Diabetes Mellitus (DM) is one of the fastest-growing global health emergencies of the 21st century, affecting more than half a billion people today.
Type 2 DM is caused by a combination of resistance to insulin action (predominant) and varying degrees of inadequate compensatory insulin secretory response or increased glucose production.
The management of NIDDM includes both pharmacological and non-pharmacological interventions, however, the use of OHAs, i.e., the former approach, is the main stay of NIDDM management.
Various anti-diabetic medicinal plants are sought as alternatives to OHAs, and their use is recommended by WHO with extracts obtained from different plant parts reported to manage NIDDM.
This study objectively evaluated the ameliorative effects of
The ameliorative effects of
Do the methanolic and total crude extracts of
Do the methanolic and total crude extracts of
This was an experimental laboratory based in-vivo preclinical study conducted at the Department of Pharmacology & Therapeutics, School of Biomedical Sciences, Makerere University College of Health Sciences located at Mulago Hospital complex, Kampala, Uganda.
Convenient selection of source of
A leaf of the avocado plant from which the fruits were obtained was examined by a taxonomist to identify the specific species by its local name, common name, scientific name and morphological descriptions at the Makerere University herbarium. A voucher specimen was prepared and a reference number (MY 001M), was given for future reference with the specimen kept in Makerere University herbarium.
The succulent fleshy parts of the avocado fruit were removed using a clean stainless-steel knife to obtain the seeds which were then sliced into smaller pieces using a knife and air-dried under a shed to a constant weight.
The dry slices of
Percentage yield = Weight of the concentrated extract obtained (g) / (Weight of the plant powder used (g)) * 100.
The weighed mass of
This was calculated using Mead’s resource equation. The minimum number of rats per group, n = (10/6) +1 ≈ 3 rats per group. The maximum number of rats per group, n = (20/6) +1 ≈ 5 rats per group. Therefore 5 rats were selected per group.
Wistar albino male rats of 8 (±1) weeks old were included into the study following assessment of their FBG level on the 24th day of the study with the cut off set at ≥ 250 mg/dL.
The sickly looking, malnourished appearing Wistar albino rats and those out of the 8 ±1 weeks and 100 to 150 g, age and weight ranges respectively were excluded from this study.
The rats were randomly assigned to six different groups using a simple random sampling technique. These rats were kept in wooden cages of ≈450 square cm floor area under standard conditions of 12 hours dark and light cycle, at a temperature of 25 ±2 0C and relative humidity of about 40-70% for 7 days to enable acclimatization.
Streptozotocin (99% potency, Med Chem Express-USA) solution for injection was freshly prepared by dissolving 232.738mg of the powder in 11.65 ml of cold normal saline solution to make a stock solution at a concentration of 20mg/ml. Each rat intraperitoneally received 60mg/kg body weight of this solution with the aid of a 2ml, 23-gauge needle following a fasting period of 6 hours. Each rat was then orally given 1ml of a distilled water mixed with glucose. Initial assessment for the development of DM following routine monitoring of the rats was done 48 hours later and confirmation of the development of DM was made after another 24 hours, on day 24 of the study. Note that day 1 of the study was considered as the day the rats were obtained before they were acclimatized. Assessment was done by drawing blood from the rat tails and taking a reading using an On call plus glucometer and On call plus glucose strips (Acon Labs Inc, USA) to assess the blood glucose concentration following a 6-hour fasting period. Only rats whose FBG levels was or exceeded 250mg/dL were considered diabetic. Dosing of these rats with the various study interventions including distilled water and glibenclamide as the negative and positive controls as well as the lower (200mg/kg body weight) and higher (400mg/kg body weight) doses of both the methanolic and total crude extracts was started on the same day (day 24 of the study) of confirmation of their diabetic status and continued for another 7 days following the assessment of their baseline biochemical and hematological parameters i.e., parameters from blood samples taken on day 24 after confirmation of DM status.
These in addition to each rat’s body weight were assessed as follows; two separate blood samples of approximately 0.5 to 1ml were collected in separate vacutainers from a pricked tail following a fasting period of 6 hours. The rats were, however, given access to drinking water during the 6 hours of fasting.
Vacutainers containing Ethylene Diamine Tetra Acetate (EDTA) were used to collect the blood samples of about 0.8 to 1ml which were then mixed by inversion and then analyzed using an automatic hematology analyzer (SYS MIX, Japan) at St Jude laboratories located in Luzira, Kampala, Uganda.
