Antiulcerogenic and Immunostimulatory Properties of the Aqueous Extract of the Leaves of Ficus Capensis in Wistar Albino Rats
– Antiulcerogenic and Immunostimulatory Properties of the Aqueous Extract of the Leaves of Ficus Capensis in Wistar Albino Rats –
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ABSTRACT
Ficuscapensis leaves have always played a great role in preventing, controlling and alleviating various disease conditions. This study was aimed at validating the traditional use of the leaves of Ficuscapensisin folk medicine as an anti- ulcer and immunostimulatory agent.
Diclofenac sodium (100mg/kg) body weight was administered intraperitoneally to induce ulcer. Seven groups(five rats each)were used. Groups 2(150mg ranithidine),4,5,6and7 served as the ulcer test groups while group1 and 3 served as ulcer untreated group(negative and positive controls respectively).
After 24 hour fasting, all the rats were sacrificed and the stomach was removed for observation of ulcer score, ulcer index and ulcer indices(SOD,CAT, MDA and GSH).Five groups(five rats each) were used to investigate delayed type hypersensitivity(DTH),humoral antibody(HA)and Myelosuppression.
Groups 2,3, 4 and 5 (Levamisol) served as the test groups while group1 was the negative control.DTH in rats was sensitized by subcutaneous (SC) injection of 0.1ml of 109 cells /ml sheep red blood cells (SRBCs) (day 0) in the planter region of the right hind foot paw and was challenged on day 5 by SC injection of the same concentration of antigen into the left hind paw.
The oedema produced by antigenic challenge in the left hind paw was measured with a micrometer screw guage. HA in rats was immunized by ip injection of 0.2ml of 109SRBCs/ml on day 0 and was challenged by injecting the same concentration on day 7.
Primary antibody titre was determined on day 7(before the challenge) and the secondary titre was determined on day 14 by the haemagglutination technique. The highest dilution showing visible haemagglutination was taken as the antibody titre.
TABLE OF CONTENTS
PAGE
Title Page – – – – – – – – – – i
Certification – – – – – – – – – – ii
Dedication – – – – – – – – – – iii
Acknowledgement – – – – – – – – – iv
Abstract – – – – – – – – – – v
Table of Content – – – – – – – – – vi
List of Figures – – – – – – – – – – xiii
List of Tables – – – – – – – – – – xv
CHAPTER ONE: INTRODUCTION
1.1 Definition of ulcer – – – – – – – – 1
1.1.1 Brief history of peptic ulcer diseases – – – – – 1
1.1.2 Pathogenesis of peptic ulcer – – – – – – 2
1.2 Factors that produce ulceration – – – – – – 3
1.2.1 Endogenous factors producing ulceration – – – – – 3
1.2.1.1 Acetylcholine – – – – – – – – 3
1.2.1.2 Gastrin – – – – – – – – – 4
1.2.1.3 Histamine – – – – – – – – – 5
1.2.1.4 Somatostatin and Cholecystokinin – – – – – – 6
1.2.1.5 Calcium ion as a second messenger – – – – – 6
1.2.1.6 Genetic factors – – – – – – – – 6
1.2.2 Exogenous factors producing ulceration – – – – – 6
1.2.2.1 Helicobacter pyroli – – – – – – – – 6
1.2.2.2 Non-steroidal anti-inflammatory drugs – – – – – 8
1.2.2.3 Ethanol – – – – – – – – – 10
1.2.2.4 Cigarette smoking – – – – – – – – 10
1.2.2.5 Diet – – – – – – – – – – 10
1.2.2.6 Psychological factors (Stress ulcers) – – – – – 11
1.2.3 Free radicals/ reactive oxygen species – – – – – 12
1.3 Diagnosis of ulcer – – – – – – – – 14
1.3.1 Endoscopy and Radiology approaches to the diagnosis of ulcer – – 14
1.3.1.1 Culture – – – – – – – – 14
1.3.1.2 Histological assessment – – – – – – – 15
1.3.1.3 Rapid urease tests – – – – – – – – 15
1.3.1.4 Polymerase chain reaction – – – – – – – 15
1.3.2 Non-endoscopic diagnosis of ulcer – – – – – 15
1.3.2.1 Antibody test – – – – – – – – 15
1.3.2.2 Urea breath test – – – – – – – – 16
1.3.2.3 Stool antigen test – – – – – – – – 16
1.4 Therapy for peptic ulcer – – – – – – – 16
1.4.1 Non-pharmacological treatment of peptic ulcer disease – – – 16
1.4.2 Pharmacological Treatment of peptic ulcer disease – – – 17
1.5 An overview of the immune system – – – – – 18
1.5.1 Immunomodulation – – – – – – – 18
1.5.1.1 Immunostimulation – – – – – – – 18
1.5.1.2 Immunosuppression – – – – – – – 19
