Protective effect of paraoxonase 2 against oxidized low-density lipoprotein-induced neonatal rat myocardial cell injury <italic style="font-style: italic">in vitro</italic>

Jinxiu HUO; Yanzhuo MA; Dongmei WANG; Lina TAGN; Yucai CHUAI; Xiaohua ZAN; Yu CHEN

doi:10.16016/j.1000-5404.201708079

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

Basic Medicine | 更新时间：2026-01-09

- Protective effect of paraoxonase 2 against oxidized low-density lipoprotein-induced neonatal rat myocardial cell injury in vitro
- Journal of Third Military Medical University Vol. 40, Issue 2, Pages: 130-135(2018)
- 作者机构：
  
  1.075000 河北张家口，河北北方学院研究生部
  2.050082 石家庄，解放军白求恩国际和平医院：心内科
  3.050082 石家庄，解放军白求恩国际和平医院老年病科
- 作者简介：
- 基金信息：
  
  the Key Basic Research Project of Hebei Province(13967710D)
- DOI：10.16016/j.1000-5404.201708079
  CLC： R-322;R542.2;R977.3
- Received：14 August 2017，
  
  Revised：2017-11-02，
  
  Published：30 January 2018
- 稿件说明：
移动端阅览
Jinxiu HUO, Yanzhuo MA, Dongmei WANG, et al. Protective effect of paraoxonase 2 against oxidized low-density lipoprotein-induced neonatal rat myocardial cell injury in vitro[J]. Journal of Third Military Medical University, 2018, 40(2): 130-135.
DOI：

Jinxiu HUO, Yanzhuo MA, Dongmei WANG, et al. Protective effect of paraoxonase 2 against oxidized low-density lipoprotein-induced neonatal rat myocardial cell injury in vitro[J]. Journal of Third Military Medical University, 2018, 40(2): 130-135. DOI： 10.16016/j.1000-5404.201708079.

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)、②0.1 μmol/Lox-LDL组(ox-LDL干预24 h)、③10 μmol/Lsalubrinal+ ox-LDL组(salubrinal预处理1 h后加入ox-LDL干预24 h)、④20 μg/mLPON2+ ox-LDL组(PON2预处理1 h后加入ox-LDL干预24 h)、⑤2.5 μmol/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(

=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(

=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)

(0.883±0.064)，

0.01

]

显著增加心肌细胞凋亡[(11 505±817)

(6417±439)，

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)

0.417±0.047)，

0.01

]

，细胞凋亡显著下降[(7 776±488)、(7 545±547)、(7 960±480)

(11 505±817)，

0.01)。与对照组相比，ox-LDL显著增加PON2蛋白表达[(0.86±0.223)

(0.561±0.13)，

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)

(0.780±0.186)、(1.294±0.273)

(0.606±0.064)、(1.342±0.274)

(0.788±0.137)、(1.178±0.194)

(0.629±0.125)，

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)

(0.860±0.222)，

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)

(1.309±0.274)、(0.895±0.202)

(1.294±0.273)、(0.957±0.190)

(1.342±0.274)、(0.799±0.232)

(1.178±0.194)，

0.05

]

。与PON2作用类似，salubrinal干预后，e

IF-2α

phox

和caspase-12蛋白表达明显降低[(1.010±0.096)

(1.309±0.274)、(0.788±0.042)

(1.294±0.273)，

0.01

]

，Mn(Ⅲ)TBAP干预后，Nox2/gp91

phox

和p47

phox

蛋白表达明显降低[(0.898±0.190)

(1.342±0.274)、(0.769±0.158)

(1.178±0.194)，

0.01

]

，二者均显著升高PON2蛋白表达[(1.248±0.259)、(1.176±0.248)

(0.860±0.222)，

0.05

]

。

结论

PON2可能通过代偿性升高抑制氧化应激和内质网应激而减轻ox-LDL诱导的心肌细胞损伤。

Abstract

Objective

To 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.

Methods

NRVCs 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).

Results

Exposure to ox-LDL resulted in significantly reduced cell viability (0.417±0.047

0.883±0.064

0.01) and increased number of apoptotic cells (11 505±817

6 417±439

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;

0.01) and decreased number of apoptotic cells (7 776±488

7 545±547

and 7 960±480

respectively;

0.01). Compared wit

h the control cells

exposure to ox-LDL markedly increased the expression of PON2 in the cardiac cells (

0.05)

ERS-related proteins eIF-2α

phox

(

0.01) and caspsae-12 (

0.01)

and OS-related proteins Nox2/gp91

phox

(

0.01) and p47

phox

(

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 (

0.05); pretreatment with salubrinal significantly decreased the protein expressions of eIF-2α

phox

and caspsae-12 (

0.01)

Mn(Ⅲ)TBAP significantly decreased the protein expressions of Nox2/gp91

phox

and p47

phox

(

0.01)

and they both significantly increased the expression of PON2 (

0.05).

Conclusion

PON2 may provide cardioprotection against NRVC injury induced by ox-LDL by the compensatory elevation of PON2 to inhibit oxidative stress and endoplasmic reticulum stress.