Blood samples of about 0.8 to 1ml were collected in vacutainers which were then centrifuged (Eppendorf® Minispin® model SPIN 1.000, Hamburg, Germany) for 5 minutes at a g force of 769m/s2 and then processed using a spectrophotometer (Shimadzu, Japan) at the same laboratory for the assessment of the levels of triglycerides, total cholesterol, LDL and HDL parameters.
Each rat was individually weighed using a calibrated weighing scale (KERN EW 600- 2M, Kern & Sohn, Germany). The healthy rats were then continued on the pellet diet, but the daily meal was doubled to increase calorie intake for two weeks.
The study interventions were orally administered using an oral intragastric tube with the dose per unit body weight. The diabetic control group, each rat received 1ml of distilled water per day for the entire 7 days.
For the ME200 group, ≈716mg of the ME was dissolved in 52ml of distilled water to make a concentration of ≈14mg/ml and each rat received a given volume of the mixture based on its baseline body weight per day for the 7 days.
For the ME400 group, ≈1,567mg of the ME was dissolved in 113ml of distilled water to make a concentration of ≈14mg/ml and each rat received a given volume of the mixture based on its baseline body weight per day for the 7 days.
For the TCE200 group, ≈706mg of the TCE was dissolved in 66ml of distilled water to make a concentration of ≈11mg/ml and each rat received a given volume of the mixture based on its baseline body weight per day for the 7 days.
For TCE400 group, ≈1611mg of the TCE was dissolved in 151ml of distilled water to make a concentration of ≈ 11mg/ml and each rat received a given volume of the mixture based on its baseline body weight per day for the 7 days.
For the GL10 group, 8 tablets of glibenclamide, 5mg strength were finely crashed and dissolved in 32ml of distilled water to make a concentration of ≈1.25mg/ml. Each rat received a given volume of the mixture based on its baseline body weight per day for the 7 days.
Following the 7-day treatment period i.e., on day 31 of the study, the biochemical and hematological parameters as well as body weight were reassessed. The rats were then euthanized on the same day by intraperitoneally injecting each rat with a combination of Ketamine and xylazine at doses of 300 mg and 30 mg/kg body weight respectively followed by cervical dislocation. The doses of the euthanizing agents were calculated based on the body weights measured on day 31 of the study.
The dependent variables included the biochemical and hematological parameters measured from each rat in each group whereas independent variables included the interventions that were orally administered to the study animals.
Data was entered excel and the mean, standard deviation as well as the standard error of the mean were calculated. Changes during the study course, including the 3 different sampling intervals i.e., Days, 7, 24 and 31, for each group were independently compared using paired (dependent) T-tests. Multiple group comparisons were performed for the extract groups alone (4 group comparisons) as well as with glibenclamide (5 group comparisons) using Welch’s ANOVA and finally, comparisons between the various groups considering the change in the mean levels of the selected parameters between day 24 and day 31 were done using independent T-test, assuming unequal variances. Statistical significance was considered at a p-value of 0.05.
This work was presented to the Department of Pharmacology and Therapeutics which approved it for ethical consideration under the school of biomedical Sciences research and ethics committees. The proposed protocol was also submitted to the Institutional Animal Care and Use Committee of the College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB) located at Makerere University Kampala, Uganda for approval prior to the conduct of the study.
The mean FBG levels in the negative diabetic control (DC) group exhibited persistent significant elevation throughout the study. Both doses of the two extract types like glibenclamide significantly improved the FBG by lowering it from the elevated levels recorded on day 24, following induction with streptozotocin (STZ). The improvement in FBG levels by both doses of the ME and TCE200 was significantly less than that of glibenclamide, however, the higher doses of both extracts were significantly better than the lower doses (
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133.44 ± 3.76 |
295.15 ± 5.87 |
322.84 ± 2.84 (9.4%↑) |
*0.00 |
*0.00 |
*0.04 |
|
|
129.76 ± 1.31 |
298.36 ± 6.05 |
141.65 ± 2.19 (52.5%↓) |
*0.00 |
*0.04 |
*0.00 |
|
|
126.07 ± 1.22 |
317.07 ± 5.20 |
133.26 ± 1.65 (60%↓) |
*0.00 |
0.09 |
*0.00 |
|
|
130.57 ± 1.43 |
287.23 ± 6.02 |
128.65 ± 1.64 (55.2%↓) |
*0.00 |
1.00 |
*0.00 |
|
|
131.96 ± 2.43 |
302.43 ± 6.12 |
114.69 ± 4.72 (62.1%↓) |
*0.00 |
*0.01 |
*0.00 |
|
|
128.73 ± 0.76 |
310.11 ± 7.06 |
109.37 ± 2.98 (64.7%↓) |
*0.00 |
*0.01 |
*0.00 |
DC: Diabetic Control, ME: Methanolic Extract, TCE: Total Crude Extract, GL10: Glibenclamide [Values are expressed as mean ± SEM (n=5), Percentage increase (↑) or decrease (↓), *significant at p<0.05]
The mean HDL inversely correlated with LDL, TC and TG, with the mean TC, TG and LDL levels in the negative DC group exhibiting persistent significant increase throughout the study. All extracts significantly improved the TC levels when compared to glibenclamide but ME400 was significantly better than ME200.
