Sains Malaysiana 53(5)(2024): 1185-1199

http://doi.org/10.17576/jsm-2024-5305-16

Lipoprotein Oxidation, Genetic Expression and Polymorphism of Apo A and B Gene in Patients with Coronary Artery Disease in a Pakistani Punjabi Population

(Pengoksidaan Lipoprotein, Ekspresi Genetik dan Polimorfisme Gen Apo A dan B pada Pesakit Penyakit Arteri Koronari dalam Kalangan Penduduk Pakistani Punjabi)

HIRA SOHAIL^1,2,*, MADEEHA SHAHZAD², SABA KHALIQ³ & FARHAT IJAZ¹

¹Combined Military Hospital Lahore Medical College and Institute of Dentistry, Lahore, Pakistan

²The University of Lahore, Pakistan

³University of Health Sciences, Lahore, Pakistan

Diserahkan: 27 September 2023/Diterima: 20 Mac 2024

Abstract

Oxidative alterations of lipoproteins and numerous polymorphism locations in the apolipoprotein A1 (APOA1) and B (APOB) genes have been identified as risk factors for cardiovascular events. The aim of the study was to investigate the gene expression and the role of APOA and B gene polymorphisms in coronary artery disease (CAD) and their relationship with lipid profiles, oxidized High Density Lipoprotein (oxHDL), and oxidized Low Density Lipoprotein (oxLDL) in the Punjabi Pakistani population. A total of 200 subjects were included, of which 135 were diagnosed as having CAD and 65 were not. The lipid profile, oxHDL, and oxLDL were measured through the calorimetric method and ELISA, respectively. Quantitative variables represented by mean ± SD for normally distributed data. Gene expression was done on cDNA through Real-time PCR. Genotyping of APOA1 (rs670, rs5069) and APOB (rs693, rs11279109) were performed by PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism), and gel electrophoresis was used for band visualization. Our findings showed that the oxHDL, oxLDL, APOA, B gene expression and Del genotype of APOB rs11279109 was significantly increased in CAD cases. In addition, significant high levels of oxHDL, oxLDL, serum cholesterol, VLDL, TG, and cholesterol/HDL ratio has been shown to be associated with CAD. APOB (rs11279109) has a significant impact on dyslipidemia and exhibit a higher risk of CAD (OR=3.97) while, APOB (rs693), APOA1 (rs670), (rs5069) possesses a lower risk of CAD in the population. Hence, levels of oxHDL and oxLDL as well as ApoA1 and APOB genotyping could be useful in identifying individuals who are at risk of CAD.

Keywords: APOA1 gene; APOB gene; coronary artery disease; dyslipidemia; oxHDL; oxLDL

Abstrak

Perubahan oksidatif lipoprotein dan banyak lokasi polimorfisme dalam gen apolipoprotein A1 (APOA1) dan B (APOB) telah dikenal pasti sebagai faktor risiko gangguan kardiovaskular. Matlamat penyelidikan ini adalah untuk mengkaji pengekspresan gen dan peranan polimorfisme gen APOA dan B dalam penyakit arteri koronari (CAD) dan hubungannya dengan profil lipid, lipoprotein ketumpatan tinggi (oxHDL) teroksida dan lipoprotein ketumpatan rendah (oxLDL) teroksida dalam kalangan penduduk Punjabi Pakistan. Sebanyak 200 subjek telah dimasukkan dalam kajian ini dengan 135 daripadanya telah didiagnosis sebagai mempunyai CAD dan 65 tidak. Profil lipid, oxHDL dan oxLDL diukur masing-masing melalui kaedah kalorimetrik dan ELISA. Pemboleh ubah kuantitatif diwakili oleh min ± SD untuk data taburan normal. Pengekspresan gen dilakukan pada cDNA melalui PCR masa nyata. Genotaip APOA1 (rs670, rs5069) dan APOB (rs693, rs11279109) telah dilakukan oleh PCR-RFLP (Tindak Balas Berantai Polimerase - Polimorfisme Pemotongan Panjang Cebisan (PCR-RFLP)) dan elektroforesis gel digunakan untuk visualisasi jalur. Penemuan kami menunjukkan bahawa pengekspresan gen oxHDL, oxLDL, APOA, B dan genotip Del APOB rs11279109 meningkat dengan ketara dalam kes CAD. Di samping itu, paras tinggi oxHDL, oxLDL, kolesterol serum, VLDL, TG dan nisbah kolesterol/HDL yang tinggi telah terbukti dikaitkan dengan CAD. APOB (rs11279109) mempunyai kesan yang ketara terhadap dislipidemia dan menunjukkan risiko CAD yang lebih tinggi (OR=3.97) manakala, APOB (rs693), APOA1 (rs670), (rs5069) mempunyai risiko CAD yang lebih rendah dalam populasi. Oleh itu, tahap oxHDL dan oxLDL serta genotaip ApoA1 dan APOB boleh berguna dalam mengenal pasti individu yang berisiko mendapat CAD.

