ELUCIDATING THE miR-155-5p AND miR-181a-2-3p AS CIRCULATING MICRORNA BIOMARKERS OF THE INFLAMMATORY REGULATION IN THE PREMATURE ATHEROSCLEROSIS

Christianus Rustin; Muhammad Irfan Sazali  Kamaruddin; Lilik  Herawati; Azrina  Zainal Abidin; Zulhabri Othman

Christianus Rustin School of Graduate Studies, Management and Science University, 40100 Shah Alam, Selangor, Malaysia
Muhammad Irfan Sazali Kamaruddin School of Graduate Studies, Management and Science University, 40100 Shah Alam, Selangor, Malaysia
Lilik Herawati Faculty of Medicine, Universitas Airlangga, 60131 Surabaya, Indonesia
Azrina Zainal Abidin Faculty of Health and Life Sciences, Management and Science University, 40100 Shah Alam, Selangor, Malaysia
Zulhabri Othman Faculty of Medicine, Manipal University College Malaysia, 75150 Melaka, Malaysia.

Keywords: Atherosclerosis, biomarkers, inflammatory, microRNA, miR-155-5p, miR-181a-2-3p

Abstract

MicroRNAs, short fragments of non-coding RNA, play a crucial role in gene regulation found within atherosclerotic plaque complex via inflammatory pathways, indicating their role as a biomarker for atherosclerosis. We hypothesize that miR-155-5p and miR-181a-2-3p are overexpressed in High-Fat Diet (HFD=5) compared to Normal Diet (ND=5) in Sprague Dawley (SD) rats. MicroRNAs quantification was confirmed by Real-Time PCR using SYBR Green within two timelines. The lipid profiling was measured using mission 3-in-1 cholesterol meter, and histology analysis of the aortas was performed using Haematoxylin & Eosin (H&E) method. Lipid profiling analysis revealed as significant higher (p<0.05) of low-density lipoprotein (LDL) (20.3±6.12) mg/dL, triglycerides (TG) (134.65±7.41) mg/dL and very low-density lipoprotein (VLDL) (21.54±8.23) mg/dL in HFD at week 12. Whilst the high-density lipoprotein (HDL) and total cholesterol (TC) were recorded to be significantly higher (p<0.05) in ND groups with 68.62±6.54 mg/dL and 54.09±8.40 mg/dL respectively. The microRNA analysis showed a significant difference (p=0.04; t=7.09) in the microRNA expression between HFD and ND groups. The average Ct value of rno-miR-181a-2-3p and rno-miR-155-5p in HFD group at week 12 were 27.08±10.84 and 25.96±11.27 respectively. The rno-miR-181a-2-3p showed higher average fold changes expression level compared to rno-miR-155-5p. Moreover, an obvious thickening of tunica intima of the aorta due to inflammation activities in HFD (152.50±7.52) µm compared to ND (148.41±1.94) µm. In summary, microRNAs were significantly expressed in HFD compared to ND and supported by the lipids level and aortas inflammation.