The Effect of Ocimum Tenuiflorum (Nchuanwu) Leaf Extract on Hematolgical Parameters of Immumnosuppressed Albino Rats.
ABSTRACT
The effect of Ocimum tenuiflorum on hematological parameters in immunosuppressed albino rats was investigated in this study. The aqueous (AE) and methanol extract (ME) of the leaf were obtained. Acute toxicity study was done to determine the LD50 of the leaf extract.
Rats of mixed sexes, aged 2- 3months, weighing 150 to 240 grams were used. The rats were divided into 8 groups A-H of four rats each. Group A served as normal control. Immune- suppression was induced using 3mg/kg bodyweight of cyclophosphamide (CP) to rats in groups B to F for 5 days. Group B was CP control.
Group C, D, E and F received 500 mg/kg bodyweight (b.wt) of AE, 1000mg/kg b.wt of AE, 500 mg/kg b.wt of ME, and 1000 mg/kg b.wt of ME once daily for 21 days by oral route, using oral cannula.
Groups G & H (none induced) received 500 mg/kg b.wt AE and 1000 mg/kg b.wt ME once daily for 21 days. Rats in all the groups received rat feed and water ad libitum for 21 days.
On days 15 and 22, 2ml of blood were drawn from each rat via the median canthus into EDTA (ethylenediamin tetra acetic acid) bottle. The HB, PCV, total and differential WBC, were determined using automatic cell counter.
Group B CP control of day 15 showed significantly deceased value in HB; 5.8 ± 0.30g/dl, PCV; 17.6 ± 0.74%, WBC; 0.63 ± 0.2×103/µl Neut; 0.75 ± 1.50%, Lym; 3.5 ± 4.73%, compared with the normal control group A: HB; 13.65 ± 0.47g/dl, PCV;41.15 ± 1.35%, WBC; 11.35 ± 1.76 x103/µl, Neut; 22.25 ± 6.55%, Lym; 73.75 ± 6.75, Mono; 2.5 ± 1.73, Eos; 1.75 ± 1.26, (P<0.001).
Groups C, D, E and F, of day 15 had significant increase in (Hb); 11.27 ± 0.85g/dl, 12 ± 0.30g/dl, 11.67 ± 0.38g/dl, 12.45 ± 0.31g/dl when compared with CP control group B: HB value; 5.88 ± 0.30g/dl (P<0.05). Groups C, D, E and F had significantly increased PCV; 33.47 ± 2.17%, 36.53 ± 0.50%, 35,5 ± 1.67%, 37.35 ± 1.12%, when compared with CP control group B; PCV value; 17.58 ± 0.74%, (P<0.01).
(WBC) count on day 15 were increased in groups C, D, E and F, values were; 23.87 ± 24.16 x 103µ/l, 12.07± 2.8 x 103µ/l, 14.97 ± 4.24 x 103µ/l, and 14.3 ± 7.40 x 103µ/l compared with the CP control B; 0.63 ± 0.21 x 103µ/l,( P<0.001).
Lym count was significantly raised in groups C; 30 ± 14.73%, D; 42 ± 14.18%, E; 54.67 ± 8.08%, F; 21.75 ± 11.21%, when compared with the CP control group B; 3.5 ± 4.73%, (P<0.001) Neutrophil count revealed very high count in groups C; 63.33 ± 10.60%, D; 54.33 ± 15.70%, E; 38.33±6.66%, F; 77.75±11.00%, when compared with the CP control 0.75±1.50% (P<0.001).
On day 22, there was an increase in neutrophil count in groups D; 60±14.40%, and F; 52%, when compared with group B; 38±4.24% (p<0.001). Lymphocytes count were significantly increased in groups C; 55.5±12.02%, and E; 60±7.07%, when compared with the CP control group B; 54.64±7.57%, (P<0.05).
Monocyte count was significantly raised in groups C; 2±0.00%, D; 1.15±2.12%, and F; 2%, when compared with control group B; 1±1.00%, (P<0.05) Eosinophil count, were significantly high in groups C; 5±7.07% and E; 4±1.41%, when compared with the CP control group B; 1.33±1.53%, (P<0.001).
The HB concentration revealed significant increase in groups C; 11.65±0.21g/dl, D; 13.15±0.49g/dl, E; 12.25±9.07g/dl F; 13.43±1.33g/dl when compared with the CP control group B; 5.633±0.85g/dl, (P<0.05). The pack cell volume showed significantly increased value in groups C; 35.2±0.85%, D; 39.85±1.34%, E; 36.95±0.21%, F; 41.9%,when compared with the CP control B; 17.43±2.23%, (P<0.01).
The phytochemical analysis of the aqueous and methanol extract of Ocimum tenuiflorum reveal the presence of flavonoids, terpenoids, alkaloids, carbohydrates, proteins, reducing sugar, glycosides, saponin, tannin, protein, steroids, oil, resin, and acidic compound.