Kata kunci: Dislipidemia; gen APOA1; gen APOB; oxHDL; oxLDL; penyakit arteri koronari

RUJUKAN

Abd El-Aziz, T.A. & Mohamed, R.H. 2016. LDLR, ApoB and ApoE genes polymorphisms and classical risk factors in premature coronary artery disease. Gene 590(2): 263-269. https://doi.org/10.1016/j.gene.2016.05.032

Al-Bustan, S.A., Alnaqeeb, M.A., Annice, B.G., Ebrahim, G.A. & Refai, T.M. 2014. Genetic association of APOB polymorphisms with variation in serum lipid profile among the Kuwait population. Lipids in Health and Disease 13: 157. DOI: 10.1186/1476-511X-13-157

Al-Bustan, S.A., Al-Serri, A.E., Annice, B.G., Alnaqeeb, M.A. & Ebrahim, G.A. 2013. Re-sequencing of the APOAI promoter region and the genetic association of the-75G> A polymorphism with increased cholesterol and low density lipoprotein levels among a sample of the Kuwaiti population. BMC Medical Genetics 14: 90. doi:10.1186/1471-2350-14-90

Boekholdt, S.M., Peters, R.J., Fountoulaki, K., Kastelein, J.J. & Sijbrands, E.J. 2003. Molecular variation at the apolipoprotein B gene locus in relation to lipids and cardiovascular disease: A systematic meta-analysis. Human Genetics 113: 417-425. DOI: 10.1007/s00439-003-0988-3

Bogari, N.M., Abdel-Latif, A.M., Hassan, M.A., Ramadan, A. & Fawzy, A. 2015. No association of apolipoprotein B gene polymorphism and blood lipids in obese Egyptian subjects. Journal of Negative Results in Biomedicine 14: 7. doi: 10.1186/s12952- 015-0026-8

Burgos Alves, M.I., Avilés Plaza, F., Martínez-Tomás, R., Sánchez-Campillo, M., Larqué, E., Pérez-Llamas, F., Martínez Hernández, P. & Parra Pallarés, S. 2010. Oxidized LDL and its correlation with lipid profile and oxidative stress biomarkers in young healthy Spanish subjects. Journal of Physiology and Biochemistry 66: 221-227. DOI: 10.1007/s13105-010-0028-4

Casillas-Munoz, F., Valle, Y., Muñoz-Valle, J.F., Martínez-Fernández, D.E., Reynoso-Villalpando, G.L., Flores-Salinas, H.E., Llamas-Covarrubias, M.A. & Padilla-Gutiérrez, J.R. 2018. APOA1 and APOB polymorphisms and apolipoprotein concentrations as biomarkers of risk in acute coronary syndrome: Relationship with lipid-lowering therapy effectiveness. Medicina Clínica (English Edition) 151(1): 1-7. doi:10.1016/j.medcle.2018.05.001

Cavalli, S.A., Hirata, M.H., Salazar, L.A., Diament, J., Forti, N., Giannini, S.D., Nakandakare, E.R., Bertolami, M.C. & Hirata, R.D. 2000. Apolipoprotein B gene polymorphisms: Prevalence and impact on serum lipid concentrations in hypercholesterolemic individuals from Brazil. Clinica Chimica Acta 302(1-2): 189-203. https://doi.org/10.1016/S0009-8981(00)00367-3

Chen, Y., Lin, M., Liang, Y., Zhang, N. & Rao, S. 2016. Association between apolipoprotein b XbaI polymorphism and coronary heart disease in Han Chinese population: A meta-analysis. Genetic Testing and Molecular Biomarkers 20(6): 304-311. DOI: 10.1089/gtmb.2015.0126

