对氧磷酶2对氧化低密度脂蛋白诱导的乳鼠心肌细胞损伤的影响  

Protective effect of paraoxonase 2 against oxidized low-density lipoprotein-induced neonatal rat myocardial cell injury in vitro

作　　者：霍金秀 马彦卓 王冬梅[2] 唐丽娜[2] 啜玉彩 昝晓花 陈瑜 

机构地区：[1]河北北方学院研究生部,河北张家口075000 [2]解放军白求恩国际和平医院心内科,石家庄050082 [3]解放军白求恩国际和平医院老年病科,石家庄050082

出　　处：《第三军医大学学报》2018年第2期130-135,共6页Journal of Third Military Medical University

基　　金：河北省重点基础研究项目(13967710D)~~

摘　　要：目的探讨对氧磷酶2(paraoxonase 2,PON2)在氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的乳鼠心肌细胞(neonatal rat ventricular cells,NRVCs)损伤中的作用及机制。方法分离SD大鼠(出生1~3 d,雌雄不限,共90只)心肌细胞。细胞分为:(1)对照组(未进行任何干预)、(2)0.1μmol/Lox-LDL组(ox-LDL干预24 h)、(3)10μmol/Lsalubrinal+ox-LDL组(salubrinal预处理1h后加入ox-LDL干预24 h)、(4)20μg/mLPON2+ox-LDL组(PON2预处理1 h后加入ox-LDL干预24 h)、(5)2.5μmol/LMn(Ⅲ)TBAP+ox-LDL组[(Mn(Ⅲ)TBAP预处理1 h后加入ox-LDL干预24 h]。通过MTT法和Caspase-3/7活性检测试剂盒分别检测细胞活性和细胞凋亡变化(n=6);Western blot检测心肌细胞内质网应激和氧化应激蛋白表达情况(n=4)。结果与对照组相比,ox-LDL明显降低心肌细胞活性[(0.417±0.047)vs(0.883±0.064),P<0.01]显著增加心肌细胞凋亡[(11 505±817)vs(6417±439),P<0.01]。与ox-LDL组相比,PON2、salubrinal和Mn(Ⅲ)TBAP干预后,细胞活性显著升高[(0.567±0.066)、0.649±0.082)、0.539±0.049)vs 0.417±0.047),P<0.01],细胞凋亡显著下降[(7 776±488)、(7 545±547)、(7 960±480)vs(11 505±817),P<0.01)。与对照组相比,ox-LDL显著增加PON2蛋白表达[(0.86±0.223)vs(0.561±0.13),P<0.05]、内质网应激蛋白eIF-2α^(phox)、caspase-12和氧化应激蛋白Nox2/gp91^(phox)、p47^(phox)表达[(1.309±0.274)vs(0.780±0.186)、(1.294±0.273)vs(0.606±0.064)、(1.342±0.274)vs(0.788±0.137)、(1.178±0.194)vs(0.629±0.125),P<0.01]。与ox-LDL组相比,PON2干预后,PON2蛋白表达明显升高[(1.190±0.151)vs(0.860±0.222),P<0.05];eIF-2α^(phox)、caspase-12、Nox2/gp91^(phox)和p47^(phox)蛋白表达明显降低[(0.924±0.170)vs(1.309±0.274)、(0.895±0.202)vs(1.294±0.273)、(0.957±0.190)vs(1.342±0.274)、(0.799±0.232)vs(1.178±0.194),P<0.05]。与PON2作用类似,salubrinal干预后,eIF-2α^(phox)和caspase-12蛋白表达明显降低[(1.010±0.096)vs(1.309±0.274)、(0.788±0.042)vs(1.294±0.273),P<0.01],Mn(ⅢObjectiveTo investigate the effects of paraoxonase 2 （PON2） in ameliorating neonatal rat ventricular cell （NRVC） injury induced by oxidized low density lipoprotein （oxLDL） and explore the underlying mechanism. MethodsNRVCs isolated from 90 neonatal （1~3 days old） SpragueDawley rats were exposed to 0.1 μmol/L oxLDL with or without pretreatment for 1 h with 10 μmol/L salubrinal, 20 μmol/L PON2, or 2.5 μmol/L Mn（III）TBAP. The cell viability and apoptosis were detected with MTT assay and caspase3/7 activity assay kit, and Western blotting was used to detect the expression of proteins related with endoplasmic reticulum stress （ERS） and oxidative stress （OS）. ResultsExposure to oxLDL resulted in significantly reduced cell viability （0.417±0.047 vs 0.883±0.064, P〈0.01） and increased number of apoptotic cells （11 505±817 vs 6 417±439, P〈0.01） in neonatal rat cardiac cells. Compared with cells exposed to oxLDL alone, the cells pretreated with PON2, salubrinal or Mn（III）TBAP exhibited significantly increased viability （0.567±0.066, 0.649±0.082, and 0.539±0.049, respectively; P〈0.01） and decreased number of apoptotic cells （7 776±488, 7 545±547, and 7 960±480, respectively; P〈0.01）. Compared with the control cells, exposure to oxLDL markedly increased the expression of PON2 in the cardiac cells （P〈0.05）, ERSrelated proteins eIF2α^phox（P〈0.01） and caspsae12 （P〈0.01）, and OSrelated proteins Nox2/gp91^phox （P〈0.01） and p47^phox （P〈0.01）; pretreatment of the cells with PON2 significantly decreased the protein expressions of eIF2α^phox caspase12, Nox2/gp91^phox, and p47^phox and enhanced PON2 expression （P〈0.05）; pretreatment with salubrinal significantly decreased the protein expressions of eIF2α^phoxand caspsae12 （P〈0.01）, Mn（Ⅲ）TBAP significantly decreased the protein expressions of Nox2/gp91^phox and p47^phox （P〈0.01）, and they both significantly increased

关 键 词：对氧磷酶2 氧化低密度脂蛋白 内质网应激 氧化应激 凋亡 

分 类 号：R-322[医药卫生] R542.2