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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 - 第三军医大学学报 2018年40卷第2期 页码：130-135
- 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：
[王冬梅，E-mail：slwangdm@126.com WANG Dongmei, E-mail: slwangdm@126.com
- 基金信息：河北省重点基础研究项目(13967710D)
DOI：10.16016/j.1000-5404.201708079
中图分类号： R-322;R542.2;R977.3收稿：2017-08-14，
修回：2017-11-02，
纸质出版：2018-01-30
- 稿件说明：
移动端阅览
霍金秀, 马彦卓, 王冬梅, 等. 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[J]. 第三军医大学学报, 2018,40(2):130-135.
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.霍金秀, 马彦卓, 王冬梅, 等. 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[J]. 第三军医大学学报, 2018,40(2):130-135. DOI： 10.16016/j.1000-5404.201708079.
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
2
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
2
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(
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
2
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，e
IF-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(Ⅲ)TBAP干预后，Nox2/gp91
phox
和p47
phox
蛋白表达明显降低[(0.898±0.190)
vs
(1.342±0.274)、(0.769±0.158)
vs
(1.178±0.194)，
P
<
0.01
]
，二者均显著升高PON2蛋白表达[(1.248±0.259)、(1.176±0.248)
vs
(0.860±0.222)，
P
<
0.05
]
。
结论
2
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
2
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
2
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
2
Exposure 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 wit
h 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
phox
and 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 the expression of PON2 (
P
<
0.05).
Conclusion
2
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
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