Evaluation of the Immunomodulatory Activity of Hoslundia opposita Vahl (Lamiaceae) Leaf Extract

Filed in Articles by on September 21, 2020

Evaluation of the Immunomodulatory Activity of Hoslundia opposita Vahl (Lamiaceae) Leaf Extract.

ABSTRACT 

The immunomodulatory properties of methanol extract and fractions of Hoslundia opposita leaf on Delayed-Type Hypersensitivity (DTH), primary and secondary humoral response and in vivo leucocyte mobilization were evaluated.

Acute toxicity test of crude extract and phytochemistry study was carried out. The methanol extract (ME) at 200, 400 and 800 mg/kg body weight produced significant (P<0.05) inhibition of DTH response in rat by 85, 75 and 85%.

The n- hexane, ethyl acetate and methanol fractions at 200, 400 and 800 mg/kg body weight produced significant (P<0.05) inhibition of DTH response.

Treatment of rats with single intraperitoneal injection of carrageenan after oral administration of extract and fractions resulted in an increase in leucocyte mobilization into the rat’s peritoneal fluid which was significant.

The total leucocytes counts (TLC) were higher in the extract and fractions treated groups when compared to the control group.

However a fall in primary and secondary antibody titre was observed with both ME and fractions suggesting that Hoslundia opposita may act through a cell mediated mechanism. The crude extract administered (orally) at 5000 mg/kg did  not cause lethality after 48 h observation period.

Phytochemistry tests revealed abundant presence of resins, terpenoids, flavonoids, saponins and glycosides in the fractions and crude extract.

These findings suggest that these may be responsible for the immunomodulatory effect of the leaves of Hoslundia opposita.

It was therefore recommended that further studies be done to identify and isolate the exact constituent responsible for the immunomodulatory effects and also establish mechanism of action of Hoslundia opposita Vahl (Lamiaceae).

TABLE OF CONTENTS

Title page………….. i
Certification……………. ii
Dedication…………… iii
Acknowledgement…………… iv
Abstract……………… vi
Table of content…………… vii
List of tables……………. x
List of figures……………….. xi

Chapter One: Introduction

1.1 Introduction…………………. 1
1.1.1 History of Immunology 1
1.1.1.1 The role of smallpox in the development of vaccination…………. 2
1.1.1.2 Edward Jenner and the development of the first safe vaccine for smallpox…………4
1.1.1.3 Koch, Pasteur and the germ theory of disease…………………….. 4
1.1.2 Immune response………………………………………………….. 5
1.1.2.1 Innate / non-specific immunity………………. 8
1.1.2.1.1 Components of the innate immune system……… 9
1.1.2.1.1.1 Anatomic barriers…………………………………………………. 9
1.1.2.1.1.2 Humoral and chemical barrier………. 10
1.1.2.1.1.2.1 Inflammation……………………… 10
1.1.2.1.1.2.2 Complement system and others………….. 11
1.1.2.1.1.3 Cellular barrier…………………… 13
1.1.2.2 Adaptive/Acquired Immunity…………. 15
1.1.2.2.1 Lymphocytes…………………… 15
1.1.2.2.1.1 Natural Killer cells……………… 15
1.1.2.2.1.2 T cells and B cells………………. 16
1.1.2.2.2 B lymphocytes and antibodies production……… 20
1.1.2.2.3 Alternative adaptive Immune system………… 21
1.1.2.2.4 Immunological memory…………………… 21
1.1.2.2.4.1 Passive memory………………………… 21
1.1.2.2.4.2 Active memory and Immunization……………….. 22
1.1.2.3 Passive Immunity…………………… 22
1.1.2.3.1 Naturally acquired passive immunity……………….. 23
1.1.2.3.2 Artificially acquired passive immunity…………… 24
1.1.2.3.3 Passive transfer of cell-mediated immunity…… 24
1.1.3 Parts of the immune system…………. 25
1.1.3.1 Organs of the immune system………. 25
1.1.3.2 Cells of the immune system………… 30
1.1.4 Pattern recognition receptors…………….. 33
1.1.5 Bystander T-cell apoptosis…………….. 34
1.2 Disorders of human immunity………………. 35
1.2.1 Immunodeficiency…………….. 35
1.2.2 Autoimmunity………………….. 36
1.2.3 Hypersensitivity…………………….. 37
1.3 Immunomodulators……………………. 38
1.3.1 Immunosuppressants…………………… 38
1.3.2 Immunostimulants……………………… 42
1.3.3 Tolerogens……………………… 45
1.3.4 Medicinal Plants with immunomodulatory effect………………… 47
1.4 Botanical Profile of Hoslundia opposita………………………….. 49
1.4.1 Plant taxonomy…………………………. 49
1.4.2 Description of plant…………………………….. 50
1.4.3 Geographical distribution of plant………… 50
1.4.4 Ecology……………………………………. 51
1.4.5 Cultivation of Hoslundia opposita…………… 51
1.4.6 Uses…………………………………….. 51
1.4.7 Ethnomedicinal uses………………. 52
1.5 Literature review……………….. 52
1.6 Aim and scope of study……… 53