1.6 Humoral immunity – – – – – – – – 20
1.7 Cell-mediated immunity – – – – – – – 21
1.8 Anaemia – – – – – – – – – 22
1.8.1 Types of anaemia – – – – – – – – 23
1.8.1.1 Iron deficiency anaemia – – – – – – – 23
1.8.1.2 Haemolyticanaemia – – – – – – – 23
1.8.1.3 Acute haemorrhagicanaemia – – – – – – 23
1.8.1.4 Chronic haemorrhagic anemia – – – – – – 24
1.8.1.5 Perniciousanaemia – – – – – – – – 24
1.8.1.6 Aplastic anaemia – – – – – – – – 24
1.9 Haematological indices – – – – – – – 25
1.9.1 Red blood cells (Erythrocytes) – – – – – – 25
1.9.2 Packed Cell Volume (PCV) – – – – – – 25
1.9.3 Haemoglobin – – – – – – – – 25
1.9.4 Total white blood cells (WBCs) count – – – – – 26
1.10 Haematopoiesis – – – – – – – – 26
1.10.1 Vitamin B12- – – – – – – – – 27
1.10.2 Folic acid – – – – – – – – – 27
1.10.3 Vitamin C (Ascorbic Acid) – – – – – – 28
1.10.4 Iron – – – – – – – – – – 28
1.10.5 Erythropoietin (EPO) – – – – – – – 29
1.11 Phytochemicals – – – – – – – – 30
1.11.1 Phytochemical constituents in plants – – – – – 30
1.11.1.1 Alkaloids – – – – – – – – – 31
1.11.1.2 Flavonoids – – – – – – – – – 31
1.11.1.3 Tannins – – – – – – – – – 32
1.11.1.4 Steroids – – – – – – – – – 33
1.11.1.5 Terpenoids – – – – – – – – – 33
1.11.1.6 Saponins – – – – – – – – – 34
1.12 Medicinal plants – – – – – – – – 34
1.12.1 Ficus capensis – – – – – – – – 35
1.12.1.1 Botanical outline of Ficus capensis – – – – – 35
1.12.1.2 Uses of Ficus capensis – – – – – – – 35
1.12.1.3 Taxonomy of Ficus capensis – – – – – – 37
1.13 Aim and Objectives of the study – – – – – – 37
1.13.1 Aim of the study – – – – – – – – 37
1.13.2 Specific objectives of the study – – – – – – 37
CHAPTER TWO: MATERIALS AND METHODS
2.1 Materials – – – – – – – – – 38
2.1.1 Plant material – – – – – – – – 38
2.1.2 Animal – – – – – – – – 38
2.1.3 Chemicals and Reagents – – – – – – – 38
2.1.3.1 Chemicals – – – – – – – – – 38
2.1.3.2 Reagents – – – – – – – – – 39
2.2 Methods – – – – – – – – – 41
2.2.1 Aqueous extraction- – – – – – – – 41
2.2.2 Determination of extract yield – – – – – 41
2.2.3 Phytochemical analysis – – – – – – – 41
2.2.3.1 Test for Terpenoids – – – – – – – 41
2.2.3.2 Test for Glycoside – – – – – – – – 41
2.2.3.3 Test for Tanin – – – – – – – 41
2.2.3.4 Test for Cyanide – – – – – – – – 42
2.2.3.5 Test for Soluble carbohydrate – – – – – – 42
2.2.3.6 Test for Steroid – – – – – – – – 42
2.2.3.7 Test for Saponin – – – – – – – – 42
2.2.3.8 Test for Flavonoid – – – – – – – – 42
2.2.3.9 Test for Reducing sugar – – – – – – – 42
2.2.3.10 Test for Alkaloid – – – – – – – – 43
2.2.3.11 Test for Oil – – – – – – – – – 43
2.2.4 Acute toxicity and lethality – – – – – – 43
2.2.5 Induction of ulcer – – – – – – – – 43
2.2.5.1 Experimental design of ulcer – – – – – – 44
2.2.5.2 Ulcer index – – – – – – – – – 44
2.2.6 Determination of lipid peroxidation (Malondialdehyde) concentration – 45
2.2.7 Assay of catalase activity – – – – – – – 45
2.2.8 Assay of superoxide dismutase (SOD) activity – – – – 46
2.2.9 Determination of glutathione (GSH) concentration – – – 47
2.2.10 Immunomodulatory methods – – – – – 48
2.2.10.1 Determination of delayed- Type Hypersensitivity (DTH) reaction – 48
2.2.10.2 Determination of Antibody titration (Humoral mediated response) – 48
2.2.10.3 Determination of cyclophosphamide induced myelosuppression in rats – 49
2.2.10.3.1Experimental Design of cyclophosphamide induced myelosuppression in rats 49
2.2.10.3.2Determination of packed cell volume – – – – – 50
2.2.10.3.3Determination of haemoglobin (Hb) concentration – – – 50
2.2.10.3.4Determination of Red Blood Cells counts (RBCs) – – – – 51
2.2.10.3.5Determination of Total White Blood Cell count – – – – 52
2.3 Statistical Analysis – – – – – – – – 53
CHAPTER THREE: RESULTS
3.1 Percentage yield of Aqueous Leaf Extract of Ficuscapensis – 54
3.2 Phytochemical Analysis – – – – – – – 55
3.2.1 Qualitative Phytochemical Screening of Ficuscapensis – – – 55
3.2.2 Quantitative Phytochemical Constituents of Aqueous Leaf Extract
of F. capensis – – – – – – – – – 57
3.3 Biological Effects of F. capensis – – – – – – 59
3.3.1 Acute Toxicity (LD50) test of aqueous extract of the leaves of F. capensis 59
