复方丹参滴丸抑制活性氧介导的内皮细胞损伤机制  

Mechanism of Compound Danshen Dripping Pills to Inhibit Endothelial Cell Damage Mediated by Reactive Oxygen Species

作　　者：郑丽丽 曹扬远 魏岚[1] ZHENG Lili;CAO Yangyuan;WEI Lan;LIU Meichen(Minxi vocational&Technical college;Pharmacy Department Fujian Medical Uniuersity Affliated Longyan First Hospital,Longyan,Fujian 364000,China)

机构地区：[1]闽西职业技术学院,福建龙岩364000

出　　处：《九江学院学报（自然科学版）》2024年第2期97-103,共7页Journal of Jiujiang University：Natural Science Edition

基　　金：福建省中青年教师教育科研项目(编号JAT210905);闽西职业技术学院科研项目(编号KJMY2310)的成果之一

摘　　要：目的探讨复方丹参滴丸抑制活性氧介导的内皮细胞损伤的机制。方法使用过氢化氢(H_(2)O_(2))构建内皮细胞损伤模型,将细胞分为正常组、模型组、复方丹参滴丸(50、100、150μg/mL)组,CCK-8检测细胞增殖情况,LDH试剂盒及ATP试剂盒检测细胞毒性及ATP产生,比色法检测内皮细胞MDA含量及SOD活力;ROS荧光探针检测细胞中ROS含量;Western blot及实时荧光定量PCR检测各组细胞中Bcl-2、Bax及Cleaved caspase-3的蛋白及mRNA表达情况。在上述模型的基础上使用ROS清除剂(NAC)构建ROS清除模型,将细胞分为正常组,H_(2)O_(2)组,H_(2)O_(2)+复方丹参100μg/mL组、H_(2)O_(2)+复方丹参100μg/mL+NAC组,免疫荧光探针、CCK-8、LDH及ATP试剂盒、比色法检测细胞ROS、增殖、毒性、ATP、MDA、SOD的含量及活力;Western blot及实时荧光定量PCR检测凋亡蛋白及mRNA表达情况。结果H_(2)O_(2)刺激后,细胞增殖被抑制,细胞毒性增强,ROS及MDA表达上调,ATP及SOD表达下调,Bcl-2表达下调,Bax及Cleaved caspase-3表达下调,复方丹参滴丸干预后可逆转内皮细胞损伤,以100μg/mL剂量组最显著。ROS清除剂抑制ROS的产生,并减弱100μg/mL复方丹参滴丸逆转内皮细胞损伤作用。结论复方丹参滴丸可逆转氧化应激产生的内皮细胞损伤,此作用可能是通过抑制ROS/Bcl-2/Bax信号通路实现的。Objective To explore the mechanism of compound Danshen dripping pills inhibiting endothelial cell damage mediated by reactive oxygen species.Method H_(2)O_(2)was used to construct endothelial cell injury model,and the cells were divided into normal group,model group,compound Danshen dripping pills(50,100,150μg/ml)groups.Cell proliferation was detected by CCK-8,and cytotoxicity and ATP production were detected by LDH and ATP kit.MDA content and SOD activity of endothelial cells were detected by colorimetry.ROS fluorescence probe was used to detect ROS content in cells.The protein and mRNA expression of Bcl-2,Bax and Cleaved caspase-3 were detected by Western blot and real-time quantitative PCR.On the basis of the above models,ROS scavenging model was constructed using ROS scavengers(NAC).The cells were divided into normal group,H_(2)O_(2)group,H_(2)O_(2)+compound Danshen 100μg/mL,H_(2)O_(2)+compound Danshen 100μg/mL+NAC group.The contents and activities of ROS,proliferation,toxicity,ATP,MDA and SOD were detected by immunofluorescence probe,CCK-8,LDH and ATP kit and colorimetry.The expressions of apoptotic proteins and mRNA were detected by Western blot and real-time quantitative PCR.Result After H_(2)O_(2)stimulation,cell proliferation was inhibited and cytotoxicity was enhanced,ROS and MDA expression was up-regulated,ATP and SOD expression was down-regulated,Bcl-2 expression was down-regulated,Bax and Cleaved caspase-3 expression were down-regulated,and endothelial cell damage was reversed after compound Danshen-based dropping pills,which was most significant in the 100μg/mL group.ROS scavenger inhibited ROS production and weakened the effect of 100μg/mL compound Danshen dripping pills on reversing endothelial cell damage.Conclusion Compound Danshen dripping pills can reverse endothelial cell damage caused by oxidative stress,which might be achieved by inhibiting ROS/Bcl-2/Bax signaling pathway.

关 键 词：复方丹参滴丸 活性氧 内皮细胞 增殖 凋亡 

分 类 号：R153.3[医药卫生—营养与食品卫生学]