Oxidative Stress and Mitochondrial Dysfunction in Human Immunodeficiency Virus Patients on Highly Active Antiretroviral Therapy

October 18, 2022 ChinonsoIbeh Educational Resources 0

 – Oxidative Stress and Mitochondrial Dysfunction in Human Immunodeficiency Virus Patients on Highly Active Antiretroviral Therapy –

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ABSTRACT

This study was aimed at evaluating oxidative stress and mitochondrial dysfunction in HIV patients on a combination of emtricitabine, tenofovir and efavirenz (ATRIPLA), commonly used for treating these patients attending the HIV clinic of Enugu State University Teaching (ESUT) Hospital, Parklane, Enugu, Nigeria following short and long-term therapy.

Ninety six (96) subjects (divided into four groups) aged between 18 and 60 years were recruited for the study from the HIV clinic and the laboratory of the hospital. Ethical clearance was obtained from the Ethical Committee of the same hospital. Group 1 consisted of twenty four (24) apparently healthy HIV sero-negative age–matched individuals (No HIV or control group) who work in different laboratories of the hospital.

Group 2 consisted of 24 sero-positive patients who had not started any form of treatment (treatment naive group), while group 3 was made up of twenty four (24) subjects on a short-term course of highly active antiretroviral therapy (HAART) (for less than one year).

Group 4 was made up of twenty four (24) subjects who were on HAART for more than one year, representing those on long-term therapy. Ten mililiters (10 ml) of blood sample was collected from each patient from the antecubital-vein without venous stasis into EDTA and plain bottles.

Serum was processed from the retracted whole blood and stored in duplicates in cryobottles at -200C for biochemical analyses while the anticoagulated blood samples were further processed for CD4+ cell count and DNA extraction for genomic studies.

Total antioxidant capacity (TAC in nmol/l), malondialdehyde (MDA in mmol/l), lactate level (in mg/dl), creatine kinase activity (Ck-MB isoform in IU/L), triacylglycerols (TAG) concentrations in mg/dl, CD4+ count in cells/μl, alanine aminotransferase (ALT) activity in IU/L and genomic studies were all done using standard operative procedures.

TABLE OF CONTENTS

Title Page           i

Certification         ii

Dedication      iii

Acknowledgements              iv

Abstract          vi

Table of Contents               vii

List of Figures      xii

List of Tables              xiii

List of Abbreviations       xiv

CHAPTER ONE: INTRODUCTION

  • Origin of HIV 2
  • Classification of HIV 3
  • Structure of HIV 4
    • Envelope` 5
    • Capsid 5
    • HIV Genome 5
  • Replication of HIV 5
  • Epidemiology of HIV/AIDS 9
  • Transmission of HIV 11
  • Main Routes of Transmission in Developing Countries 11
    • Heterosexual Sexual Intercourse 11
  • Homosexual-Spread of HIV 12
  • Mother to Child/Vertical Transmission 12
  • Transmission through Blood and Blood Products 13
  • Contaminated Needles and Sharp Objects 13
  • Immunology of AIDS 14
  • Natural History of HIV Infection 15
  • HIV Infection in Children 16
  • Opportunistic Infection in AIDS 16
  • WHO Case-Definition of Adults with AIDS in Africa 16
  • Prevention of HIV Infection 17
    • Preventing Sexual Transmission of HIV 18
    • Preventing transmission of HIV from mother to child 18
    • Preventing HIV being transmitted by Blood Transfusion 18
    • Preventing Transmission of HIV when Associated with Drug Abuse 19
    • Preventing HIV being transmitted through Contaminated Needles 19
  • The Polymerase Chain Reaction (PCR) 19
  • Treatment of HIV Infection 19
  • Antiretroviral Therapy 20
    • Nucleoside Reverse Transcriptase Inhibitors 20
    • Non-Nucleoside Reverse Transcriptase Inhibitors 22
    • Protease Inhibitors 23
  • Integrase Inhibitors 24
  • Fusion Inhibitors 24
  • Chemokine-Receptor Antagonists 25
  • Highly Active Antiretroviral Therapy Regimen 26
  • The Goals for ARV Treatment 27
  • Complications of Antiretroviral Therapy 27
  • Adverse Clinical Events Associated with HAART 28
  • Drug-Drug Interactions 28
  • Drug Resistance 29
  • Immune Reconstitution Syndrome 29
  • Free Radicals 30
    • Generation of Free Radicals 31
  • Damage Produced by Reactive Oxygen Species 33
    • Lipid Peroxidation 33
      • Clinical Significance of Lipid Peroxidation 35
    • Antioxidants 36
      • Protective Mechanisms of Antioxidants 37
      • Classification of Antioxidants 39
        • Vitamin Antioxidants 39
        • Antioxidant Enzymes 41
        • Antioxidant Metabolites 42
  • Oxidative Stress 43
    • Effect of Oxidative Stress on HIV-Positive Cases 43
    • Oxidative Stress in HIV/AIDS: Effect of Antioxidant Diets 43
  • Cellular Consequences of the ROS/Antioxidant Balance 44
  • How Cells Respond to Increasing Exposure to ROS 45
  • Physiological Conditions that Promote AIDS 47
  • Mitochondria 51
    • Mitochondrial Function 51
    • Mitochondrial DNA 52
    • Mitochondrial Dysfunction 53
    • Mitochondria as Source of Reactive Oxygen Species 55
    • Mitochondria as Targets of ROS 57
    • Regulation of Reactive Oxygen Species Production in Mitochondria 58
  • Justification for the Study 59
  • Aim and Objectives of the Study 59
    • Aim of the Study 59
    • Specific Objectives of the Study 59
  • Specific Research Questions 60