The effects of glibenclamide on the mean TG levels unlike that of the extracts was not significant.
Glibenclamide, like the extracts caused significant improvements in the LDL levels, however, the effects of the TCEs were significantly better than that of glibenclamide.
The improvement in HDL levels by all extracts unlike glibenclamide was significant but still comparable to that of the latter as well as to each other (
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68.43 ± 4.04 |
131.88 ± 1.30 |
159.43 ± 1.28 (20.9%↑) |
*0.00 |
*0.00 |
*0.00 |
|
|
65.65 ± 0.58 |
125.24 ± 1.91 |
98.14 ± 1.80 (21.6%↓) |
*0.001 |
*0.001 |
*0.02 |
|
|
69.66 ± 1.83 |
128.56 ± 1.52 |
82.42 ± 0.70 (35.9%↓) |
*0.00 |
*0.001 |
*0.00 |
|
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69.37 ± 3.98d |
130.11 ± 1.87 |
78.80 ± 1.11 (39.4%↓) |
0.001 |
0.18 |
*0.00 |
|
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70.22 ± 2.34 |
127.48 ± 1.17 |
74.06 ± 0.92 (41.9%↓) |
*0.00 |
0.48 |
*0.00 |
|
|
65.28 ± 1.24 |
125.13 ± 2.22 |
101.84 ± 3.09 (18.6%↓) |
*0.00 |
*0.003 |
0.09 |
|
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|
|||||
|
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84.11 ± 1.17 |
108.45 ± 2.95 |
123.88 ± 4.97 (14.2%↑) |
*0.003 |
*0.003 |
0.06 |
|
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83.67 ± 0.76 |
105.04 ± 2.37 |
88.22 ± 0.66 (16.0%↓) |
*0.003 |
0.15 |
*0.01 |
|
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87.01 ± 1.35 |
110.81 ± 2.90 |
85.62 ± 1.80 (22.7%↓) |
*0.01 |
0.81 |
**0.001 |
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85.75 ± 1.01 |
114.05 ± 4.07 |
86.82 ± 1.25 (23.9%↓) |
*0.003 |
0.66 |
*0.003 |
|
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84.32 ± 1.37 |
100.65 ± 0.46 |
83.09 ± 1.06 (17.5%↓) |
*0.003 |
1.00 |
*0.00 |
|
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86.54 ± 1.60 |
109.24 ± 3.19 |
93.18 ± 2.30 (14.7%↓) |
*0.03 |
*0.03 |
0.09 |
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37.12 ± 1.08 |
94.60 ± 1.23 |
106.88 ± 3.28 (13.0%↑) |
*0.00 |
*0.00 |
*0.01 |
|
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38.54 ± 1.74 |
87.14 ± 1.94 |
45.82 ± 1.76 (47.4%↓) |
*0.00 |
0.06 |
*0.00 |
|
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35.34 ± 1.22 |
88.33 ± 1.11 |
33.62 ± 1.62 (61.9%↓) |
*0.00 |
0.24 |
*0.00 |
|
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38.96 ± 1.79 |
90.22 ± 2.63 |
36.78 ± 1.03 (59.2%↓) |
*0.00 |
0.96 |
*0.00 |
|
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36.22 ± 0.51 |
93.48 ± 2.79 |
33.06 ± 1.06 (64.6%↓) |
*0.00 |
0.06 |
*0.00 |
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36.38 ± 1.27 |
95.27 ± 4.12 |
58.98 ± 1.81 (38.1%↓) |
*0.00 |
*0.001 |
*0.001 |
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36.51 ± 0.65 |
30.61 ± 1.34 |
29.88 ± 1.05 (2.4%↓) |
0.09 |
*0.02 |
0.66 |
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37.50 ± 1.78 |
30.56 ± 0.40 |
35.99 ± 1.00 (17.8%↑) |
*0.003 |
1.00 |
*0.03 |
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37.88 ± 1.23 |
32.40 ± 0.70 |
39.62 ± 1.04 (22.3%↑) |
0.06 |
1.00 |
*0.02 |
|
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36.96 ± 0.69 |
29.98 ± 0.87 |
36.81 ± 1.03 (22.8%↑) |
