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

Received: 27 September 2023/Accepted: 20 March 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

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*Corresponding author; email: hirasohail@cmhlahore.edu.pk