毛蕊花糖苷抑制NLRP3炎性小体活化改善BV-2细胞炎性损伤  

Verbascoside Inhibits the Activation of NLRP3 Inflammasome and Improves BV-2 Cell Inflammatory Damage

作　　者：黄文静 程芳 杜敏 陈佳艺 向维 石珂 谢兴亮[1] 盛艳梅[1] HUANG Wenjing;CHENG Fang;DU Min;CHEN Jiayi;XIANG Wei;SHI Ke;XIE Xingliang;SHENG Yanmei(Department of Pharmacy,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院药学院,四川成都610500

出　　处：《食品工业科技》2025年第4期367-373,共7页Science and Technology of Food Industry

基　　金：发育与再生四川省重点实验室开放基金(24FYYZS08);四川省大学生创新创业项目(S202313705044)。

摘　　要：目的:探讨桂花中的主要成分毛蕊花糖苷(Verbascoside,VB)抑制NLRP3炎性小体活化改善BV-2细胞炎性损伤及相关机制。方法:利用糖氧剥夺(Oxygen and glucose deprivation,OGD)法诱导BV-2细胞建立炎性损伤模型,实验分为正常对照组、OGD模型组、桂花乙醇提取物(Osmanthus fragrans ethanol extract,OEE)处理组(OGD+OEE)、VB处理组(OGD+VB)。采用MTT法检测BV-2细胞活力;使用乳酸脱氢酶(Lactate dehydrogenase,LDH)细胞毒性试剂盒检测培养液上清中LDH释放量,评价其对细胞膜的影响;利用Hoechst33342/PI双染技术检测其对细胞焦亡的影响;此外,采用免疫印迹(Western blot)法研究VB对NLRP3、Caspase-1、IL-1β及TNF-α等炎症相关蛋白的表达水平。结果:MTT法结果表明,OEE和VB均能显著促进BV-2细胞存活(P<0.001);LDH检测结果显示,OEE和VB均能极显著地减少培养液上清中LDH的释放(P<0.001),可减少细胞膜损伤;采用Hoechst 33342/PI双染后,OEE和VB组中BV-2细胞的PI阳性细胞数量均明显减少(P<0.01),荧光强度明显减弱。Western blot结果显示,经VB干预后,NLRP3/Caspase-1/IL-1β信号通路细胞焦亡蛋白水平显著降低(P<0.05或P<0.01)。结论:VB能够改善OGD诱导的BV-2细胞炎性损伤,可能是OEE抗炎的主要药效物质基础,其机制与抑制NLRP3/Caspase-1/IL-1β信号通路相关。Objective:To investigate the main component verbascoside(VB)in Osmanthus fragrans which inhibits NLRP3 inflammasome activation to improve inflammatory damage in BV-2 cells and its related mechanisms.Methods:A model of inflammatory damage induced by oxygen-glucose deprivation(OGD)in BV-2 cells was employed.The experiment was divided into 4 groups:The normal control group,the OGD model group,the Osmanthus fragrans ethanol extract group(OGD+OEE),and VB group(OGD+VB).The MTT assay was used to evaluate the effects of the treatments on BV-2 cell viability.The release of lactate dehydrogenase(LDH)in the culture supernatant was measured using an LDH cytotoxicity assay kit to assess the impact on the cell membrane.Hoechst 33342/PI double staining was utilized to examine the effect of the treatments on cell pyroptosis.Furthermore,Western blot analysis was conducted to investigate the expression levels of proteins related to the NLRP3 inflammasome pathway,including NLRP3,Caspase-1,IL-1β,and TNF-α.Results:The MTT assay results indicated that both OEE and VB significantly promoted the viability of BV-2 cells(P<0.001).LDH assay results demonstrated that both OEE and VB markedly reduced the release of LDH in the culture supernatant(P<0.001),indicating reduced damage to the cell membrane.Hoechst 33342/PI double staining revealed a significant decrease in the number of PI-positive cells in both the OEE and VB groups(P<0.01),with a marked reduction in fluorescence intensity.The Western blot results indicated that,after treatment with verbascoside,there was a significant decrease in the levels of cell pyroptosis proteins associated with the NLRP3/Caspase-1/IL-1βsignaling pathway(P<0.05 or P<0.01).Conclusion:VB can ameliorate OGD-induced inflammatory damage in BV-2 cells,which may constitute the primary pharmacodynamic basis for the anti-inflammatory effects of OEE.The mechanism is associated with the inhibition of the NLRP3/Caspase-1/IL-1βsignaling pathway.

关 键 词：毛蕊花糖苷 桂花 BV-2细胞 NLRP3炎性小体 OGD模型 

分 类 号：R285.5[医药卫生—中药学]