The histological analysis of the liver and spleen of the normal control showed normal features of the hepatic and splenic tissues.
The treated group and none induced that received 500mg/kg AE and 1000mg/kg ME showed normal features of the two organs, with mild histopathological changes, which could still be the effect of CP. Ocimum tenuiflorum leaf of aqueous and methanol extract has mild effect on the liver and spleen of albino rats at dose used in this study; hence it is safe for use in traditional medicine.
In conclusion this study has revealed that Ocimum t. extract has the ability to boost suppressed immunity.
TABLE OF CONTENTS
Title Page – – – – – – – – -i
Certification – – – – – – – – – -ii
Dedication – – – – – – – – – – -iii
Acknowledgement- – – – – – – – -iv
Abstract – – – – – – – – – – -x
Table of Contents – – – – – – – – -iv
List of Tables- – – – – – – – – -vi
List of Plates- – – – – – – – – -vii
CHAPTER ONE: INTRODUCTION- – – – – -1
Background of Study- – – – – – – – -1
Aims and Objectives- – – – – – – – -4
CHAPTER TWO: LITERATURE REVIEW- – – – – -6
2.1 Immunosuppression- – – – – – – – -6
2.2 Deliberately induced- – – – – – – – -6
2.3 Cyclophosphamide- – – – – – – – -8
2.4 Blood- – – – – – – – – – -15
2.5 Ocimum tenuiflorum (Nchuanwu) leaf- – – – – – -21
2.6 Active compound of Ocimum tenuiflorum- – – – – -22
2.7 medicinal uses of Ocimum tenuiflorum- – – – – – -23
2.8 digestive system- – – – – – – – -23
2.9 wound healing- – – – – – – – -24
2.10 cardiovascular circulatory system- – – – – – -25
2.11 immune system- – – – – – – – -25
2.12 Skin- – – – – – – – – – -26
2.13 Nervous system- – – – – – – – -28
2.14 Reproductive system- – – – – – – -28
2.15 Respiratory system- – – – – – – – -28
2.16 Urinary system- – – – – – – – -29
2.17 Psycho-spiritual- – – – – – – – -29
CHAPTER THREE: MATERIALS AND METHODS- – – -31
3.1 Collection of plant material- – – – – – -31
3.2 Sample preparation- – – – – – – – -31
3.3 Extraction of sample materials- – – – – – -31
3.4 Aqueous extract- – – – – – – – -31
3.5 Methanol extract- – – – – – – – -31
3.6 Acute toxicity studies- – – – – – – -32
3.7 Animal housing- – – – – – – – -32
3.8 Experimental design- – – – – – – – -33
3.9 Blood sample collection- – – – – – – -34
3.10 Blood sample analysis- – – – – – – -34
3.11 Statistics- – – – – – – – – -35
3.12 Phytochemical analysis of Ocimum tenuiflorum leaf– – – -35
3.12.1 Analysis of crude powder of Ocimum- – – – – -36
3.12.2 Analysis of methanol leaf extract of Ocimum- – – – -40
3.12.3 Analysis of aqueous leaf extract of Ocimum- – – – -45
CHAPTER FOUR: RESULTS- – – – – – -50
CHAPTER FIVE: DISCUSSION AND CONCLUSION- – – -71
5.1 Discussion- – – – – – – – – -71
5.2 Conclusion- – – – – – – – – -76
References- – – – – – – – – – -77
INTRODUCTION
1.1 Background of study
Immunosuppression is the suppression of the immune system and its ability to fight infections. It involves an act that reduces the efficacy of the immune response (Sigal and Dumont, 1992).
Immunosuppression may result from certain diseases such as AIDS or lymphoma, lupus or from certain drugs, used to treat cancer (for example cyclophosphamide).
Immunosuppression may occur as an adverse reaction to treatment of auto-immune diseases like rheumatoid arthritis or Crohn’s disease (Abbas, et al, 2010).
It may be deliberately induced with drugs, to prevent the body from rejecting an organ or treating graft –versus host disease after bone marrow transplant.
Drugs such as steroids, azathioprine, ciclosporin monoclonal antibodies which include basiliximab, zenapax, muromonab, and prednisone are used in organ transplant (kidney, liver, heart) these drugs are also known as immunosuppressant drugs, they inactivates immune system to prevent tissue rejection (Halloran, 2004).
Immunosupprression may occur as a result of radiotherapy. Non-deliberate immunosuppression can occur in malnutrition, aging, many types of cancer such as leukemia, lymphoma, multiple myeloma and certain chronic infections such as Human Immunodeficiency Virus (HIV) which results in increased susceptibility to pathogens such as bacteria and virus (Abbas et al 2010).
Immunosuppression was deliberately induced in this research to evaluate the effect of Ocimum tenuiflorum leaf extract on immune system.
REFERENCES
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