Chen, E.S., Mazzotti, D.R., Furuya, T.K., Cendoroglo, M.S., Ramos, L.R., Araujo, L.Q., Burbano, R.R. & de Arruda Cardoso Smith, M. 2009. Apolipoprotein A1 gene polymorphisms as risk factors for hypertension and obesity. Clinical and Experimental Medicine 9: 319-325. https://doi.org/10.1007/s10238-009-0051-3

Chen, S., Xie, X., Wang, Y., Gao, Y., Xie, X., Yang, J. & Ye, J. 2014. Association between leukocyte mitochondrial DNA content and risk of coronary heart disease: A case-control study. Atherosclerosis 237(1): 220-226. doi:10.1016/j.atherosclerosis.2014.08.051

Chiodini, B.D., Barlera, S., Franzosi, M.G., Beceiro, V.L., Introna, M. & Tognoni, G. 2003. APO B gene polymorphisms and coronary artery disease: A meta-analysis. Atherosclerosis 167(2): 355-366. DOI: 10.1016/S0021-9150(02)00425-2

Dawar, R., Gurtoo, A. & Singh, R. 2010. Apolipoprotein A1 gene polymorphism (G–75A and C+ 83T) in patients with myocardial infarction: A pilot study in a north Indian population. American Journal of Clinical Pathology 134(2):  249-255. doi:10.1309/AJCPKPTXQ3QN1IFG

Dong, L.P. & Da Huang, W.J.C. 2018. Meta-analysis of the association between Apo-A1 rs670, rs5069 polymorphisms and coronary artery diseases. International Journal of Clinical and Experimental Medicine 11(7): 6445-6453.

Duman, B.S., Türkoğlu, Ç., Akpinar, B., Güden, M., Vertii, A., Çağatay, P., Günay, D. & Sevim Büyükdevrim, A. 2005. Genetic variations of the apolipoprotein B gene in Turkish patients with coronary artery disease. Annals of Human Biology 32(5): 620-629. https://doi.org/10.1080/03014460500228626

El-Sayed, K., Almenshawy, A., El-Farahaty, R.M., Osman, A.M. & Mahsoub, N. 2022. Association of XbaI polymorphism of ApoB gene with lipid profile, risk factors and dietary pattern in coronary artery disease Egyptian rural and urban patients. International Journal of Health Sciences (Qassim) 6: 11251. DOI: 10.53730/ijhs.v6nS5.11251

Friedewald, W.T., Levy, R.I. & Fredrickson, D.S. 1972. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clinical Chemistry 18(6): 499-502. https://doi.org/10.1093/clinchem/18.6.499

Gu, W., Zhang, M. & Wen, S. 2015. Association between the APOB XbaI and EcoRI polymorphisms and lipids in Chinese: A meta-analysis. Lipids in Health and Disease 14: 123. DOI: 10.1186/s12944-015-0125-z

Han, Y., Yang, C.K., Gao, C.Y., Liu, Y.H., Cheng, J.T., Wang, X.P., Wang, Z.M. & Cheng, T.F. 2017. Association between the ApoB/A1 ratio and the vulnerability of LMCA plaque in the patients with stable angina pectoris. Zhonghua Yi Xue Za Zhi 97(27): 2101-2106. DOI: 10.3760/cma.j.issn.0376-2491.2017.27.005

Hanif, M.K., Fan, Y., Wang, L., Jiang, H., Li, Z., Ma, M., Ma, L. and Ma, M. 2022. Dietary habits of patients with coronary artery disease: A case-control study from Pakistan. International Journal of Environmental Research and Public Health 19(14): 8635. https://doi.org/10.3390/ijerph19148635

Hassan, N.E., El-Masry, S.A., Zarouk, W.A., Abd Elneam, A.I., Rasheed, E.A. & Mahmoud, M.M. 2015. Apolipoprotein B polymorphism distribution among a sample of obese Egyptian females with visceral obesity and its influence on lipid profile. Journal of Genetic Engineering and Biotechnology 13(2): 177-183. doi: 10.1016/ j.jgeb.2015.09.005

Honda, H., Ueda, M., Kojima, S., Mashiba, S., Suzuki, H., Hosaka, N., Hirai, Y., Nakamura, M., Nagai, H., Kato, N., Mukai, M., Watanabe, M., Takahashi, K., Shishido, K. & Akizawa, T. 2010. Oxidized high-density lipoprotein is associated with protein-energy wasting in maintenance hemodialysis patients. Clinical Journal of the American Society of Nephrology 5(6): 1021-1028.

