|Year : 2020 | Volume
| Issue : 2 | Page : 168-171
Study of free radicals and antioxidant status in human immunodeficiency virus-positive patients
Ranjit Sidram Ambad1, Rakesh Kumar Jha1, Archana Dhok2, Nandkishor Bankar3
1 Department of Biochemistry, Datta Meghe Medical College, Shalinitai Meghe Hospital and Research Centre, Nagpur, India
2 Department of Biochemistry, Jawaharlal Nehru Medical College, Datta Meghe Institute of Medical Sciences (Deemed to be University), Sawangi (Meghe), Wardha, Maharashtra, India
3 Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Medical Sciences (Deemed to be University), Sawangi (Meghe), Wardha, Maharashtra, India
|Date of Submission||14-Apr-2020|
|Date of Decision||17-Apr-2020|
|Date of Acceptance||22-Apr-2020|
|Date of Web Publication||21-Dec-2020|
Dr. Rakesh Kumar Jha
Department of Biochemistry, Datta Meghe Medical College, Shalinitai Meghe Hospital and Research Centre, Wanadongri, Hingana, Nagpur - 441 110, Maharashtra
Source of Support: None, Conflict of Interest: None
Introduction: Human immunodeficiency virus (HIV) stands for immunodeficiency virus in humans. HIV destroys the immune system of the body, phasing out its capacity to combat diseases and other cancers. HIV can develop into acquired immunodeficiency syndrome (AIDS) when left untreated. It is the severest HIV level and is typically fatal. There is no treatment for HIV at this moment. This can take as long as 10 years to experience more serious symptoms after the infection. A person with <200 cells/mm3 is beginning to develop severe infections called opportunistic diseases and is moving toward AIDS. A free radical can be described as any molecular species capable of independent life in an atomic orbital that contains an unpaired electron. Antioxidants function as a radical scavenger, a donor of hydrogen, a donor of electrons, a decomposer of peroxides, a single quencher of oxygen, an inhibitor of enzymes, a synergist, and a metal chelator. In the intracellular and extracellular environment, both enzymatic and nonenzymatic antioxidants exist for detoxification of reactive oxygen species (ROS). Aim: The aim was to study the free radicals and antioxidant status in HIV-positive patients. Materials and Methods: The present study includes a total of sixty participants that include thirty HIV-positive patients and thirty healthy individuals. Blood samples collected from the participants were obtained for serum glutathione reductase (GSH), superoxide dismutase (SOD), and malondialdehyde (MDA) estimation. Results: The GSH, SOD, and MDA levels in HIV-positive patients were statistically significant. Conclusion: ROS was described as playing a critical role in accelerating and regulating AIDS development. In addition, it was found that antioxidant depletion was a typical sign at the onset of HIV infection, resulting in extreme OS.
Keywords: Antioxidant, free radicals, glutathione reductase, superoxide dismutase and malondialdehyde
|How to cite this article:|
Ambad RS, Jha RK, Dhok A, Bankar N. Study of free radicals and antioxidant status in human immunodeficiency virus-positive patients. J Datta Meghe Inst Med Sci Univ 2020;15:168-71
|How to cite this URL:|
Ambad RS, Jha RK, Dhok A, Bankar N. Study of free radicals and antioxidant status in human immunodeficiency virus-positive patients. J Datta Meghe Inst Med Sci Univ [serial online] 2020 [cited 2021 Jan 26];15:168-71. Available from: http://www.journaldmims.com/text.asp?2020/15/2/168/304234
| Introduction|| |
When the first incident of human immunodeficiency virus (HIV) infection was identified in 1981, clinicians and researchers were much concerned about its effects and symptoms. Its distribution around the world posed a serious issue. The resulting opportunistic infectious diseases and slow progression to acquired immunodeficiency syndrome (AIDS) are troubling. Novel advancements in treatment and therapy brought researchers much hope. Researchers claimed that if the infection could be catch early and the agents that contributed to its life-threatening effects, medication might alter the path of these agents and save lives. However, as most virologists realized, it would not be an easy mission.
