Sibiya Thabani, Ghazi Terisha, Mohan Jivanka, Nagiah Savania, Chuturgoon Anil A
Department of Medical Biochemistry and Chemical Pathology, School of Laboratory Medicine and Medical Sciences, College of Health Sciences, University of KwaZulu-Natal, Howard College Campus, Durban 4013, South Africa.
Department of Human Biology, Medical Programme, Faculty of Health Sciences, Nelson Mandela University Missionvale, Bethelsdorp, Port Elizabeth 6059, South Africa.
Plants (Basel). 2022 Dec 26;12(1):119. doi: 10.3390/plants12010119.
The introduction of highly active antiretroviral therapy (HAART) in the treatment of HIV/AIDS has recently gained popularity. In addition, the significant role of microRNA expression in HIV pathogenesis cannot be overlooked; hence the need to explore the mechanisms of microRNA expression in the presence of HAART and Spirulina platensis (SP) in HepG2 cells. This study investigates the biochemical mechanisms of microRNA expression in HepG2 cells in the presence of HAART, SP, and the potential synergistic effect of HAART−SP. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine cell viability following SP treatment. The cellular redox status was assessed using the quantification of intracellular reactive oxygen species (ROS), lipid peroxidation, and a lactate dehydrogenase (LDH) assay. The fluorometric JC-1 assay was used to determine mitochondrial polarisation. The quantitative polymerase chain reaction (qPCR) was also employed for micro-RNA and gene expressions. The results show that MiR-146a (p < 0.0001) and miR-155 (p < 0.0001) levels increased in SP-treated cells. However, only miR-146a (p < 0.0001) in HAART−SP indicated an increase, while miR-155 (p < 0.0001) in HAART−SP treatment indicated a significant decreased expression. Further inflammation analysis revealed that Cox-1 mRNA expression was reduced in SP-treated cells (p = 0.4129). However, Cox-1 expression was significantly increased in HAART−SP-treated cells (p < 0.0001). The investigation revealed that HepG2 cells exposed to HAART−SP treatment showed a significant decrease in Cox-2 (p < 0.0001) expression. mRNA expression also decreased in SP-treated cells (p < 0.0001); therefore, SP potentially controls inflammation by regulating microRNA expressions. Moreover, the positive synergistic effect is indicated by normalised intracellular ROS levels (p < 0.0001) in the HAART−SP treatment. We hereby recommend further investigation on the synergistic roles of SP and HAART in the expression of microRNA with more focus on inflammatory and oxidative pathways.
高效抗逆转录病毒疗法(HAART)在治疗HIV/AIDS方面的应用近来颇受关注。此外,微小RNA表达在HIV发病机制中的重要作用不容忽视;因此,有必要探索在HAART和钝顶螺旋藻(SP)存在的情况下,HepG2细胞中微小RNA表达的机制。本研究调查了在HAART、SP存在的情况下,HepG2细胞中微小RNA表达的生化机制,以及HAART - SP的潜在协同作用。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法测定SP处理后的细胞活力。通过定量细胞内活性氧(ROS)、脂质过氧化以及乳酸脱氢酶(LDH)测定来评估细胞氧化还原状态。使用荧光JC-1法测定线粒体极化。定量聚合酶链反应(qPCR)也用于微小RNA和基因表达分析。结果显示,在SP处理的细胞中,MiR-146a(p < 0.0001)和miR-155(p < 0.0001)水平升高。然而,在HAART - SP处理中,只有MiR-146a(p < 0.0001)呈现升高,而miR-155(p < 0.0001)在HAART - SP处理中表达显著降低。进一步的炎症分析表明,在SP处理的细胞中,Cox-1 mRNA表达降低(p = 0.4129)。然而,在HAART - SP处理的细胞中,Cox-1表达显著升高(p < 0.0001)。研究发现,接受HAART - SP处理的HepG2细胞中Cox-2表达显著降低(p < 0.0001)。在SP处理的细胞中mRNA表达也降低(p < 0.0001);因此SP可能通过调节微小RNA表达来控制炎症。此外,可以通过HAART - SP处理中细胞内ROS水平的归一化(p < 0.0001)来表明其正协同效应。我们在此建议进一步研究SP和HAART在微小RNA表达中的协同作用,更多地关注炎症和氧化途径。