Hosseini-Esfahani, F., Mirmiran, P., Daneshpour, M.S., Mottaghi, A. & Azizi, F. 2017. The effect of interactions of single nucleotide polymorphisms of APOA1/APOC3 with food group intakes on the risk of metabolic syndrome. Avicenna Journal of Medical Biotechnology 9(2): 94-103.

Itabe, H. & Obama, T. 2023. The oxidized lipoproteins in vivo: Its diversity and behavior in the human circulation. International Journal of Molecular Sciences 24(6): 5747. https://doi.org/10.3390/ijms24065747

Lamia, R., Asma, O., Slim, K., Jihene, R., Imen, B., Ibtihel, B.H., Kaouther, K., Radhia, B., Nabila, B.R., Naoufel, N. & Ahmed, B.A. 2012. Association of four apolipoprotein B polymorphisms with lipid profile and stenosis in Tunisian coronary patients. Journal of Genetics 91(1): 75-79.

Liao, B., Cheng, K., Dong, S., Liu, H. & Xu, Z. 2015. Effect of apolipoprotein A1 genetic polymorphisms on lipid profiles and the risk of coronary artery disease. Diagnostic Pathology 10: 102. DOI: 10.1186/s13000-015-0328-7

Liting, P., Guoping, L. & Zhenyue, C. 2015. Apolipoprotein B/apolipoprotein A1 ratio and non-high-density lipoprotein cholesterol. Herz 40(Suppl 1): 1-7. https://doi.org/10.1007/s00059-014-4147-5

Liu, Y.L., Zhang, Y.B., Li, Y., Ma, R.L., Cai, W.W., Lin-Jiang, L., Wang, T.S. & Yao, Z. 2014. Correlation between the Xba I polymorphism of apoB gene and serum lipid profiles in Li ethnic group. Asian Pacific Journal of Tropical Medicine 7(1): 63-66. doi:10.1016/S1995-7645(13)60193-5

Ma, J., Zhang, H.Q. & Wang, W. 2011. Relationship between apolipoprotein B, apolipoprotein E and angiotensinogen gene polymorphisms and coronary heart disease. China Modern Doctor 49: 4-6.

Nikolajevic Starcevic, J., Santl Letonja, M., Praznikar, Z.J., Makuc, J., Vujkovac, A.C. & Petrovic, D. 2014. Polymorphisms XbaI (rs693) and EcoRI (rs1042031) of the ApoB gene are associated with carotid plaques but not with carotid intimamedia thickness in patients with diabetes mellitus type 2. Vasa 43: 171-180. doi:10.1024/0301-1526/a000346

Niu, C., Luo, Z., Yu, L., Yang, Y., Chen, Y., Luo, X., Lai, F. & Song, Y. 2017. Associations of the APOB rs693 and rs17240441 polymorphisms with plasma APOB and lipid levels: A meta-analysis. Lipids In Health and Disease 16: 1-20. doi:10.1186/s12944-017-0558-7

Pagonas, N., Mueller, R., Weiland, L., Jaensch, M., Westhoff, T.H., Buschmann, I.R., Sasko, B., Ritter, O. & Kelesidis, T.H. 2020. Oxidation of HDL in patients with coronary artery disease. European Heart Journal 41(Supplement_2): ehaa946-1521. DOI: https://doi.org/10.1093/ehjci/ehaa946.1521

Pan, J.P., Chiang, A.N., Tai, J.J., Wang, S.P. & Chang, M.S. 1995. Restriction fragment length polymorphisms of apolipoprotein B gene in Chinese population with coronary heart disease. Clinical Chemistry 41: 424-429. https://doi.org/10.1093/clinchem/41.3.424

Rader, D.J. & Hovingh, G.K. 2014. HDL and cardiovascular disease. Lancet 384: 618-625. DOI: 10.1016/S0140-6736(14)61217-4

Rafiee, M., Sotoudeh, G., Djalali, M., Alvandi, E., Eshraghian, M., Javadi, F., Doostan, F. & Koohdani, F. 2019. The interaction between apolipoprotein B insertion/deletion polymorphism and macronutrient intake on lipid profile and serum leptin and ghrelin levels in type 2 diabetes mellitus patients. European Journal of Nutrition 58: 1055-1065. DOI: 10.1007/s00394-018-1621-5

