ST2825通过调节小胶质细胞极化减轻脑缺血-再灌注损伤的研究  

ST2825 alleviates cerebral ischemia-reperfusion injury by regulating microglia polarization

作　　者：王坤[1,2] 白洁[1] 孙兴元 WANG Kun;BAI Jie;SUN Xingyuan(Jiamusi University,Jiamusi 154000,China;The Second Affiliated Hospital of Qiqihar Medical University,Qiqihar 161000,China)

机构地区：[1]佳木斯大学,黑龙江佳木斯154000 [2]齐齐哈尔医学院附属第二医院,黑龙江齐齐哈尔161000

出　　处：《中国实用神经疾病杂志》2024年第6期767-773,共7页Chinese Journal of Practical Nervous Diseases

基　　金：齐齐哈尔市科技计划创新激励项目(编号:CSFGG-2020227)。

摘　　要：目的研究ST2825对脑缺血-再灌注损伤小胶质细胞的作用。方法处于对数生长期小鼠小胶质细胞BV2随机分为对照组(Con)、模型组(Mod)和ST2825低剂量处理组(ST2825L)、ST2825高剂量处理组(ST2825H),采用氧糖剥夺/再复氧(OGD/R)诱导缺血-再灌注模型,对细胞进行ST2825低剂量(5μmol/L)、高剂量(10μmol/L)处理。采用cell counting kit-8(CCK-8)方法检测活细胞数量,采用流式细胞术检测细胞活性氧(ROS)水平、凋亡及细胞极化状态,采用qPCR检测TNF-α、IL-10基因表达水平,采用免疫印迹(Western blotting)方法检测Caspase-1、cleaced Caspase-1、NLRP3、iNOS、IL-1及IL-18蛋白表达水平。结果CCK-8显示,与Con组相比,OGD/R后Mod组细胞活力极显著降低(P<0.01);与Mod组相比,ST2825L组、ST2825H组细胞活力显著升高;Mod组较Con组ROS水平增加,ST2825处理后ROS有所下降;与Con组相比,Mod组细胞凋亡率增加,ST2825处理后凋亡率较Mod降低,且呈剂量依赖性。与Con组相比,Mod组CD86表达显著增加,CD163表达降低,提示Mod组细胞向M1型细胞极化;与Mod组相比,两种剂量ST2825处理后CD86表达降低,CD163表达增加,提示ST2825促进缺氧复氧的BV2细胞向M2型细胞极化。qPCR显示,与Con组相比,Mod组细胞TNF-α表达增加,IL-10表达降低,ST2825L组和ST2825H组TNF-α表达相较于Mod组降低,IL-10升高。Western blotting显示,与Con相比,Mod组ARG1、Caspase-1和cleaced Caspase-1表达显著降低,NLRP3、iNOS、IL-1及IL-18表达增加,ST2825低剂量和高剂量处理后ARG1、Caspase-1和cleaced Caspase-1水平均显著增加,NLRP3、iNOS、IL-1及IL-18水平均降低。结论ST2825通过调节小胶质细胞极化减轻脑缺血-再灌注损伤,降低氧化应激及炎症反应,这是减轻脑缺血-再灌注损伤的新途径。Objective To analyze the effect of ST2825 on microglia following cerebral ischemiareperfusion injury.Methods Microglia BV2 of mice in logarithmic long-term were randomly divided into control group(Con),model group(Mod),ST2825 low-dose treatment group(ST2825L),ST2825 high-dose treatment group(ST2825H),and oxygen-glucose deprivation/re-oxygenation(OGD/R)was used to induce ischemia reperfusion model.Cells were treated with ST2825 low-dose(5μmol/L)and high dose(10μmol/L)processing.CCK-8 method was used to detect number of living cell,flow cytometry was employed to measure cell reactive oxygen species(ROS)levels,apoptosis,and cell polarization status,and qPCR was used to detect TNF-α,IL-10 genes expression levels.Western blotting and methods were used to detect the expression levels of Caspase-1,cleared Caspase-1,NLRP3,iNOS,IL-1,and IL-18 proteins.Results CCK-8 showed that after OGD/R,the cell viability of the Mod group was significantly reduced in comparison to the Con group(P<0.01).Compared with the Mod group,the cell viabilities of the ST2825L group and ST2825H group were significantly increased.The ROS level in the Mod group increased compared to the Con group,while ROS decreased after ST2825 treatment.In comparison to the Con group,the apoptosis rate in the Mod group increased,while the apoptosis rate in the ST2825 treatment decreased in a dose-dependent manner.When compared to the Con group,the Mod group showed a significant increase in CD86 expression and an increase in a decrease in CD163 expression,indicating that the Mod group cells were polarized towards M1 type cells.After treatment with two doses of ST2825,the Mod group showed a decrease in CD86 expression and an increase in CD163 expression,indicating that ST2825 promotes the polarization of hypoxic reoxygenated BV2 cells towards M2 type cells.qPCR showed that TNF-αin cells of the Mod group increased,IL-10 decreased compared to the Con group,and TNF-αin the ST2825L group and ST2825H group decreased compared to the Mod group,IL-10 increased.Western b

关 键 词：ST2825 小胶质细胞 脑缺血-再灌注损伤 氧化应激 炎症反应 细胞凋亡 细胞极化 小鼠 

分 类 号：R-332[医药卫生]