In HIV patients, high oxidative stress was associated with infection with the HIV and antiretroviral therapy. In HIV patients, the difference in antioxidant-oxidant rates favors viral replication and progression of disease.
Retroviruses are RNA-containing viruses that replicate by virtue of a viral RNA-dependent DNA polymerase, also called reverse transcriptase, through an intermediate DNA. The retroviridae family is composed of three subfamilies. Oncovirinae contains all the oncogenic retroviruses and other nononcogenic viruses which are closely related. Lentivirinae includes “soft” viruses such as visna-maedi virus, caprine arthritis encephalitis virus, equine infectious anemia virus, and immunodeficiency viruses, most notably HIV, which is the causative agent of AIDS.
Following its discovery, the HIV remains a significant problem for the health-care delivery system around the world. There were many attempts to find a solution, but none could see the light of day. The presence of this virus in the body system puts a burden on metabolism in the body. It produces free radicals and affects cellular organelles, which can exacerbate the effect that HIV has on the body system.
Global mortality and morbidity of human immunodeficiency virus/acquired immunodeficiency syndrome
After the first case was identified in 1981, the HIV/AIDS epidemic has undergone drastic changes. HIV transmission was originally regarded as a disease among gay men or Haitians in Western countries and has been recorded in nearly every part of the world. For several sub-Saharan African cities, the prevalence rates reached more than 30% among adults in the 1990s, and no free, successful care was available.
Reactive oxygen species (ROS), metabolic by-products in both prokaryotic and eukaryotic cells, is thought to be harmful to the host by destroying cellular macromolecules including DNA, proteins, and lipids, contributing to disease pathogenesis. It has been well recognized, however, that ROS is essential for the maintenance of host physiological functions and plays an important role in infection control. Since the late 80s, ROS association with HIV pathogenesis and disease progression has been hypothesized. Numerous clinical trials were performed (reviewed in) on regulation of ROS in HIV-infected patients, but the findings were not reliable. Bearing in mind the dynamic role of ROS in infection and immune response, understanding the balance between redox biology and oxidative stress can provide insights into infection and disease prevention and treatment.,
In HIV-infected patients, the increase of the concentration of free radicals is related to: a depletion of protective system (glutathione reductase [GSH] peroxidase, superoxide dismutase [SOD], Vitamin E, and selenium) and an increased production of free radicals (superoxide anion, hydrogen peroxide, and hydroxyl radical) consecutive to the activation of lymphocytes and phagocyting cells, the chronic inflammation, the increased polyunsaturated fatty acid concentration and lipoperoxidation, and direct or indirect effect of several pathologic agents including Mycoplasma sp. This free radical excess could disrupt cell membranes and cause apoptosis, which is the main cause of CD4+ depletion lymphocytes. The role of free radicals in the pathogenesis of HIV infection is discussed with regard to data published in the literature after a brief analysis of the free radicals' synthesis pathway, their possible deleterious effects, and the defense systems.
Role of free radicals in human immunodeficiency virus
Within a few years of HIV discovery, oxidative stress was first found in the HIV pathology. For HIV-infected patients, GSH is reduced for serum, lymphocyte, monocyte, and lung epithelial lining.,, GSH depletion in plasma of HIV-positive patients is inversely associated with increased GSSG (GSH disulfide), indicating that free radical defenses have been weakened. Thioredoxin is reduced in the lymph node dendritic cells but elevated in the plasma of patients with HIV infection.
An antioxidant is a molecule that is stable enough to contribute and neutralize an electron to a rampaging free radical, thus reducing its ability to damage. These antioxidants mainly delay or inhibit cell damage through their free radical scavenging properties. These low-molecular-weight antioxidants can interact safely with free radicals and terminate the chain reaction before destroying essential molecules. Any of those antioxidants, including GSH, ubiquinol, and uric acid, occur during normal body metabolism.
Mechanism of action of antioxidant
Two major mechanisms of action for antioxidants have been proposed. The first is a chain-breaking mechanism by which the primary antioxidant donates an electron to the free radical present in the systems. The second mechanism involves eliminating initiators of the ROS/reactive nitrogen species (secondary antioxidants) by quenching the catalyst that initiates the chain. Antioxidants can exert their effect on biological systems through various mechanisms, including donation of electrons, chelation of metal ions, co-antioxidants, or by regulation of gene expression.