Rebhi, L., Omezzine, A., Kchok, K., Belkahla, R., Ben HadjMbarek, I., Rejeb, J., Ben Rejeb, N., Nabli, N., Bibi, A., Massoud, T. & Abdelaziz, A. 2008. 5′ ins/del and 3′ VNTR polymorphisms in the apolipoprotein B gene in relation to lipids and coronary artery disease. Clinical Chemical Laboratory Medicine 46: 329-334. https://doi.org/10.1515/CCLM.2008.067

Roberts, R., Chang, C.C. & Hadley, T. 2021. Genetic risk stratification: A paradigm shift in prevention of coronary artery disease. Journal of American College Cardiology Basic to Translational Science 6(3): 287-304. https://doi.org/10.1016/j.jacbts.2020.09.004

Rodrigues, A.C., Sobrino, B., Genvigir, F.D.V., Willrich, M.A.V., Arazi, S.S., Dorea, E.L., Bernik, M.M.S., Bertolami, M., Faludi, A.A., Brion, M.J. & Carracedo, A. 2013. Genetic variants in genes related to lipid metabolism and atherosclerosis, dyslipidemia and atorvastatin response. Clinica Chimica Acta 417: 8-11. doi:10.1016/j.cca.2012.11.028

Segrest, J.P., Li, L., Anantharamaiah, G.M., Harvey, S.C., Liadaki, K.N. & Zannis, V. 2000. Structure and function of apolipoprotein A-I and high-density lipoprotein. Current Opinion in Lipidology 11: 105-115.

Shanker, J., Perumal, G., Rao, V.S., Khadrinarasimhiah, N.B., John, S., Hebbagodi, S., Mukherjee, M. & Kakkar, V.V. 2008. Genetic studies on the APOA1-C3-A5 gene cluster in Asian Indians with premature coronary artery disease. Lipids In Health and Disease 7: 33. DOI: 10.1186/1476-511X-7-33

Sharma, R., Mahajan, M., Singh, B., Singh, G. & Singh, P. 2011. Role of the APOB gene polymorphism (c.12669G>A, p. Gln4154Lys) in coronary artery disease in the Indian Punjabi Population. Balkan Journal of Medical Genetics 14: 35-40. doi:10.2478/v10034-011-0045-9

Sorokin, A.V., Kotani, K., Elnabawi, Y.A., Dey, A.K., Sajja, A.P., Yamada, S., Ueda, M., Harrington, C.L., Baumer, Y., Rodante, J.A. & Gelfand, J.M. 2018. Association between oxidation-modified lipoproteins and coronary plaque in psoriasis: An observational cohort study. Circulation Research 123(11): 1244-1254. DOI: 10.1161/circresaha.118.313608

Sotos-Prieto, M. & Peñalvo, J.L. 2013. Genetic variation of apolipoproteins, diet and other environmental interactions; An updated review. Nutricion Hospitalaria 28(4): 999-1009. https://www.redalyc.org/articulo.oa?id=309227544004

Srivastava, N., Prakash, J., Srivastava, A., Agarwal, C.G., Pant, D.C. & Mittal, B. 2013. Association of apolipoprotein B XbaI gene polymorphism and lipid profile in northern Indian obese. Indian Journal of Human Genetics 19: 26-31. doi: 10.4103/0971-6866.112880

Tsunoda, K., Harihara, S., Tanabe, Y. & Dashnyam, B. 2012. Polymorphism of the apolipoprotein B gene and association with plasma lipid and lipoprotein levels in the Mongolian Buryat. Biochemical Genetics 50: 249-268. doi:10.1007/s10528-011-9468-y

Xiao, D., Huang, K., Chen, Q., Huang, B., Liu, W., Peng, Y., Chen, M., Huang, D., Zou, T. & Yang, J. 2015. Four Apolipoprotein B gene polymorphisms and the risk for coronary artery disease: A meta-analysis of 47 studies. Genes and Genomics 37: 621-632. doi:10.1007/s13258-015-0292-3

Zhang, P., Gao, J., Pu, C. & Zhang, Y. 2017. Apolipoprotein status in type 2 diabetes mellitus and its complications. Molecular Medicine Reports 16(6): 9279-9286. https://doi.org/10.3892/mmr.2017.7831

*Pengarang untuk surat-menyurat; email: hirasohail@cmhlahore.edu.pk