CHAPTER TWO: MATERIALS AND METHODS

  • Location of study 61
  • Chemicals/Reagents/ Samples 61
  • Instruments/Equipment 61
  • Methods 62
    • Experimental Design 62
    • Collection of Blood Samples 62
    • Genomic DNA Extraction and Purification 63
      • Processing Whole Peripheral Blood Down to Guanidium Isothiocyanate Lysate 63
      • Genomic DNA Purification from GITC Lysate 64

2.2.3      Experimental Analysis                                                                                              66

  • Quantification of Mitochondrial DNA Content of Samples 66
  • Real Time Quantitative PCR 68
  • Measurement of Serum Total Antioxidant Capacity 69
  • Measurement of Serum Malondialdehyde 70
  • Determination of Serum Lactate Concentration 72
  • Assay of Serum Alanine Aminotransferase activity 73
  • Determination of Serum Creatine Kinase 74
  • Determination of Serum Triacylglycerols Concentration 75
  • Measurement of the CD4+ Cells in the Subjects 76
  • Statistical Analysis 77

CHAPTER THREE: RESULTS

  • House-keeping (control) genes used as normalizers 78
  • mt-104 Gene across the Different Groups of Subjects 79
  • mt-141 Gene across the Different Groups of Subjects 81
  • mt-108 Gene across the Different Groups of Subjects 83
  • mt-78 Gene across the Different Groups of Subjects 85
  • Total Antioxidant Status across the Different Groups of Subjects 87
  • Malondialdehyde Concentration across the Different Groups of Subjects 89
  • Lactate Concentration across the Different Groups of Subjects 91
  • Alanine Aminotransferase Activity across the Different Groups of Subjects 93
  • Creatine Kinase Activity across the Different Groups of Subjects 95
  • Triacylglycerols concentration across the Different Groups of Subjects 97
  • CD4+ Count across the Different Groups of Subjects 99
  • Correlation of CD4+ Count and other parameters studied 101

CHAPTER FOUR: DISCUSSION      

  • Conclusions 110
  • Recommendations 111

REFERENCES 112

INTRODUCTION

Human Immunodeficiency Virus (HIV) is the cause of the fatal condition called acquired immunodeficiency syndrome (AIDS).

AIDS was first recognized in the United States in 1980- 1981 when homosexual men were found to have unusual infections and tumors suggesting an underlying deficiency in their cell-mediated immunity (Maartens et al., 2014).

Evidence exists  to show that HIV has been present in humans for 15-20 years before then but the exact origin of the virus is not yet known (CDC, 2013).

Over the years, relative decline in morbidity and mortality of human immunodeficiency virus (HIV) infection in some countries has been observed due to the use of a potent combined therapy known as highly active antiretroviral therapies (HAART) (Maartens et al., 2014).

It has led to a decrease in viral load (a measure of the amount of HIV virus in the blood), quantitative and qualitative improvement of immune functions in patients, especially CD4+ T- lymphocytes count, resulting in a decrease of infectious complications and a global clinical improvement (Klatt and Silvestri, 2012).

In spite of the positive effects of HAART on the immune system and on the metabolic alterations during HIV infections, it has been reported that the commonly used drugs zidovudine (AZT), didanosine (ddI) and stavudine (d4T) are toxic to hepatocytes and other tissues (Heil et al., 2010).

REFERENCES

Abduljalil, M.M., Liman, H.M., Umar, R.A. and Abubakar, M.G. (2015). Effect of HIV and HAART on antioxidants markers in HIV positive patients in Sokoto State, Nigeria. International Journal of Scientific and Engineering Research, 6(2): 1069-1074.

Akiibinu, M.O., Adeshiyan, A.A. and Olalekan, A.O. (2012). Micronutrients and markers of oxidative stress in symptomatic HIV-positive/AIDS Nigerians: A call for adjuvant micronutrient therapy. Institute of Integrated Omics and Applied Biotechnology Journal, 3(2): 7–11.

Alfadda, A.A. and Sallam, R.M. (2012). Reactive oxygen species in health and disease. Journal of Biomedical and Biotechnology, 936486.

Alimonti, J.B., Ball, T.B. and Fowks, K.R. (2003). Mechanisms of CD4+ T-lymphocyte cell death in human immunodeficiency virus infection and AIDS. Journal of Genetic Virology, 84(7): 1649-1661.

Aniagolu, M.O., Ezeanyika, L.U.S., Parker, J., Shu, N., Ngwu, A.M. and Ikegwuonu, J. (2015). Changes in some antioxidants in HIV-positive patients on highly active antiretroviral therapy (case study of Nsukka, South East Nigeria). Asian Journal of Science and Technology, 6(4): 1289-1292.

StudentsandScholarship Team.

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