Role of antioxidants
Importantly, weak antioxidant Vitamins C and E that may lead to some manifestations. Antioxidants such as Vitamins E and A, as well as Vitamin C, and GSH inhibit free radical damage in the cytosol. Such enzymatic systems also help inactivate free radical reactions. Hydrogen peroxide and superoxide anion are broken down by these. The enzymes appear to be located near the oxidant generation sites. Catalase present in peroxisomes decomposes H2O2 (2H2O2 = O2 +2H2O) hydrogen peroxide. SODs present in many types of cells bind superoxides to inactivate molecules, H2O2 (2O2 -+2H). Reportedly, this group contains both the manganese-SOD present in the cytosol. Through detoxifying oxygen-free radicals, GSH also offers defense against cellular injury.
| Materials and Methods|| |
The study was conducted in the Department of Biochemistry and Department of Medicine at Datta Meghe Medical College, Shalinitai Meghe Hospital and Research Centre, Hingana Nagpur, in collaboration with Jawaharlal Nehru Medical College and AVBRH (Datta Meghe Institute of Medical sciences) Sawangi (Meghe) Wardha, Maharashtra.
- Group I: Thirty HIV-positive patients from group 1 were enrolled from the medicine ward
- Group II: Thirty healthy individuals from the normal population.
A total of sixty participants were enrolled and were grouped as mentioned ahead.
- Hospitalization in the past 3 months
- Untreated hypothyroidism or hyperthyroidism
- Known diabetes mellitus, hypercortisolemia, and coronary artery disease
- Known liver impairment (ALT and AST >2 ULN)
- Renal impairment (creatinine >1.4)
- Inability to walk
- Patients on anticoagulation or antiplatelet therapy.
This was a cross-sectional study.
Five milliliters of venous blood was collected under strict aseptic conditions in plain bulb for serum. Serum was used for the estimation of GSH, SOD, and malondialdehyde (MDA).
Serum antioxidants (GSH/SOD) were analyzed by ELISA method., Serum free radical (MDA) was analyzed by— measured by using UV-VIS spectroscopy.
Data collected were entered into Microsoft Excel worksheet and statistically analyzed using the Statistical Package for the Social Sciences version 20 (Chicago, Illinois, USA). For quantitative data, mean, standard mean, standard deviation, t-test, and Karl Pearson's coefficient of correlation were calculated. P <0.05 (0.01) was considered as statically significant (highly significant) at 95% confidence interval.
Ethical approval for this study (DMIMS(DU)/IEC/2019-20/3002) was provided by the Ethical Committee of Datta Meghe Institute of Medical Sciences (Deemed to be University) on 22nd July 2019.
| Results|| |
[Table 1] shows that the GSH level in HIV-positive patients was 3.09 ± 0.95 as compared to control group 4.17 ± 0.80, which is statistically significant. The level of SOD in HIV patients was 31.56 ± 9.47 and in control group 19.81 ± 8.14, serum MDA level in HIV patients were 5.68 ± 0.89 and in control group level of MDA were 3.45 ± 0.68 which is statistically significant.
|Table 1: Comparison of mean glutathione reductase, superoxide dismutase, and malondialdehyde in human immunodeficiency virus patients and control group|
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| Discussion|| |
Our study shows that the GSH level in HIV-positive patients was 3.09 ± 0.95 as compared to control group 4.17 ± 0.80, which is statistically significant. Pathak and Shetty and Rajekar and Ambad have shown that the development of plasma GSH peroxidase and SOD is gradually decreasing as pregnancy progresses in HIV.,
Pasupathi et al. and Rajekar and Ambad and has been shown that HIV-infected individuals have decreased antioxidant concentrations, impaired GSH metabolism and increased spontaneous ROS generation. In addition, there has been recorded a major decrease in GR levels. To be cognizant, it is understood that lower GR levels and overall oxidative stress regulate inflammatory cytokine activity., Another study done by Rajekar and Ambad showed that the GSH level was decreased in HIV pregnant women compared to normal pregnant women.