*0.001 |
1.00 |
*0.03 |
|
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34.27 ± 1.16 |
31.11 ± 0.38 |
38.04 ± 1.17 (22.3%↑) |
0.12 |
0.18 |
*0.003 |
|
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35.37 ± 0.55 |
31.75 ± 0.40 |
34.22 ± 1.31 (14.1%↑) |
*0.003 |
*0.04 |
0.06 |
DC: Diabetic Control, ME: Methanolic Extract, TCE: Total Crude Extract, GL10: Glibenclamide [Values are expressed as mean ± SEM (n=5), Percentage increase (↑) or decrease (↓), *significant at p<0.05].
The correlations among the various hematological parameters were significant with the strongest being between MCV and Hct (r=0.99) and the weakest being between RBCs and MCHC (r=0.76).
The mean RBC, MCH, MCHC, MCV, Hbg and Hct levels in the negative DC group on day 31 were significantly (p=0.00) lower than the levels recorded on day 7 with the TCEs exhibiting a remarkable improvement in the mean RBC counts. The effect of all extracts on MCH levels were significantly better than that of glibenclamide but only the higher doses of both extract types caused a significant improvement in the mean MCHC (p=0.001 and p=0.01) for ME and TCE respectively. All extracts significantly improved the mean MCV levels with ME400 and the TCEs having a significantly better activity than glibenclamide. All extracts unlike glibenclamide significantly improved the mean Hbg and Hct levels and their effect was significantly better than that of glibenclamide (
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7.13 ± 0.14 |
5.90 ± 0.23 |
5.00 ± 0.16 (15.3%↓) |
*0.01 |
*0.00 |
0.06 |
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6.92 ± 0.24 |
5.43 ± 0.05 |
6.12 ± 0.31 (12.7%↑) |
*0.01 |
0.21 |
0.21 |
|
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7.05 ± 0.21 |
6.04 ± 0.18 |
6.93 ± 0.48 (14.7%↑) |
0.14 |
1.00 |
0.42 |
|
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7.16 ± 0.52 |
5.75 ± 0.20 |
7.01 ± 0.60 (21.9%↑) |
0.15 |
1.00 |
0.24 |
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7.02 ± 0.25 |
5.22 ± 0.29 |
7.40 ± 0.67 (41.8%↑) |
0.06 |
1.00 |
*0.03 |
|
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7.21 ± 0.21 |
5.48 ± 0.17 |
6.00 ± 0.18 (10.5%↑) |
*0.001 |
*0.001 |
0.21 |
|
MCH (mean |
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19.34 ± 0.46 |
16.2 ± 0.52 |
11.09 ± 0.93 (31.5%↓) |
*0.03 |
*0.001 |
*0.03 |
|
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19.03 ± 0.48 |
16.92 ± 0.84 |
18.82 ± 0.75 (11.2%↑) |
0.12 |
1.00 |
0.42 |
|
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18.44 ± 0.68 |
17.33 ± 0.81 |
19.61 ± 0.30 (13.2%↑) |
1.00 |
0.21 |
0.12 |
|
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19.15 ± 0.41 |
16.49 ± 0.82 |
17.80 ± 0.66 (7.9%↑) |
0.24 |
1.00 |
0.30 |
|
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18.50 ± 0.47 |
17.08 ± 0.59 |
19.66 ± 1.61 (15.1%↑) |
0.45 |
1.00 |
0.33 |
|
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17.89 ± 0.64 |
18.10 ± 0.87 |
16.02 ± 0.48 (11.5%↓) |
1.00 |
0.48 |
0.15 |
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||||||
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30.14 ± 0.33 |
26.34 ± 0.95 |
19.46 ± 1.77 (26.1%↓) |
0.06 |
*0.01 |
0.06 |
|
