Carabin过表达对人心室肌细胞AC16氧糖剥夺后凋亡的影响

吴庆; 姚浩旗; 程阔菊; 唐建芳; 黄河; 陈建; 李洪; 杨天德

doi:10.16016/j.1000-5404.201807146

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Carabin过表达对人心室肌细胞AC16氧糖剥夺后凋亡的影响

基础医学 | 更新时间：2026-01-09

- Carabin过表达对人心室肌细胞AC16氧糖剥夺后凋亡的影响
- Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes
- 第三军医大学学报 2018年40卷第22期页码：2068-2073
- 作者机构：
  
  1.400037 重庆，陆军军医大学(第三军医大学)第二附属医院麻醉科
  2.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系
- 作者简介：
  
  [ 杨天德，电话:(023)68763638，E-mail：31011@sina.com YANG Tiande, Tel: 86-23-68763638, E-mail: 31011@sina.com
- 基金信息：
  
  国家自然科学基金面上项目(81370210)
- DOI：10.16016/j.1000-5404.201807146
  中图分类号： R322.11;R394-33;R394.2
- 收稿：2018-07-28，
  
  修回：2018-09-28，
  
  纸质出版：2018-11-30
- 稿件说明：
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吴庆, 姚浩旗, 程阔菊, 等. Carabin过表达对人心室肌细胞AC16氧糖剥夺后凋亡的影响[J]. 第三军医大学学报, 2018,40(22):2068-2073.

Qing WU, Haoqi YAO, Kuoju CHENG, et al. Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes[J]. Journal of Third Military Medical University, 2018, 40(22): 2068-2073.
吴庆, 姚浩旗, 程阔菊, 等. Carabin过表达对人心室肌细胞AC16氧糖剥夺后凋亡的影响[J]. 第三军医大学学报, 2018,40(22):2068-2073. DOI： 10.16016/j.1000-5404.201807146.

Qing WU, Haoqi YAO, Kuoju CHENG, et al. Effect of Carabin overexpression on oxygenglucose deprivation-induced apoptosis in cultured human ventricular myocytes[J]. Journal of Third Military Medical University, 2018, 40(22): 2068-2073. DOI： 10.16016/j.1000-5404.201807146.

摘要

目的

探讨Carabin基因过表达对氧糖剥夺(oxygen glucose deprivation

OGD)诱导的人心室肌细胞株AC16凋亡的作用。

方法

构建Carabin过表达及空载稳定细胞株，将AC16细胞分为空载常氧组、Carabin过表达常氧组、空载OGD 24 h组和Carabin过表达OGD 24 h组。空载常氧组及Carabin过表达常氧组置于正常条件下培养；空载OGD 24 h组和Carabin过表达OGD 24 h组置于Hanks液中放入缺氧恒温细胞培养箱培养24 h(1%O

-5%CO

-94%N

)。采用倒置显微镜观察细胞形态变化；流式细胞术检测心肌细胞凋亡率及存活率；TUNEL染色检测细胞凋亡情况；通过酶标仪检测心肌细胞凋亡蛋白酶Caspase-3和心肌细胞损伤标志物LDH的活性；Western blot检测细胞凋亡蛋白Caspase-3的表达水平。

结果

AC16细胞OGD 24 h后，细胞内Carabin的mRNA及蛋白水平显著降低。与空载常氧组比较，空载OGD 24 h组及Carabin过表达OGD 24 h组的细胞凋亡率、Caspase-3活性、LDH活性及Caspase-3蛋白表达水平均显著升高(

0.05)。与空载OGD 24 h组比较，Carabin过表达OGD 24 h组的细胞凋亡率、Caspase-3活性、LDH活性及Caspase-3蛋白表达水平显著降低(

0.05)。

结论

Carabin过表达能显著减轻OGD诱导后AC16细胞的凋亡。

Abstract

Objective

To investigate the effect of Carabin overexpression on apoptosis of human ventricular myocytes induced by oxygenglucose deprivation (OGD)

in vitro

Methods

Human ventricular myocyte AC16 cell line was transfected with a lentiviral vector carrying Carabin gene to induce Carabin overexpression or with a negative control vector. The transfected cells were cultured for 24 h under routine conditions or under OGD condition

s (in Hank's solution in a hypoxia incubator with 1% O

5% CO

and 94% N

). The changes in the morphology of AC16 cells were observed under an inverted microscope

and flow cytometry was used to determine the apoptosis rate and survival rate of the cells. The apoptotic cells were detected with TUNEL staining

and the activity of Caspase-3 and LDH in the cells were evaluated with enzyme labeling. Western blotting was performed to determine the protein expression of Caspase-3 in the cells in different culture conditions.

Results

OGD for 24 h significantly lowered the mRNA and protein levels of Carabin in AC16 cells. In the cells transfected with Carabin-overexpressing vector and the negative control vector

OGD for 24 h resulted in significantly increased cell apoptosis rate

Caspase-3 activity

LDH activity

and Caspase-3 protein expression as compared with the routinely cultured cells transfected with the negative control vector (

0.05). Compared with the negative control cells

the cells overexpressing Carabin were more resistant to OGD and showed significantly lower apoptotic rate

Caspase-3 and LDH activities

and Caspase-3 protein expression after OGD for 24 h (

0.05).