[Table 1] shows that the level of SOD in HIV patients was 31.56 ± 9.47 and in control group 19.81 ± 8.14, which is statically significant; our study is supported by Gil L et al. and Rajekar and Ambad According to CDC 1993 recommendations, patients were split into two categories. Increases in CD95 and CD38 mirror the severity of the HIV infection. The plasma of HIV + patients observed both a decrease in GSH levels and an increase in MDA and TH levels. Such patients also reported an increase in lymphocyte DNA fragmentation as well as a large reduction in GPx (P < 0.05) and an increase in erythrocyte SOD activity. HIV-infected patients had major variations in global indexes of total antioxidant status compared to the control community. The reduced total antioxidant capacity may be attributed to depleted antioxidant defense mechanisms.,
[Table 1] shows that serum MDA level in HIV patients was 5.68 ± 0.89 and in control group, the level of MDA was 3.45 ± 0.68, which is statistically significant; our study is supported by Awodele et al. and Rajekar and Ambad. In the HIV-TB co-infection population, a high MDA value was identified relative to HIV patients on HAART, and this increase was due to increased ROS in HIV and TB infections. Salisu et al. and Rajekar and Ambad showed elevated levels of MDA plasma, as well as a very elevated index of lipid peroxidation in co-infection with HIV/TB. No gender gap was found in the MDA principles.,,,,
| Conclusion|| |
ROS was described as playing a critical role in accelerating and regulating AIDS development. In addition, it was found that antioxidant depletion was a typical sign at the onset of HIV infection, resulting in extreme OS. This discrepancy gave way to the pro-activating the virus replication and transcription pathways.
Hence, antioxidant supplementation was thought to be able to counteract the symptoms of OS and delay the progression to AIDS. Although some findings are positive for their therapeutic use for HIV infection, study supported by Rajekar and Ambad showed similar conclusion, further studies are needed to validate these results. There are still limitations about the stage of infection, the proper dosage and length of antioxidants, and the resistance to treatment. Randomized trials using adequate antioxidant(s) dosages are important to investigate their role in changing the course of HIV infection.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Quaye O, Kuleape JA, Bonney EY, Puplampu P, Tagoe EA. Imbalance of antioxidant enzymes activities and trace elements levels in Ghanaian HIV-infected patients. PLoS One 2019;14:e0220181.
Yashwantrai N. Vaishnav and Flossie Wong-Staal the biochemistry of aids Annu. Rev Biochem 1991;60:577-630.
Rajekar RB, Ambad RS. Int J Med Sci Innovative Res 2017;2:330-7.
Rajekar RB, Ambad RS., Global Mortality and Morbidity of HIV/AIDS, Major Infectious Diseases. Int J Med Sci Innovative Res 2017;2:330-7.
Liu T, Stern A, Roberts LJ. The isoprostanes: Novel prostanglandin-like products of the free radical catalyzed peroxidation of arachidonic acid. J Biomed Sci 1999;6:226-35.
Rabaud C, Tronel H, Fremont S, May T, Canton P, Nicolas JP. Free radicals during HIV. Infect Ann de Biologie Clinique 1997:55:565-71.
Eck HP, Gmünder H, Hartmann M, Petzoldt D, Daniel V, Dröge W. Low concentrations of acid-soluble thiol (cysteine) in the blood plasma of HIV-1-infected patients. Biol Chem Hoppe Seyler 1989;370:101-8.
De Quay B. Glutathione reductase depletion in HIV-infected patients: Role of cysteine deficiency and effect of oral N-acetylcysteine. AIDS 1992;6:815-20.
López Galera RM, Juárez Giménez CJ, Montoro Ronsano JB, Segura Cardona RM, Arbós Via MA, AltisentRoca C, et al
. Glutathione reductase and cysteine in HIV-infected hemophiliacs. Clinica Chimica Acta 1996;254:63-72.
