- The document analyzes the effects of aqueous extracts of Picralima nitida and Rauvolfia vomitoria, alone and in combination, on blood glucose levels and liver enzyme activity in alloxan-induced diabetic rats. - The extracts were found to contain alkaloids, cardenolides, and saponins. Administration of the extracts significantly reduced blood glucose and liver enzyme levels compared to the diabetic control group. - The combination of P. nitida and R. vomitoria extracts had a comparable hypoglycemic effect to P. nitida alone and more effectively reduced liver enzyme activity, suggesting it could be an effective treatment for diabetes and its complications.

1. IMMANUEL OLUWAFEMI .T
2007/0875
DEPARTMENT OF BIOCHEMISTRY
COLLEGE OF NATURAL SCIENCES
UNIVERSITY OF AGRICULTURE ABEOKUTA, OGUN STATE.
JUNE, 2011

2. • Diabetes mellitus is a group of metabolic diseases characterized
by high blood sugar (glucose) level that result from defects in
insulin secretion or action.
• An ideal oral treatment for diabetes would be a drug
(glibenclamide) which controls glycemic level and development
of complications.
• Alloxan induces diabetes in experimental animals through beta
cells destruction (Singh and Gupta, 2007).
• (AST & ALT) aminotransaminases are liver enzymes, their high
activity in the plasma indicate the damage of the liver.
• Herbal medicine is the oldest form of healthcare known to
mankind and herbs had been used to cure all form of diseases or
ailments (Rawat, 2003).
• Picralima nitida is a monotypic plant exploited for it mood and
equally employed as an arrow in fish poison (Omino, 2002). It
belongs to the family apocynaceae in East Africa and studies have
shown that it have some opioid analgesics activities (Menzies et
al., 1998), some hypoglycemic effects (Inya-Agha, 1999), and
antimicrobial properties (Fakeye et al., 2004).

3. • Rauvolfia vomitoria has been investigated for alkaloid
content, especially for those with hypotensive and anti-
inflammatory properties (Chatterjee and Bandyopadhyay,
1979; Amer and Court, 1980; Kweifio-Okai, 1991).
AIMS OF RESEARCH
The objectives of this work include:
• To determine the presence of alkaloids, cardenolides,
anthraquinones, saponins, tannins, and phenols
• To determine the biochemical effect of aqueous extract of
picralima nitida and rauvolfia vomitoria on plasma and liver
(AST &ALT) aminotransferase of an alloxan induced male
albino rats.

4. • Fifty-five male albino rats were bought and feed with food and
water ad libitum for four weeks. Forty of the rats were induced
with diabetes with alloxan monohydrate. The rats were grouped
into eleven groups, which include:
• Grp 1 No diabetes, No treatment
• Grp 2 No diabetes + 300mg/kgPn
• Grp 3 No diabetes 300kgPn+Rv
• Grp 4 Diabetes, No treatment
• Grp 5 Diabetes + 150mg/kgStandard drug
• Grp 6 Diabetes +100mg/kgPn
• Grp 7 Diabetes +200mg/kgPn
• Grp 8 Diabetes +300mg/kgPn
• Grp 9 Diabetes +100mg/kgPn+Rv
• Grp 10 Diabetes +200mg/kgPn+Rv
• Grp 11 Diabetes +300mg/kgPn+Rv

5. • The seeds and roots of Picralima nitida and Rauvolfia
vomitoria were extracted using an aqueous solution after
been milled into a fine form.
• The extracts were administered orally at different
concentration for 7 days and their blood glucose were
monitored.
• The animals were sacrificed after the 8th
day and whole blood
was collected through cardiac puncture into an heparinized
tube, which was centrifuge at 4000rpm for 10minutes to
obtain plasma.
• The organs were harvested and washed in 0.9g normal saline
solution, the liver was homogenized in 0.25M of sucrose
solution and the homogenates and plasma (AST & ALT)
were assayed for using commercial assay test kit.

6. Table 1: Result of phytochemical screening of Picralima nitida and
Rauvolfia vomitoria
Test Results
Picralima nitida
Alkaloids test +ve
Cardennolides test +ve
Saponins +ve
Rauvolfia vomitoria
Alkaloids test +ve
Cardenolides test +ve
Saponins +ve

7. DAY 0 DAY 1 DAY3 DAY 8
Cn Nodiab, no
treat
105.00±21.01ab
83.40±9.37ab
77.60±14.46a
77.20±6.79abc
Cn No diab+Pn 86.75±9.03ab
88.25±10.90ab
73.50±4.43a
66.50±5.80ab
Cn Diab,notreat 67.40±9.52a
368.20±195.66cd
337.80±26.86cde
300.00±43.58d
Diab +Gibb 72.80±5.21ab
277.60±206.86bcd
313.50±37.74bcd
157.24±136.06bc
Diab+100mgPn 80.00±3.00ab
61.00±8.00a
464.00±74.00ef
88.67±40.50abc
Diab+200mgPn 68.33±13.20a
447.00±103.00d
504.33±105.15f
166.67±40.50abc
Diab+300mgPn 68.67±17.03a
376.33±167.15cd
398.00±89.26def
94.75±55.30abc
Table 2: Effect of Picralima nitida extracts on blood glucose.
Values expressed as MEAN± SD. Values within the same column
with different superscripts are significantly different at p 0.05.˂

