miR-186-5p is Expressed Highly in Ethanol-induced Cardiomyocytes and Regulates Apoptosis by Target Gene <i>XIAP</i>

doi:10.13523/j.cb.20190506

China Biotechnology

2019, Vol. 39

Issue (5): 53-62 DOI: 10.13523/j.cb.20190506

miR-186-5p is Expressed Highly in Ethanol-induced Cardiomyocytes and Regulates Apoptosis by Target Gene XIAP

Ye LIU(

),Yue PAN,Wei ZHENG,Jing HU

Electrocardial Center of the First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000, China

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Abstract

Objective: To confirm the effect of alcohol-induced apoptosis of AC16 cardiomyocytes and the relationship with the alcohol concentration and the action time.To investigate the expression levels of miR-186-5p and X-linked inhibitor of apoptosis protein(XIAP) along with the variational apoptosis level in alcohol -treated AC16 cardiomyocytes with different concentrations. To explore miR-186-5p regulated apoptosis of alcohol-induced cardiomyocytes using XIAP as the target gene.Methods:Flow cytometry analyzed the expression levels of cardiomyocyte apoptosis. Western blot and real-time PCR detected respectly the protein and RNA expression levels of miR-186-5p and XIAP in alcohol-treated AC16 cardiomyocytes. Luciferase reporter gene experiment verified that XIAP is a direct target gene of miR-186-5p.Results:Alcohol induced cardiomyocyte apoptosis, and the levels of apoptosis were dependent on alcohol concentration and duration of time. Alcohol intake can increase the expression levels of miR-186-5p and decrease the expression levels of XIAP in AC16 cardiomyocytes. miR-186-5p was involved in alcohol-induced the process of AC16 cardiomyocyte apoptosis.XIAP can inhibite alcohol-induced AC16 cardiomyocyte apoptosis.miR-186-5p targets XIAP as a target gene to regulate alcohol-induced cardiomyocyte apoptosis.Conclusion:The levels of apoptosis in AC16 cardiomyocytes increase after alcohol treatment and increase further with the increase of alcohol concentration and the prolongation of the action time. The expression levels of miR-186-5p increase, and the expression levels of XIAP decrease in alcohol-treated AC16 cardiomyocytes. miR-186-5p regulates apoptosis of alcohol-induced cardiomyocytes using XIAP as the target gene.

Key words： Cardiomyocyte Apoptosis miR-186-5p XIAP Alcoholic cardiomyopathy

Received: 24 October 2018 Published: 04 June 2019

ZTFLH:

R542.2

Corresponding Authors: Ye LIU E-mail: 601924547@qq.com

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	Ye LIU
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Cite this article:

Ye LIU,Yue PAN,Wei ZHENG,Jing HU. miR-186-5p is Expressed Highly in Ethanol-induced Cardiomyocytes and Regulates Apoptosis by Target Gene XIAP. China Biotechnology, 2019, 39(5): 53-62.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20190506 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I5/53

Fig.1 Flow cytometry analyzes the expression levels of alcohol-induced cardiomyocyte apoptosis Q4-1: The cells in this area are necrotic cells and mechanical injury-cells;Q4-2: The cells in this area are late-stage apoptotic cells; Q4-3: Cells in this area are living cells; Q4-4: Cells in this area are early-stage apoptotic cells

Fig.2 Luciferase reporter gene experiment verifies that XIAP is a direct target gene of miR-186-5p The results showed fluorescence intensity in the group of the wild-type pmirGLO-wt-XIAP-3'-UTR combined with miR-186-5p mimic decreased compared with the other three groups, which showed a statistically significant difference (* P<0.05 vs the other three groups), and no significant difference was observed in fluorescence intensity among the other three groups(P>0.05)

Fig.3 Real-time PCR detects the expression levels of miR-186-5p in alcohol-treated AC16 cardiomyocytes

Fig.4 Western blot and real-time PCR detect the expression levels of XIAP in alcohol-treated AC16 cardiomyocytes

Fig.5 Flow cytometry analyzes transfecting miR-186-5p mimic affects the levels of alcohol-induced apoptosis of cardiomyocytes Q2-1: The cells in this area are necrotic cells and mechanical injury-cells;Q2-2: The cells in this area are late -stage apoptotic cells;Q2-3: Cells in this area are living cells;Q2-4: Cells in this area are early-stage apoptotic cells

Fig.6 Flow cytometry analyzes transfecting XIAP plasmid affects the levels of alcohol-induced apoptosis of cardiomyocytes Q2-1: The cells in this area are necrotic cells and mechanical injury-cells;Q2-2: The cells in this area are late-stage apoptotic cells;Q2-3: Cells in this area are living cells;Q2-4: Cells in this area are early-stage apoptotic cells

Fig.7 Western blot and real-time PCR detect the expression levels of XIAP in cardiomyocytes transfected with miR-186-5p mimic/inhibitor


[1]	Naimi T S, Nelson D E, Brewer R D , et al. The intensity of binge alcohol consumption among U.S.adults. American Journal of Preventive Medicine, 2010,38(2):201-207. doi: 10.1016/j.amepre.2009.09.039

[2]	Steiner J L, Lang C H . Etiology of alcoholic cardiomyopathy:Mitochondria,oxidative stress and apoptosis. Int J Biochem Cell Biol, 2017,8(89):125-135.

