α-细辛醚通过调控NLRP3通路保护氧糖剥夺/再灌注损伤BV2细胞的研究  

Study ofα-Asarone protecting BV2 cells damaged by OGD/R by regulating of NLRP3 pathway

作　　者：徐飞飞 崔恺 王立有 管雅琪 刘铭 李钦青 田雅娟 楚世峰[2] 贺文彬 XU Fei-fei;CUI Kai;WANG Li-you;GUAN Ya-qi;LIU Ming;LI Qin-qing;TIAN Ya-juan;CHU Shi-feng;HE Wen-bin(Shanxi Key Laboratory of Chinese Medicine Encephalopathy,Shanxi University of Chinese Medicine,Taiyuan 030619,China;Institute of Materia Medica,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050,China)

机构地区：[1]山西中医药大学,中医脑病学山西省重点实验室,山西太原030619 [2]中国医学科学院药物研究所,北京100050

出　　处：《中国药理学通报》2022年第8期1209-1218,共10页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No.81473375);山西省重点研发计划(国际科技合作)重大区域创新合作项目(No.201803D421006,201903D421018)。

摘　　要：目的评价α-细辛醚对脑缺血/再灌注损伤的小胶质细胞的保护作用及机制。方法采用α-细辛醚预处理BV2细胞,建立OGD/R模型模拟脑缺血/再灌注损伤。以CCK-8方法筛选α-细辛醚给药剂量及OGD/R模型的建立方法;观察细胞形态变化评价α-细辛醚对损伤细胞的影响;酶联免疫吸附法检测IL-1β、IL-18、IL-10、IL-4及ROS的水平;Western blot检测TNF-α、TGF-β以及NLRP3、caspase 1、NF-κB的表达量。结果α-细辛醚预处理能够明显降低OGD/R损伤细胞IL-1β和IL-18的释放,降低TNF-α的蛋白表达;能够明显升高IL-10和IL-4的释放,促进TGF-β蛋白表达,并呈量效关系,以α-细辛醚高剂量(16μmol·L^(-1))效果最佳。α-细辛醚还能够明显降低损伤细胞NLRP3、caspase 1、p-NF-κB的蛋白表达量及ROS的活性。结论α-细辛醚对脑缺血/再灌注损伤有明显的保护作用,主要是通过调控ROS活性,抑制NF-κB的磷酸化,以降低NLRP3炎症小体的过度活化,进而通过抗炎起到保护脑缺血/再灌注损伤的作用。Aim To evaluate the protective effect ofα-asarone on microglials with cerebral ischemia/reperfusion injury by measuring the expression of polar transformation and related inflammatory proteins in BV2 cells in vitro and its mechanisms.Methods The cerebral ischemia/reperfusion injury BV2 cells were pretreated byα-asarone in vitro and simulated by OGD/R model.The effect ofα-asarone on the viability of damaged cells in OGD/R model was determined by CCK-8;the morphological changes of cells were observed to analyze the general morphology of cells;the levels of proinflammatory factor IL-1β,IL-18 and anti-inflammatory factor IL-10,IL-4,and ROS activity secreted by BV2 cells were detected by ELISA;the protein expressions of TGF-β,TNF-αand inflammatory related protein NLRP3,caspase 1,p-NF-κB were detected by Western blot.Results The results of in vitro experiments were as follows:the activity of damaged cells in OGD/R model was significantly increased byα-asarone,with the increase of administration dose,the cells in the low,medium and high dose groups ofα-asarone decreased,and the“amoeba-like”cells and the cell body were gradually became stereoscopic and full.From the results of cell morphology,it could be seen thatα-asarone had a certain proliferative effect on normal cells;the release was significantly reduced of proinflammatory factor IL-1β,IL-18 and TNF-αin OGD/R injured BV2 cells pretreated withα-asarone,also increased the release of IL-10,IL-4 and TGF-β,with a dose-effect relationship,and the high dose(16μmol·L^(-1))was the best;the expressions of inflammatory related protein NLRP3,caspase 1,NF-κB and ROS activity in injured cells of OGD/R model were significantly reduced after pretreatment withα-asarone.Conclusionsα-asarone has a significant protective effect on cerebral ischemia/reperfusion injury,mainly by regulating ROS activity and inhibiting phosphorylation of NF-κB,in order to reduce the excessive activation of NLRP3 inflammatory corpuscles reducing the secretion of proinflammatory fact

关 键 词：Α-细辛醚 脑缺血/再灌注损伤 小胶质细胞 炎性反应 NLRP3 NF-κB 

分 类 号：R-332[医药卫生] R322.8R364.5R392.12R743.3R845.22R852.15