1. 佳木斯大学研究生学院, 黑龙江 佳木斯 154007; 2. 齐齐哈尔医学院, a. 公共卫生学院, b. 医药科学研究院, 黑龙江 齐齐哈尔 161006
Effect of Trans-oleic Acid on Proliferation Inhibition and Apoptosis Induction in H9c2 Cardiomyocyte
CHEN Zhe1,2a, ZHU Shuai-wei2a, MA Li-wei2b, LI Hong-long2a, XUE Hai-feng2a, LI Ji-yuan2a, ZHAO Peng2a*, LOU Feng-ge2a*
1. Graduate School, Jiamusi University, Jiamusi 154007, China; 2a. School of Public Health, 2b. Research Institute of Medicine & Pharmacy, Qiqihar Medical University, Qiqihar 161006, China
Abstract:OBJECTIVE To investigate the effects and possible mechanism of trans-oleic acid (9t-C18:1) on proliferation inhibition and induction apoptosis in H9c2 cardiomyocyte. METHODS H9c2 rat cardiomyocytes were cultured in vitro. High, medium and low (600, 300, 150 μmol·L-1) dose of 9t-C18:1 groups and the negative control (NC) group were administered to H9c2 cardiomyocytes. The effect of 9t-C18:1 on cell proliferation was tested using cell counting kit-8 (CCK-8) assay. Morphological changes of cells were observed by AO-EB staining. Intracellular reactive oxygen species (ROS) and apoptosis were detected by flow cytometry. The expression of Bcl-2 and Bax genes was detected by quantitative real time- polymerase chain reaction (QRT-PCR). The expression of Bcl-2 and Bax protein was determined by flow cytometry after immunofluorescence staining. RESULTS The typical morphological characteristics of apoptosis were observed by fluorescence microscope. The result of CCK-8 assay indicated that 9t-C18:1 have an certainly inhibitory effect on the proliferation of H9c2 cells. ROS level and apoptosis rate were significantly increased. Bcl-2 gene and protein expression were down-regulated, and Bax gene and protein expression were up-regulated, compared with NC group(P<0.05, P<0.01). CONCLUSION 9t-C18:1 can inhibit the proliferation and induce the apoptosis of H9c2 cardiomyocyte, and its mechanism may be related to promoting the mitochondrial apoptotic pathway.
SHEN J F, ZHANG Z Y. Safety of trans fatty acids and recent research progress [J]. J Chin Cere Oils Assoc(中国粮油学报), 2005, 20(4):88-90.
[2]
COUNIL E, JULIEN P, LAMARCHEB, et al. Association between trans-fatty acids in erythrocytes and pro-atherogenic lipid profiles among Canadian Inuit of Nunavik possible influences of sex and age [J]. Br J Nutr, 2009, 102(5):766-776.
[3]
LI X P. Effect of trans fatty acid can proliferation and function of human umbilical vein smooth muscle cells [D]. Nanchang: Nanchang University,2013.
[4]
ROZENN N L, IRENA B K, TRIVELLORE E R. Cell membrane trans fatty acids and the risk of primary cardic arrest [J]. Circulation, 2002, 105:697-701.
[5]
MOZAFFARIAN D, ARO A, WILLETT W C. Health effects of trans-fatty acids: experimental and observational evidence [J]. Eur J Clin Nutr, 2009, 63(suppl 2):5-21.
[6]
OOMEN C M, OCKE M C, FESKENS E J, et al. Association between trans fatty acids intake and 10-year risk of coronary heart disease in the Zutphen Elderly Study a prospective population-based study [J]. Lancet, 2001, 357(9258):746-751.
[7]
DOWNAR D Z, KOSMIDER A, NARUSZEWICZ M. Trans fatty acids induce apoptosis in human endothelial cells [J]. Physiol Pharmacol, 2005, 56(4):611-625.
[8]
KONDOH Y, KAWADA T, URADE R. Activation of caspase-3 in HepG2 cells by elaidic acid (t18:1) [J]. Biochim Biophys Acta, 2007, 1771(4):500-505.
[9]
BAINES C P, MOLKENTIN J D. Stress signaling pathways that modulate cardiac myocyte apoptosis [J]. J Mol Cell Cardiol, 2005, 38(1):47-62.
[10]
WANG L W, LIU F Z, LU J K, et al. Effect of flavonols on caspase-3, Bcl-2 and Bax expression in cardiomyocytes apoptosis [J]. Chin Pharm J(中国药学杂志), 2007, 42(7):501-504.
[11]
KIMES B W, BRANDT B L. Properties of clonal muscle cell line from rat heart [J]. Exp Cell Res, 1976, 98(2):367-381.
[12]
JOSE CORBALAN J, VATNER D E, VATNE R S F. Myocardial apoptosis in heart disease: does the emperor have clothes? [J]. Basic Res Cardiol, 2016, 111:31.
[13]
KUZMICIC J, DEL CAMPO A, LPEZ-CRISOSTO C, et al. Mitochondrial dynamics: a potential new therapeutic target for heart failure [J]. Rev Esp Cardiol, 2011, 64 (10):916-923.
[14]
BOTT-FLGEL L, WEIG H J, UHLEIN H, et al. Quantitative analysis of apoptotic markers in human end-stage heart failure [J]. Eur J Heart Fail, 2008, 10(2):129-132.
[15]
TSIPIS A, ATHANASSIADOU A M, ATHANASSIADOU P, et al. Apoptosis-related factors p53, bcl-2 and the defects of force transmission in dilated cardiomyopathy [J]. Pathol Res Pract, 2010, 206(9):625-630.
[16]
FENG W S, YANG F F, ZHANG L, et al. Protective effects of aqueous extract from descurainia sophia on doxorubicin-induced cardiomyocyte injuryvia suppressing apoptosis and oxidative stress [J]. Chin Pharm J(中国药学杂志), 2018, 53(23):1999-2007.
[17]
BI L, YAN X, CHEN W, et al. Antihepatocellular carcinoma potential of tetramethylpyrazine induces cell cycle modulation and mitochondrial-dependent apoptosis: regulation of p53 signaling pathway in Hep G2 cells in vitro [J]. Integr Cancer Ther, 2016, 15(2):226-236.
[18]
MA L W, ZHOU L, LI S L, et al. Effect of 12-deoxyphorbol-13-palmitate on proliferation inhibition and apoptosis induction of HL60 cells [J]. Chin Pharm J(中国药学杂志),2018,53 (1):30-34.
[19]
SONG R S, XIAO X H, LIU Z L, et al. Effects of a monomer purified from paris polyphylla (PP-22) on proliferation and apoptosis of human gastric carcinoma MGC803 cells [J]. Chin Pharm J(中国药学杂志), 2015, 50(18):1600-1606.
[20]
LIU H C, ZHANG Y, ZHANG S, et al. Correlation research on the protein expression (p75 NTR, bax, bcl-2 and caspase-3) and cortical neuron apoptosis following mechanical injury in rat [J]. Eur Rev Med Pharmacol Sci, 2015, 19(18):3459-3467.
[21]
WANG S J, GUO X, ZUO H, et al. Chondrocyte apoptosis and expression of Bcl-2, Bax, Fas, and iNOS in articular cartilage in patients with Kashin-Beck disease [J]. J Rheumatol, 2006, 33(3):615-619.