山麦冬皂苷B调节DAMPs释放诱导免疫源性细胞死亡的抗肿瘤机制研究  

Antitumor Effect of Spicatoside B Based on Regulating DAMPs Release to Induce Immunogenic Cell Death

作　　者：臧海洋 杨君伶[2] 陈秋峰 曹培 杨晓慧 ZANG Haiyang;YANG Junling;CHEN Qiufeng;CAO Pei;YANG Xiaohui(Nantong Hospital of Traditional Chinese Medicine,Nantong 226001,Jiangsu,China;Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu,China)

机构地区：[1]南通市中医院,江苏南通226001 [2]南通大学附属医院,江苏南通226001

出　　处：《辽宁中医杂志》2024年第11期150-153,I0002-I0004,共7页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82004279);南通市第六期江海英才培养项目(通委人才办[2022]12号);南通市青年医学重点人才项目(通卫科教[2021]15号)。

摘　　要：目的探讨Spicatoside B对肺癌细胞的增殖、凋亡和DAMPs释放的影响,并明确凋亡与DAMPs释放之间的关系。方法将不同浓度的SP-B作用于A549和H1299细胞株24 h,CCK8检测细胞增殖活性;根据CCK8结果,选取药物浓度,干预细胞24 h,流式细胞仪检测细胞凋亡;ATP试剂盒检测ATP细胞外分泌;流式细胞仪检测CRT表达;ELISA检测HMGB1、HSP70、HSP90的细胞外分泌;凋亡抑制剂抑制凋亡后,再次检测CRT、ATP、HMGB1和HSP70/90的表达。结果SP-B显著抑制A549和H1299细胞的增殖活性,IC_(50)值分别为73.11μM和51.461μM;选取40μM浓度干预细胞,可显著促进凋亡;药物作用后显著提高了CRT的阳性比率,增加了ATP、HMGB1、HSP70、HSP90的细胞外释放。抑制凋亡后,CRT、ATP、HMGB1和HSP70/90的表达随之减少。结论Spicatoside B可抑制增殖,促凋亡,并显著促进DAMPs的释放,诱导免疫原性细胞死亡,其中凋亡对DAMPs的释放发挥着重要作用。Objective To investigate the effects of Spicatoside B on the proliferation,apoptosis and DAMPs release of lung cancer cells and to clarify the relationship between apoptosis and DAMPs release.Methods Different concentrations of SP-B were applied to A549 and H1299 cell lines for 24 hours,and the cell proliferation activity was detected by CCK8.According to the results of CCK8,the drug concentration was selected,the cells were intervened for 24 hours,and the cell apoptosis was detected by flow cytometry.ATP extracellular secretion was detected by ATP kit.CRT expression was detected by flow cytometry.The extracellular release of HMGB1,HSP70 and HSP90 was detected by ELISA.After the apoptosis was inhibited,the expressions of CRT,ATP,HMGB1 and HSP70/90 were detected again.Results SP-B significantly inhibited the proliferation activity of A549 and H1299 cells and the IC_(50)values were 73.11μM and 51.461μM;40μM concentration of the drug can significantly promote the apoptosis,significantly increased the positive ratio of CRT and increased the extracellular release of ATP,HMGB1,HSP70 and HSP90.After inhibiting the apoptosis,the expressions of CRT,ATP,HMGB1 and HSP70/90 were decreased.Conclusion Spicatoside B can inhibit the proliferation,promote the apoptosis and significantly promote the release of DAMPs to induce immunogenic cell death.Apoptosis plays an important role in the release of DAMPs.

关 键 词：肺癌 Spicatoside B 凋亡 DAMPS ICD 

分 类 号：R256.13[医药卫生—中医内科学]