雷公藤甲素调节小胶质细胞极化反应保护损伤的神经元  

Triptolide protects damaged neurons by regulating microglial polarization

作　　者：张慧宇 于婧文[2] 白振军 李亮 穆秉桃[2] 张金锋 解佳伟 Zhang Huiyu;Yu Jingwen;Bai Zhenjun;Li Liang;Mu Bingtao;Zhang Jinfeng;Xie Jiawei(College of Traditional Chinese Medicine Health Service,Shanxi Datong University,Datong 037009,Shanxi Province,China;Medical College,Shanxi Datong University,Datong 037009,Shanxi Province,China)

机构地区：[1]山西大同大学中医药健康服务学院,山西省大同市037009 [2]山西大同大学医学院,山西省大同市037009

出　　处：《中国组织工程研究》2023年第33期5342-5347,共6页Chinese Journal of Tissue Engineering Research

基　　金：山西省基础研究计划项目(20210302123478),项目负责人:张慧宇;山西省基础研究计划项目(20210302123337),项目负责人:于婧文。

摘　　要：背景:将小胶质细胞从M1表型转变为M2表型被认为是治疗神经退行性疾病的一种有希望的策略。多项研究表明,雷公藤甲素可以抑制神经炎症,改善多种神经退行性疾病,但是其机制尚不明确。目的:探讨雷公藤甲素对脂多糖激活小胶质细胞诱导SH-SY5Y细胞损伤的保护作用及其机制。方法:CCK8法检测BV2细胞活性筛选最佳的雷公藤甲素使用浓度;然后将BV2细胞分为3组:对照组,模型组(1μg/mL脂多糖),雷公藤甲素组(1 nmol/L雷公藤甲素+1μg/mL脂多糖),收集上清液,采用Griess法检测一氧化氮水平,ELISA法检测促炎因子白细胞介素6、肿瘤坏死因子α、白细胞介素1β和抗炎因子白细胞介素10水平;免疫荧光染色和Western blot法检测BV2细胞诱导型一氧化氮合酶和精氨酸酶1的表达。收集3组小胶质细胞条件培养液并分别作用于SH-SY5Y细胞:分为对照条件培养液组、模型条件培养液组和雷公藤甲素条件培养液组,TUNEL染色法检测SH-SY5Y细胞凋亡率,免疫荧光染色和Western blot法检测caspase3、Bax和Bcl-2的表达。结果与结论:(1)与对照组比较,模型组BV2细胞上清液中一氧化氮、白细胞介素6、肿瘤坏死因子α和白细胞介素1β的释放量明显增加,白细胞介素10的释放量明显减少,诱导型一氧化氮合酶蛋白的表达增加,精氨酸酶1蛋白的表达减少;与模型组比较,雷公藤甲素组BV2细胞上清液中一氧化氮、白细胞介素6、肿瘤坏死因子α和白细胞介素1β水平降低,白细胞介素10水平增加,诱导型一氧化氮合酶蛋白的表达下降,精氨酸酶1蛋白的表达增加;(2)与对照条件培养液组比较,模型条件培养液组SH-SY5Y细胞的凋亡率增加,caspase3、Bax表达增加,Bcl-2表达减少;与模型条件培养液组比较,雷公藤甲素条件培养液组SH-SY5Y细胞凋亡率下降,caspase3、Bax表达减少,Bcl-2表达增加;(3)结果说明,雷公藤甲素可以减轻脂多糖激活小胶质细胞诱BACKGROUND:Transforming microglia from the M1 phenotype to the M2 phenotype is considered to be a promising strategy for the treatment of neurodegenerative diseases.Many studies have shown that triptolide can inhibit neuroinflammation and alleviate a variety of neurodegenerative diseases,but its mechanism is still unclear.OBJECTIVE:To investigate the protective effect of triptolide on SH-SY5Y cell injury induced by lipopolysaccharide-activated microglia and its mechanism.METHODS:CCK8 assay was used to detect the viability of BV2 cells and screen the best concentration of triptolide.The BV2 cells were divided into three groups:control group,model group(1μg/mL lipopolysaccharide),and triptolide group(1 nmol/L triptolide+1μg/mL lipopolysaccharide).The supernatant was collected and the content of nitric oxide was detected by Griess assay.The levels of proinflammatory cytokines interleukin 6,tumor necrosis factorα,interleukin-1βand anti-inflammatory factor interleukin 10 were measured by ELISA.The expression levels of inducible nitric oxide synthase and arginase 1 in BV2 cells were determined by immunofluorescence staining and western blot assay.Three groups of microglia-conditioned medium were collected and treated on SH-SY5Y cells respectively:control-microglia-conditioned medium group,model-microglia-conditioned medium group and triptolide-microglia-conditioned medium group.TUNEL staining was utilized to detect the cell apoptosis rate in SH-SY5Y cells.Immunofluorescence staining and western blot assay were used to examine the expression of caspase 3,Bax and Bcl-2.RESULTS AND CONCLUSION:(1)Compared with the control group,the levels of nitric oxide,interleukin-6,tumor necrosis factorαand interleukin-1βwere significantly increased,and the level of interleukin-10 was significantly decreased,and the expression of inducible nitric oxide synthase protein increased,the expression of arginase 1 protein decreased in the supernatant of BV2 cells in the model group.Compared with the model group,the levels of nitric oxide

关 键 词：雷公藤甲素 脂多糖 小胶质细胞 炎症因子 极化 神经元 凋亡 

分 类 号：R459.9[医药卫生—治疗学] R392.5[医药卫生—临床医学] R285.5