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31.32 ± 0.56 |
27.56 ± 0.93 |
30.54 ± 0.58 (10.8%↑) |
0.06 |
0.45 |
0.27 |
|
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30.74± 0.47 |
27.21 ± 0.711 |
32.60 ± 1.04 (19.8%↑) |
*0.03 |
0.45 |
*0.001 |
|
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31.22 ± 0.32 |
25.38 ± 0.67 |
30.66 ± 0.21 (21.0%↑) |
*0.01 |
1.00 |
0.09 |
|
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30.98 ± 0.35 |
26.85 ± 1.12 |
33.40 ± 1.73 (24.3%↑) |
0.06 |
1.00 |
*0.01 |
|
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31.11 ± 0.44 |
27.44 ± 0.85 |
30.62 ± 2.14 (11.6%↑) |
*0.03 |
1.00 |
0.6 |
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71.21 ± 0.68 |
62.17 ± 1.11 |
47.26 ± 0.82 (24%↓) |
*0.01 |
*0.00 |
*0.003 |
|
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72.43 ± 0.70 |
60.33 ± 0.77 |
67.14 ± 1.31 (11.3%↑) |
*0.01 |
*0.03 |
*0.03 |
|
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70.60 ± 1.06 |
59.87 ± 1.40 |
71.27 ± 1.87 (19.0%↑) |
*0.003 |
1.00 |
*0.03 |
|
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73.29 ± 1.06 |
61.05 ± 0.39 |
71.46 ± 1.31 (17.1%↑) |
*0.002 |
0.03 |
*0.01 |
|
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72.61 ± 0.47 |
60.79 ± 0.59 |
73.22 ± 0.86 (20.5%↑) |
*0.001 |
1.00 |
*0.003 |
|
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72.18 ± 0.40 |
60.00 ± 0.62 |
63.62 ± 0.96 (6.0%↑) |
*0.00 |
0.002 |
0.21 |
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||||||
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15.33 ± 0.29 |
12.06 ± 0.54 |
9.20 ± 0.25 (23.7%↓) |
0.05 |
*0.00 |
*0.03 |
|
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15.24 ± 0.53 |
11.21 ± 0.38 |
15.76 ± 1.08 (40.6%↑) |
*0.03 |
1.00 |
*0.02 |
|
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17.01 ± 0.86 |
11.64 ± 0.43 |
16.91 ± 0.92 (45.3%↑) |
*0.003 |
1.00 |
*0.01 |
|
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15.43 ± 0.34 |
12.36 ± 0.53 |
16.02 ± 0.69 (29.6%↑) |
*0.01 |
1.00 |
*0.02 |
|
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16.03 ± 0.49 |
11.68 ± 0.35 |
17.04 ± 0.51 (45.9%↑) |
*0.03 |
1.00 |
*0.00 |
|
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16.50 ± 0.22 |
11.37 ± 0.46 |
15.05 ± 0.34 (32.4%↑) |
*0.001 |
0.16 |
0.09 |
|
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||||||
|
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45.22 ± 0.99 |
34.50 ± 0.48 |
27.66 ± 1.10 (19.8%↓) |
*0.003 |
*0.001 |
*0.01 |
|
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44.38 ± 0.56 |
33.81 ± 0.83 |
42.14 ± 1.49 (24.6%↑) |
*0.003 |
0.48 |
*0.03 |
|
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45.14 ± 0.94 |
33.78 ± 1.02 |
45.63 ± 1.68 (35.0%↑) |
*0.003 |
1.00 |
*0.003 |
|
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45.57 ± 0.78 |
34.54 ± 0.83 |
43.46 ± 1.14 (25.8%↑) |
*0.003 |
0.63 |
*0.001 |
|
|
46.01 ± 0.71 |
34.33 ± 1.19 |
46.20 ± 1.60 (34.6%↑) |
*0.01 |
1.00 |
*0.01 |
|
|
45.27 ± 1.04 |
36.58 ± 1.45 |
40.22 ± 2.80 (10.0%↑) |
*0.01 |
0.20 |
0.84 |
DC: Diabetic Control, ME: Methanolic Extract, TCE: Total Crude Extract, GL10: Glibenclamide [Values are expressed as mean ± SEM (n=5), Percentage increase (↑) or decrease (↓), *significant at p<0.05].