3.3.2 Ulcerogenic Activity of the Extract – – – – – 61
3.3.2.1 Ulcer Index and Preventive Ratio of aqueous extract of the leaves of Capensis – – – – – – – – – 61
3.3.3 Enzymatic antioxidant – – – – – – – 63
3.3.3.1 Effect of aqueous extract of Ficus capensis leaves on Mean Catalase Activity on Diclophenac sodium-induced Gastric Lesion in Wistar Albino Rats — – – – – – – – 63
3.3.3.2 Effect of aqueous extract of Ficus capensis leaves on Mean Malondialdehyde (MDA) concentration on Diclophenac sodium-induced Gastric Lesion in Wistar Albino Rats – – – – – – – – 65
3.3.3.3 Effect of aqueous extract of Ficus capensis leaves on Mean superoxide dismutase (SOD) Activity on Diclophenac sodium-induced Gastric Lesion in Wistar Albino Rats – – – – – – 67
3.3.3.4 Effect of aqueous extract of Ficus capensis leaves on Mean Glutathione (GSH) Activity on Diclophenac sodium-induced Gastric Lesion in Wistar Albino Rats – – – – – – – – – 69
3.4 Immunomodulatory Activities of the Extract – – – – 71
3.4.1 Effect of the extract on Delayed – Type Hypersensitivity (DTH) reaction- 71
3.4.2 Effect of the extract on Humoral antibody synthesis – – – 73
3.4.3 Cyclophosphamide Induced myelosuppression – – – – 75
3.4.3.1 Treatment with Cyclophosphamide – – – – – 75
3.4.3.2 Treatment with levamisol – – – – – – – 75
3.4.3.3 Effects of aqueous extract on cyclophosphamide induced myelosuppression on the Hb concentration of rats before and after induction with treatments 75
3.4.3.4 Effects of aqueous extract on cyclophosphamide induced myelosuppression on the RBC of rats before and after induction with treatments – – 77
3.4.3.5 Effects of aqueous extract on cyclophosphamide induced myelosuppression on the PCV of rats before and after induction with treatments – – 79
3.4.3.6 Effects of aqueous extract on cyclophosphamide induced myelosuppression on the WBC of rats before and after induction with treatments – – 81
CHAPTER FOUR: DISCUSSION
4.1 Conclusion – – – – – – – – – 92
4.2 Suggestions for Further studies – – – – – – 92
REFERENCES – – – – – – – – – 93
APPENDICES – – – – – – – – – 109
INTRODUCTION
Plant materials are sources of shelter, food and medicinal compounds which have been known to play a dominant role in the maintenance of human health in most rural communities in the developing countries (Oduola et al., 2005).
Herbal medicine is fast emerging as an alternative treatment to available synthetic drugs for the treatment of diseases possibly due to lower cost, availability, fewer adverse effects and perceived effectiveness (Ubaka et al., 2010).
The World Health Organisation (WHO) has shown great interest in plant derived medicines which have been described in the folklore medicines of many countries (Mukheerjee, 2002).
However, the historic role of medicinal plants in the treatment and prevention of diseases and their role as catalysts in the development of pharmacology do not however, assure their safety for uncontrolled use by an uninformed public (Matthews et al., 1999).
In Enugu State of Nigeria,many plants are used in herbal medicine to treat diseases, as well as to promote rapid healing of wound and sores (Dimo et al., 2002).
REFERENCES
Adebayo-Tayo, R. and Odeniyi, A. (2012). Phytochemicals screening and Microbial inhibitory Activities of Ficus capensis. African Journal of Biomedical Research, 15: 35-40.
Adeshina, G.O., Okeke C.E., Osuagwu, N.O., Ehinmidu J.O. (2010). Preliminary in-vitro antibacterial activities of ethanolic extracts of Ficus sycomorus Linn and Ficus platyphylla Del. (Moraceae) African Journal of MicrobiologyResearch, 4(8): 598-601.
Adreoli, T., Chan, P. D., Cowell, J.C., Gilbert, D. M., Green, G., Johnson, M. and Kasper, D.(2008). Management of Patients with Gastric and Duodenal Disorders. In: Brunner and 109 Suddarth; Textbook of Medical Nursing. 11th Edn. Elserview, Philaldephia, USA. Pp. 1203-1279.
Aebi, H.E. (1983). Catalase. In: Methods of Enzymatic Analysis. 3rd Edition. Academic Press, New York. Pp. 673-644.
Aguwa, C.N. and Ukwe, C. (1997). Gastrointestinal activities of Sterculia tragacantha leaf extracts. Phytotherapia, 68(2): 127-131.
CSN Team.
