipopolysaccharide (LPS) group (n=12). The Sham group and MCAO/R group were injected intraperitoneally with 2 mL 0.9% saline, the Vitexin group was intraperitoneally injected with 2 mL vitexin solution (3 mg/kg), and the Vitexin+LPS group was intraperitoneally injected with 1 mL of vitexin solution (3 mg/kg) and 1 mL LPS solution (0.5 mg/kg). 1 time/d, continuous 14 d. After 14 days of administration, the rats were scored for neurological function according to the Zea Longa 5-point method. The volume of cerebral infarction was detected by 2,3,5-triphenyltetrazolium chloride (TTC) method. The double immunofluorescence staining method was used to detect the co-expression of CD16/32+/Iba1+ and CD206+/Iba1+ in the cerebral ischemic penumbra. The mRNA or protein expression of Toll-like receptor 4 (TLR4), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-1β, IL-4, IL-10, nuclear factor-κB (NF-κB) p65 in the cerebral ischemic penumbra area were detected by qRT-PCR or Western blot. Commercial kits were used to detect the levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione the penumbra of cerebral ischemia. Results: Compared with the MCAO/R group, the Vitexin group had lower neurological scores, lower cerebral infarction volume, lower MDA levels, and higher SOD and GSH-Px levels(P<0.05). Compared with the MCAO/R group, the number of CD16/32+/Iba1+ positive cells in the Vitexin group decreased, the number of CD206+/Iba1+ positive cells increased, the mRNA levels of TNF-α and IL-1β decreased, while IL -4 and IL-10 mRNA levels increased (P<0.05). Compared with the MCAO/R group, the TLR4 mRNA and protein levels in the Vitexin group decreased, and the nuclear NF-κB p65 protein expression level decreased (P<0.05). However, LPS administration reversed the effect of vitexin on the above indicators (P<0.05). Conclusion: Vitexin promotes the polarization of microglia/macrophages to the M2 phenotype after CIRI in rats partly through the TLR4/NF-κB signaling pathway, thereby promoting the recovery of nerve function. |
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