关键词

Keywords

references

PARK C S, CHA H, KWON E J, et al .The chemical chaperone 4-phenylbutyric acid attenuates pressure-overload cardiac hypertrophy by alleviating endoplasmic reticulum stress[J ] .Biochem Biophys Res Commun, 2012, 421(3): 578-584.DOI: 10.1016/j.bbrc.2012.04.048.

HORKE S, WITTE I, WILGENBUS P, et al .Paraoxonase-2 reduces oxidative stress in vascular cells and decreases endoplasmic reticulum stress-induced caspase activation[J ] .Circulation, 2007, 115(15): 2055-2064.DOI: 10.1161/CIRCULATIONAHA.106.681700.

GAO S, LIU J.Association between circulating oxidized low-density lipoprotein and atherosclerotic cardiovascular disease[J].Chronic Dis Transl Med, 2017, 3(2): 89-94.DOI:10.1016/j.cdtm.2017.02.008.

CAMONT L, LHOMME M, RACHED F, et al .Small, dense high-density lipoprotein-3 particles are enriched in negatively charged phospholipids: relevance to cellular cholesterol efflux, antioxidative, antithrombotic, anti-inflammatory, and antiapoptotic functionalities[J ] .Arterioscler Thromb Vasc Biol.2013, 33(12): 2715-2723.DOI: 10.1161/ATVBAHA.113.301468.

TRPKOVIC A, RESANOVIC I, STANIMIROVIC J, et al .Oxidized low-density lipoprotein as a biomarker of cardiovascular diseases[J ] .Crit Rev Clin Lab Sci, 2015, 52(2): 70-85.DOI: 10.3109/10408363.2014.992063.

WU C Y, TANG Z H, JIANG L, et al .PCSK9 siRNA inhibits HUVEC apoptosis induced by ox-LDL via Bcl/Bax-caspase9-caspase3 pathway[J ] .Mol Biochem, 2012, 359(1-2): 347-358.DOI: 10.1007/s 11010-011-1028-6

LI W, CHEN Z, YAN M, et al .The protective role of isorhamnetin on human brain microvascular endothelial cells from cytotoxicity induced by methylglyoxal and oxygen-glucose deprivation[J ] .J Neurochem, 2015, 136(3): 651-659.DOI: 10.1111/jnc.13436.

LI H, HORKE S, FORSTERMANN U, et al .Vascular oxidative stress, nitric oxide and atherosclerosis[J ] .Atherosclerosis, 2014, 237(1): 208-219.DOI: 10.1016/j.athrosclerosis.2014.09.001.

YANG Y, CUEVAS S, YANG S, et al .Sestrin2 decreases renal oxidative stress, lowers blood pressure, and mediates dopamine D2 receptor-induced inhibition of reactive oxygen species production[J ] .Hypertension, 2014, 64(4): 825-32.DOI: 10.1161/HYPERTENSIONAHA.114.03840

AVIRAM M, ROSENBLAT M.Paraoxonases1, 2, and 3, oxidative stress, and macrophage foam cell formation during atherosclerosis development[J].Free Radic Biol Med.2004, 37(9): 1304-1316.DOI: 10.1016/j.freeradbiomed.2004.06.030

ROSENBLAT M, VOLKOVA N, PALAND N, et al.Triglyceride accumulation in macrophages upregulates paraoxonase 2(PON2) expression via ROS-mediated JNK/c-Jun signaling pathway activation[J].Biofactors, 2012, 38(6): 458-469.DOI: 10.1002/biof.1052.

QI JC, LIU P G, WANG C, et al .Tacrolimus protects vascular endothelial cells from injuries caused by Ox-LDL by regulating endoplasmic reticulum stress[J ] .Eur Rev Med Pharmacol Sci, 2017, 21(17): 3966-3973.

BAI S, CHENG L, YANG Y, et al .c1q/TNF-related protein 9 protects diabetic rat heart against ischemia reperfusion injury: role of endoplasmic reticulum stress[J ] .Oxid Med Cell Longev.2016, 2016: 1902025.DOI: 10.1155/2016/1902025.

张国强. PERK/eIF2α通路介导氧化型低密度脂蛋白诱导的内皮细胞凋亡及辛伐他汀的干预作用[D]. 长沙: 中南大学, 2012.

ZHANG G Q. Role of PERK/eIF2α in ox-LDL-induced endothelial cell apoptosis and the effect of Simvastatin[D]. Changsha: Central South University, 2012.

XU X C, LIU T T, ZHANG A M, et al .Reactive oxygen species-triggered trophoblast apoptosis is initiated by endoplasmic reticulum stress via activation of caspase-12, CHOP, and the JNK pathway in toxoplasma gondii infection in mice[J ] .Infection immune, 2012, 80(6): 2121-2132.DOI: 10.1128/IAI.06295-11.

WITTE I, ALTENHOFER S, WILQENBUS P, et al .Beyond reduction of atherosclerosis: PON2 provides apoptosis resistance and stabilizes tumor cells[J ] .Cell Death Dis, 2011, 13;2: e112.DOI: 10.1038/CDDIS.2010.91.

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

DOI：10.16016/j.1000-5404.201708079

摘要

Abstract

关键词

Keywords

references