Nakamura H, De Rosa S, Roederer M, Anderson MT, Gregson Dubs J, Yodoi J, et al
. Elevation of plasma thioredoxin levels in HIV-infected individuals. Int Immunol 1996;8:603-11.
Halliwell B. How to characterize an antioxidant-An update. Biochem Soc Symp 1995;61:73-101.
Shi HL, Noguchi N, Niki N. Comparative study on dynamics of antioxidative action of α- tocopheryl hydroquinone, ubiquinol and α- tocopherol, against lipid peroxidation. Free RadicBiol Med 1999;27:334-46.
Rice-Evans CA, Diplock AT. Current status of antioxidant therapy. Free Radic Biol Med 1993;15:77-96.
Krinsky NI. Mechanism of action of biological antioxidants. Proc Soc Exp Biol Med 1992;200:248-54.
Wasnik RR, Akarte NR. Evaluation of serum zinc and antioxidant vitamins in adolescent homozygous sickle cell patients in Wardha, district of central India. J Clin Diagn Res 2017;11:BC01-3.
Uloaku Akunueze EN, Ifeanyi OE, Onyemobi EC, Johnson N, Chijioke Uzoanya EA. Antioxidants in the management of human immunodeficiency virus infection. J HIV Retro Virus 2018;4:1-2.
Goldberg DM, Spooner RJ. Assay of Glutathione Reductase. In: Bergmeyen, H.V., Ed., Methods of Enzymatic Analysis, 3rd Edition, Vol. 3, Verlog Chemie, Deerfiled Beach, 258-265.; 1983. p. 258-65.
Miller NJ, Rice-Evans C, Davies MJ, Gopinathan V, Milner A. A novel method for measuring antioxidant capacity and its application to monitoring the antioxidant status in premature neonates. Clin Sci 1993;84:407-12.
Weitner T, SuzanaInić I, Jablan J, Gabričević M, Domijan AM. Spectrophotometric determination of malondialdehyde in urine suitable for epidemiological studies. Croat Chem Acta 2016;89:133-9.
Pathak SS, Shetty DN. Essentially zinc in pregnancy to maintain antioxidant status. Indian Practitioner 2001;54:766-70.
Pasupathi P, Ramachandran T, Sindhu PJ, Saravanan G. Enhanced oxidative stress markers and antioxidant imbalance in HIV infection and AIDS patients. J Sci Res 2009;1:370-80.
Gil L1, Martínez G, González I. Contribution to characterization of oxidative stress in HIV/AIDS patients. Pharmacol Res 2003;47:217-24.
Baliga S, Chaudhary M, Bhat SS, Bhatiya P, Thosar N, Bhansali P. Determination of toral antioxidant capacity of saliva in sickle cell anemic patients-A cross-sectional study. J Indian Soc Pedod Prev Dent 2017;35:14-8.
] [Full text]
Awodele O, Olayemi SO, Nwite JA, Adeyemo TA. Investigation of the levels of oxidative stress parameters in HIV and HIV-TB co-infected patients. J Infect Dev Ctries 2012;6:79-85.
Salisu AI, Dissi GM, Muhammad S, Ahmad S, Waziri BI. Serum Vitamin E And Malondialdehyde in Hiv/Tb Co-Infected Patients In Kano State, Nigeria. BJMLS 2018;3:117-25.
Wasnik R, Akarte NR. Evaluation of Serum Zinc and Antioxidant Vitamins in Adolescent Homozygous Sickle Cell Patients in Wardha, District of Central India. J Clin Diagn Res 2017;11:BC1-3. Available from: https://doi.org/10.7860/JCDR/2017/30855.10320.
[Last accessed on 2020 Feb 11].
Gupta RK, Deogade MS. Antioxidant Activities of Martynia Annua Linn. Root Extract. Int J Ayurveda Med 2019;10:326-8.
Kute PK, Muddeshwar MG, Sonare AR. Pro-Oxidant and Anti-Oxidant Status in Patients of Psoriasis with Relation to Smoking and Alcoholism. J Evol Med Dent Sci 2019;8:2677-80. Available from: https://doi.org/10.14260/jemds/2019/582.
[Last accessed on 2020 Feb 11].