[3]	Wang Z, Song J, Zhang L , et al. Increased expression of microRNA-378a-5p in acute ethanol exposure of rat cardiomyocytes. Cell Stress Chaperones, 2017,22(2):245-252. doi: 10.1007/s12192-016-0760-y

[4]	Jing L, Lin C, Lu Y , et al. Investigation of microRNA expression profiles associated with human Alcoholic cardiomyopathy. Cardiology, 2015,130(4):223-233. doi: 10.1159/000370028

[5]	Lai Y, Guo H, Li J , et al. Comparison of surgical results in patients with hypertrophic obstructive cardiomyopathy after classic or modified morrow septal yectomy. Medicine (Baltimore), 2017,96(51):9371. doi: 10.1097/MD.0000000000009371

[6]	Jiang J, Mo H, Liu C , et al. Inhibition of miR-186-5p contributes to high glucose-induced injury in AC16 cardiomyocytes. Exp Ther Med, 2018,15(1):627-632.

[7]	Bradford M M . A rapid and sensitive method for the quantitationof microgram quantities of protein utilizing the principle of protein-dye binding.Anal Biochem, 1976(72):248-254.

[8]	Ji F, Liu Q, Feng Z . Genetic association between 1425G/A SNP in PRKCH and hypertrophic cardiomyopathy in a Chinese population. Oncotarget, 2017,8(70):114839-114844.

[9]	Mu J, Zhang G, Xue D , et al. Sudden cardiac death owing to arrhythmogenic right ventri-cularcardiamyopathy.Two case reports and systematic literature review. Medicine(Baltimore), 2017,96(47):8808.

[10]	Dahraoui S, Uwingabiye J, Belarj B , et al. Unexpected discovery of multiple myeloma following cardiomyopathy. Clin Case Rep, 2018,6(1):86-90. doi: 10.1002/ccr3.2018.6.issue-1

[11]	Adachi K, ashiguchi S, Saito M , et al. Detection and management of cardiomyopathy in female dystrophinopathy carriers. J Neurol Sci, 2018,3(386):74-80.

[12]	Baltrūniené V, Bironaité D, Kažukauskiené I , et al. The role of serum adiponectin for outcome prediction in patients with dilated cardiomyopathy and advanced heart failure. [2019-04-09]. https://www.hindawi.com/journals/bmri/2017/3818292/

[13]	Bollen I, Van der Meulen M, De G K, ,et al.Cardiomyocyte hypocontractility and reduced myofibril density in end stage pediatric cardiomyopathy. Front Physiol, 2017,12(8):1-12.

[14]	Buccheri D, Zambelli G . The link between spontaneous coronary artery dissection and takosubo cardiomyopathy.analysis of the published cases. J Thorac Dis, 2017,9(12):5489-5492. doi: 10.21037/jtd

[15]	Fu K Y, Zamudio R, Henderson-Frost J , et al. Association of capase-1 polymorphisms with Chagas cardiomyopathy among individuals in Santa Cruz,Bolivia. Rev Soc Bras Med Trop, 2017,50(4); 516-523. doi: 10.1590/0037-8682-0015-2017

[16]	Edison N, Curtz Y, Paland N , et al. Degradation of Bcl-2 by XIAP and ARTS Promotes Apoptosis. Cell Rep, 2017,21(2):442-454. doi: 10.1016/j.celrep.2017.09.052

[17]	Cesa L C, Shao H, Srinivasan S R , et al. X-Linked inhibitor of apoptosis protein(XIAP) is a client of heat shock protein 70(Hsp 70) and a biomarker of its inhibition. J Biol Chem, 2018,16(2):7.

[18]	Yang W, Zhou H, Yan Y . XIAP underlies apoptosis resistance of renal cell carcinoma cells. Mol Med Rep, 2018,17(1):125-130.

[19]	Edison N, Curtz Y, Paland N , et al. Degradation of Bcl-2 by XIAP and ARTS promotes apoptosis. Cell Rep, 2017,21(2):442-454. doi: 10.1016/j.celrep.2017.09.052

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