The effects on FBG levels for comparisons among the extract treated groups alone and with glibenclamide on day 31 were significant with p-values, 0.001 and 0.00* respectively. TCE400 exhibited the greatest and comparable effects to glibenclamide (p=0.31) and ME400 (p=0.08) but significantly better activity than TCE200 (p=0.01) and ME200 (p=0.003). The FBG effect for TCE200 was comparable to both doses of the ME200 (p=0.09) and ME400 (p=0.08).
Comparisons in the mean TG levels among the extract treated groups were significant (p=0.00). TCE400 exhibited the greatest but comparable effects to TCE200 (p=0.08) but a significantly greater activity than ME400 (p=0.001) and ME200 (p=0.00). TCE200 exerted a significantly better effect than both doses of the ME. Group comparisons regarding the mean TC levels were significant (p=0.03). TCE200 exhibited the greatest but comparable effect to ME400 (p=0.71) as well as a significantly greater activity than TCE400 (p=0.04) and ME200 (p=0.03). ME400 exerted a significantly better effect than TCE400 (p=0.02).
Group comparisons regarding the mean LDL levels were significant (p=0.003). TCE (400) exhibited the greatest effect which was comparable to ME400 (p=0.37) and TCE200 (p=0.05) but significantly greater than ME200 (p=0.001). TCE 200 exerted a significantly better effect than ME200 (p=0.01), however, its activity was comparable to that of ME400 (p=0.36).
Extract group comparisons for the mean RBC levels were significant (p=0.04) but only TCE400 showed significant (p=0.03) improvement in the RBC count with its activity comparable to TCE200 (p=0.13) but a significantly better than that of ME400 (p=0.03) and ME200 (p=0.01). The effect of TCE200 was comparable to that of the MEs.
Comparisons for the mean MCH levels among the extract treated groups were significant (p=0.03). TCE400 exhibited a comparable effect to both ME400 (p=0.95) and ME200 (p=0.83). TCE200 also showed a comparable activity to both doses of the MEs. Only ME400 and TCE400 demonstrated significant improvement in the mean MCHC levels with (p=0.001 and 0.01) respectively. TCE200 showed a significantly better activity than both doses of the ME200 (p=0.02) and ME400 (p=0.04). Extract group comparisons regarding the mean MCV levels were significant (p=0.03). The effect of TCE400 was significantly (p=0.03) greater than that of ME200, however, the effects of all extracts on the mean Hbg and Hct levels were comparable i.e., (0.08 ≤ p ≤0.92) and (0.12 ≤ p ≤ 0.92) respectively.
The yield of
The total crude extract obtained a comparatively higher yield than the methanolic and this may be attributed to its wider extraction potential for both polar and non-polar components of the plant material.
The chemical used, STZ induced DM through the destruction of the pancreatic islet β-cell which reduced the population of functional beta cells eventually resulting in hyperglycemia and hence DM.
Both extracts of
Higher doses of both extract types performed better than the lower ones, a concept of dose dependent potency observed in other studies.
Streptozotocin induces changes in lipid profiles as early as after 24 hours just as was observed in this study.
The TCEs and ME400 in most instances exhibited significantly better lipid profile improvements than glibenclamide, findings consistent with another study.
The extracts of
Almost all the various hematological indices showed a significant decrease after STZ administration possibly due to the increased destruction of RBC membranes with subsequent rupture of the cells under the influence of NIDDM induced oxidative stress.
The administration of the
Findings from our study are like those of a study by Obiandu et al (2022) where some hematological indices, specifically RBC count, Hbg and Hct were significantly improved following the administration of
The two different doses of both extracts exhibited positive changes on both the biochemical and hematological parameters in the diabetic rats at variable magnitudes with the higher doses generally performing better. The ability of the extracts to improve the STZ induced derangements in these selected parameters to levels comparable to those of the healthy non-diabetic rats (baseline parameters taken on day 7) highlighted their ameliorative capability.
Appropriately selected doses of the seed extracts of
My sincere appreciation is expressed to my supervisors Dr. Norah Mwebaza and Dr. Isaac Magulu Kimbowa of the Pharmacology and Therapeutics Department for their support, guidance and sacrifice of their time amidst their busy schedules. Their shared wisdom and insight concerning life and education is greatly appreciated. I express my gratitude and appreciation to my family for their relentless encouragement and above all prayers for me to excel in my studies. I would like to thank the almighty God above all for giving me the strength and grace to write this paper.