Conclusion

The overexpression of Carabin can significantly reduce the apoptosis of AC16 cells induced by OGD.

关键词

Keywords

references

YONEZU K, SAKAKURA K, WATANABE Y, et al . Determinants of survival and favorable neurologic outcomes in ischemic heart disease treated by veno-arterial extracorporeal m embrane oxygenation[J ] . Heart Vessels, 2018, 33(1): 25-32. DOI: 10.1007/s00380-017-1031-2.

ROMERO-CORRAL A, MONTORI V M, SOMERS V K, et al . Association of bodyweight with total mortality and with cardiovascular events in coronary artery disease: a systematic review of cohort studies[J ] . Lancet, 2006, 368(9536): 666-678. DOI: 10.1016/S0140-6736(06)69251-9.

PAN F, SUN L, KARDIAN D B, et al . Feedback inhibition of calcineurin and Ras by a dual inhibitory protein Carabin[J ] . Nature, 2007, 445(7126): 433-436. DOI: 10.1038/nature05476.

BISSERIER M, BERTHOUZE-DUQUESNES M, BRECKLER M, et al . Carabin protects against cardiac hypertrophy by blocking calcineurin, ras, and Ca 2+ /calmodulin-dependent protein kinase Ⅱ signaling[J ] . Circulation, 2015, 131(4): 390-400. DOI: 10.1161/CIRCULATIONAHA.114.010686..

ZHU X, FANG J, GONG J, et al . Cardiac-specific EPI64C blunts pressure overload-induced cardiac hypertrophy[J ] . Hypertension, 2016, 67(5): 866-877. DOI: 10.1161/HYPERTENSIONAHA.115.07042.

VOLLAND C, BREMER S, HELLENKAMP K, et al . Enhanced cardi ac TBC1D10C expression lowers heart rate and enhances exercise capacity and survival[J ] . Sci Rep, 2016, 6: 33853. DOI: 10.1038/srep33853.

DAVIDSON M M, NESTI C, PALENZUELA L, et al . Novel cell lines derived from adult human ventricular cardiomyocytes[J ] . J Mol Cell Cardiol, 2005, 39(1): 133-147. DOI: 10.1016/j.yjmcc.2005.03.003.

刘凯玉, 蒋维, 王瑞娜, 等.人Carabin蛋白过表达慢病毒载体构建及其稳定表达细胞株的筛选[J].生物学杂志, 2016, 33(3): 1-4. DOI: 10.3969/j.issn.2095-1736.2016.03.001.

LIU K Y, JIAN W, WANG R N, et al . Construction of lentivirus expression vector of human CARABIN and screening of cell line stably expressing CARABIN[J ] . J Biol, 2016, 33(3): 1-4. DOI: 10.3969/j.issn.2095-1736.2016.03.001.

ROTH G A, JOHNSON C, ABAJOBIR A, et al . Global, regional, and national burden of cardiovascular diseases for 10 causes, 1990 to 2015[J ] . J Am Coll Cardiol, 2017, 70(1): 1-25. DOI: 10.1016/j.jacc.2017.04.052.

DIAS L D, CASALI K R, GHEM C, et al . Mesenchymal stem cells from sternum: the type of heart disease, ischemic or valvular, does not influence the cell culture establishment and growth kinetics[J ] . J Transl Med, 2017, 15(1): 161-170. DOI: 10.1186/s12967-017-1262-0.

NI H, XU J. Recent trends in heart failure-related mortality: United States, 2000-2014[J]. NCHS Data Brief, 2015 (231): 1-8.

LALIT P A, HEI D J, RAVAL A N, et al . Induced pluripotent stem cells for post-myocardial infarction repair: remarkable opportunities and challenges[J ] . Circ Res, 2014, 114(8): 1328-1345. DOI: 10.1161/CIRCRESAHA.114.300556.

MOON S H, KANG S W, PARK S J, et al . The use of aggregates of purified cardiomyocytesderived from human ESCs for functionalengraftment after myocardial infarction[J ] . Biomaterials, 2013, 34(16): 4013-4026. DOI: 10.1016/j.biomaterials.2013.02.022.

AILAWADI S, WANG X, GU H, et al . Pathologic function and therapeutic potential of exosomes in cardiovascular disease[J ] . Biochim Biophys Acta, 2015, 1852(1): 1-11. DOI: 10.1016/j.bbadis.2014.10.008.

SCHICKEL J N, PASQUALI J L, SOLEY A, et al . Carabin deficiency in B cells increases BCR-TLR9 costimulation-induced autoimmunity[J ] . EMBO Mol Med, 2012, 4(12): 1261-1275. DOI: 10.1002/emmm.201201595.

HSU C, MOROHASHI Y, YOSHIMURA S, et al . Regulation of exosome secretion by Rab35 and its GTPase-activating proteins TBC1D10C[J ] . J Cell Biol, 2010, 189(2): 223-232. DOI: 10.1083/jcb.200911018.

JIANG W, ZHOU X, LI Z, et al . Prolyl 4-hydroxylase 2 promotes B-cell lymphoma progression via hydroxylation of Carabin[J ] . Blood, 2018, 131(12): 1325-1336. DOI: 10.1182/blood-2017-07-794875..

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