雷公藤红素抑制LDL及HAEC细胞氧化损伤作用  

The inhibitive effects of celastrol on LDL oxidation and HAEC cell oxidative damage

作　　者：李锋[1] 李义嘉 李清仙 郭养浩[1] 

机构地区：[1]福州大学药物生物技术与工程研究所,福建福州350116

出　　处：《中国药理学通报》2016年第11期1578-1584,共7页Chinese Pharmacological Bulletin

基　　金：国家海洋公益性行业科研专项(No 201205022)

摘　　要：目的考察雷公藤红素(celastrol)体外对低密度脂蛋白(LDL)氧化过程及动脉血管内皮细胞氧化损伤的抑制作用。方法采用体外Cu^(2+)诱导LDL氧化修饰的体外化学反应模型,设置阴性对照、模型对照及雷公藤各剂量组,测定Cu^(2+)诱导氧化LDL的动力学曲线,分析比较各实验组的曲线下面积(AUC)、延滞期持续时间(lag time)及脂质过氧化物产生量(TBARS值);MTT法测定细胞活性,以确定雷公藤红素对正常细胞的安全剂量;建立AAPH诱导的人主动脉内皮细胞HAEC氧化损伤的细胞模型,设置阴性对照、模型对照及雷公藤各剂量组,测定细胞的乳酸脱氢酶(LDH)漏出量、超氧化物岐化酶(SOD)及谷胱甘肽过氧化物酶(GPX)活性,Hoechst 33258荧光染色观察细胞核受损伤情况,RTq PCR法分析Nrf2、HO-1 mRNA表达水平。结果体外实验结果显示,celastrol在体外较低的剂量条件下(100 nmol·L^(-1)~1μmol·L^(-1))即能有效延长LDL氧化过程中的迟滞期,降低反应动力学曲线的AUC以及脂质过氧化物的生成,表明celastrol在体外能有效抑制Cu^(2+)诱导的LDL氧化。细胞实验结果显示,在100~400 nmol·L^(-1)剂量范围内,celastrol能有效降低AAPH所致HAEC细胞的LDH漏出量,维护细胞膜和细胞核的完整性;提高受损细胞的Nrf2和HO-1mRNA表达,提高受损细胞的SOD、GPX酶活,从而提升内皮细胞的抵抗氧化应激的能力。结论 celastrol可在体外有效抑制Cu^(2+)诱导人LDL氧化损伤,在无细胞毒安全剂量下(100~400 nmol·L^(-1))可抑制AAPH所致的HAEC细胞氧化应激损伤。Aim To evaluate the inhibitive effects of celastrol on LDL oxidation and HAEC cell oxidative damage. Methods The Cu^(2+)-induced LDL oxidation model was employed to evaluate celastrol inhibitive effect on LDL oxidation in vitro,the oxidative reaction kinetic curves were determined, and the AUC,lag time,TBARS value were assayed. The AAPH-induced HAEC damaging cellular model was employed to evaluate the effect of celastrol on oxidative cellular damage.The safe dose of celastrol on normal cells was determined by MTT method,and the effects of celastrol on HAEC oxidative damage were evaluated at the range of this safe dose. The LDH leakage,ROS level,SOD and GPX enzymatic activity,Nrf2 and HO-1 mRNA expression were determined. Results At the dose range of100 nmol· L^(-1)to 1 μmol · L^(-1),celastrol effectively extended the lag time of LDL oxidative process induced by Cu^(2+),reduced the AUC of oxidative reaction kinetic curve and reduced the generation of lipid peroxide in the LDL oxidative process. In the cellular experiment,celastrol effectively reduced the LDH leakage induced by AAPH,increased the integrity of cell membrane and nucleus,enhanced the antioxidative enzyme activities of cellular SOD,GPX and increased the expressions of Nrf2,HO-1 mRNA,celastrol also maintained the integrity of cellular structure. Conlusion Celastrol can effectively inhibit LDL oxidation induced by Cu^(2+),and can inhibit HAEC cell oxidative damage induced by AAPH at the dose range of 100 to 400 nmol·L^(-1).

关 键 词：雷公藤红素 低密度脂蛋白 血管内皮细胞 脂质过氧化 氧化应激 抗氧化 活性氧 

分 类 号：R284.1[医药卫生—中药学] R322.1[医药卫生—中医学]