Chapter Two: Materials and method

2.1 Materials…………………… 54
2.1.1 Animals………………………. 54
2.1.2 Drugs……………. 54
2.1.3 Chemicals, solvents and reagents………….. 54
2.1.4 Equipments………………… 55
2.2 Methods……………………………. 55
2.2.1 Collection, identification and Preparation of plant material………. 55
2.2.2 Extraction of plant material…………….. 55
2.2.3 Fractionation of methanol extract……………. 56
2.2.4 Determination of extract yield……….. 56
2.2.5 Phytochemical analysis of extract and fractions………………….. 56
2.2.5.1 Test for saponins………………… 56
2.2.5.2 Test for tannins………. 57
2.2.5.3 Test for resins………………………… 57
2.2.5.4 Test for flavonoids………………… 57
2.2.5.5 Test for steroids and terpenoids…………. 57
2.2.5.6 Test for alkaloids………………… 58
2.2.5.7 Test for glycosides………………… 58
2.2.5.8 Test for oil…………………. 59
2.2.5.9 Test for carbohydrate……………… 59
2.2.5.10 Test for reducing sugar…………………. 59
2.3 Pharmacological studies……………………… 59
2.3.1 Acute Toxicity (LD50) of ME……………… 59
2.3.2 Carraageenan induced leucocyte mobilization in rats……………… 60
2.3.3 Sheep Red blood cell-induced delayed type hypersensitivity assay. 60
2.3.4 Hemagglutination antibody titre in rats…………………………… 61
2.4 Statistical analysis…………………. 62

Chapter Three: Results

3.0 Results…………………… 63
3.1 Extraction and fractionation……………….. 63
3.2 Phytochemical constituents of extract and fractions………………. 63
3.3 Acute toxicity (LD50) of ME….. 65
3.4 Pharmacological studies………………… 65
3.4.1 Effect of ME and fractions on in vivo leucocyte mobilization in rat. 65
3.4.2 Effect of ME and fractions on Delayed type hypersensitivity reaction in rats…68
3.4.3 Effect of ME and fractions on Hemaglutination Antibody Titre in Rats…70

Chapter Four: Discussion and Conclusion

4.0 Discussion and Conclusion…………… 72
4.1 Discussion……………… 72
4.2 Conclusion………….. 77
References………………. 78
Appendices………… 94

INTRODUCTION 

1.1 History of Immunology

Immunology is a broad branch of biomedical science that covers the study of all aspects of immune system in all living organisms. It is the branch of medicine and biology concerned with immunity.

Immunity is a biological term that describes a state of having sufficient biological defenses to avoid infection, disease, or other unwanted biological invasion. The concept of immunity has intrigued mankind for thousands of years (Silverstein, 1989).

The prehistoric view of disease was that it was caused by supernatural forces, and that illness was a form of theurgic punishment for “bad deeds” or “evil thoughts” visited upon the soul by the gods or by one’s enemies (Lindquester, 2006).

The ancient historic view was that disease was spontaneously generated instead of being created by microorganisms that grow by reproduction (Madigan and Martinko, 2005).

Between the time of Hippocrates and the 19th century, when the foundations of the scientific methods were laid, diseases were attributed to an alteration or imbalance in one of the four humors (blood, phlegm, yellow bile or black bile), (Silverstein, 1989).

Also popular during this time was the miasma theory, which held that diseases such as cholera or the Black Plague were caused by a miasma, a noxious form of “bad air” (Lindquester, 2006). If someone were exposed to the miasma, they could get the disease.

REFERENCES

Ahsan,N., Hricik, D., Matas, A., Rose, S., Tomlanovich, S., Wilkinson, A., Ewell, M., Mclntosh,M., Stablein, D., Hodge, E. (1999). Prednisone withdrawal in kidney transplant recipients on cyclosporine and mycophenolate mophetil-a prospective randomized study. Steroid withdrawal study group. Transplantation, 68: 1865–1874.

Akah, P.A., Odo, C.I. (2010). Hepatoprotective effects of the solvent fractions of the stem of Hoslundia opposita Vahl. (Lamiaceae) against Carbon tetrachloride and paracetamol- induced liver damage in rats. International Journal of Green Pharmacy, 4(1): 54–58.

Akbar, A.N., Terry, L., Timms, A., Beverly, P.C., Janossy, G. (1988). Loss of CD45R and gain of UCHL1 reactivity is a feature of primed T cells: Journal of Immunology, 140(7): 2171–2178.

Akolade, J.O., Muhammed, N.O., Usman, L.3., Owolarafe, T.A. Oloyede, O.B. (2011). Antidyslipidemic effect of leaf essential oil of Hoslundia opposita Vahl. In Allosan- induced diabetic rats. International Journal of Tropical Medicine and Public Health, 1(1): 1–29.

Alan, M.K., Terry, B.S., Jeffrey, A.B. (2001). Immunomodulators: immunosuppressive agents, tolerogens and immunostimulants. Goodman and Gilman’s The pharmacological basis of therapeutics; 10th ed. United States: McGrawHill Companies.

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