8. GROUPS PLASMA ALT (U/l)
Cn.No Diab. No treat
Cn. Pn
Cn. Diab.No treat
Cn. Diab+glib
Ts. 100mgPn
Ts. 200mgPn
Ts. 300mgPn
2.91±0.51a
8.00±0.916b
7.40±1.25b
15.73±3.50e
31.00±1.00f
13.15±0.85de
9.26±1.75bc
TABLE 4: EFFECT OF AQUEOUS EXTRACT OF Picralima nitida ON ALT
PLASMA IN ALBINO RATS
Values expressed as MEAN± SD. Values within the same column with different
superscripts are significantly different at p 0.05.˂

9. GROUPS PLASMA ALT (U/l)
Cn.No Diab. No treat
Cn. Pn+Rv
Cn. Diab.No treat
Cn. Diab+glib
Ts. 100mgPn+Rv
Ts. 200mgPn+Rv
Ts. 300mgPn+Rv
2.91±0.51a
11.35±1.30cd
7.40±1.24b
15.73±3.50e
14.36±1.50de
16.03±1.81e
13.10±2.60de
TABLE 5: EFFECT OF COMBINED THERAPHY OF AQUEOUS EXTRACT OF
Picralima nitida AND Rauvolfia vomitoria ON ALT PLASMA IN ALBINO RATS
Values expressed as MEAN± SD. Values within the same column with different
superscripts are significantly different at p 0.05.˂

10. GROUPS LIVER ALT (U/l)
Cn.No Diab. No treat
Cn. Pn
Cn. Diab.No treat
Cn. Diab+glib
Ts. 100mgPn
Ts. 200mgPn
Ts. 300mgPn
5.43±0.63ab
7.46±1.35b
35.03±1.04e
6.40±1.10b
6.80±1.00b
53.80±1.00f
12.80±1.00cd
TABLE 6: EFFECT OF AQUEOUS EXTRACT OF Picralima nitida ON ALT
LIVER IN ALBINO RATS
Values expressed as MEAN± SD. Values within the same column with different
superscripts are significantly different at p 0.05˂

11. GROUPS LIVER ALT (U/l)
Cn.No Diab. No treat
Cn. Pn+Rv
Cn. Diab.No treat
Cn. Diab+glib
Ts. 100mgPn+Rv
Ts. 200mgPn+Rv
Ts. 300mgPn+Rv
5.43±0.63ab
14.27±2.14d
35.03±1.04e
6.40±1.10b
4.26±1.20a
4.30±0.30a
11.27±0.60c
TABLE 7: EFFECT OF COMBINED THERAPHY OF AQUEOUS EXTRACT
OF Picralima nitida AND Rauvolfia vomitoria ON ALT LIVER IN ALBINO
RATS
Values expressed as MEAN± SD. Values within the same column with different
superscripts are significantly different at p 0.05˂

12. GROUPS PLASMA AST (U/l)
Cn.No Diab. No treat
Cn. Pn
Cn. Diab.No treat
Cn. Diab+glib
Ts. 100mgPn
Ts. 200mgPn
Ts. 300mgPn
45.20±11.83a
40.06±7.04a
27.56±11.50a
119.03±14.77b
137.33±21.35b
66.50±1.00a
128.70±43.69b
TABLE 8: EFFECT OF AQUEOUS EXTRACT OF Picralima nitida ON AST
PLASMA IN ALBINO RATS
Values expressed as MEAN± SD. Values within the same column with
different superscripts are significantly different at p 0.05˂

13. GROUPS PLASMA AST (U/l)
Cn.No Diab. No treat
Cn. Pn+Rv
Cn. Diab.No treat
Cn. Diab+glib
Ts. 100mgPn+Rv
Ts. 200mgPn+Rv
Ts. 300mgPn+Rv
45.20±11.83a
25.03±5.21a
27.56±11.50a
119.03±14.77b
238.77±29.15f
115.13±32.02b
189.20±42.70c
TABLE 9: EFFECT OF COMBINED THERAPHY OF AQUEOUS EXTRACT
OF Picralima nitida AND Rauvolfia vomitoria ON AST PLASMA IN ALBINO
RATS
Values expressed as MEAN± SD. Values within the same column with
different superscripts are significantly different at p 0.05˂

14. GROUPS LIVER AST (U/l)
Cn.No
DiabNotreat
Cn. Pn
Cn. Diab.No treat
Cn. Diab+glib
Ts. 100mgPn
Ts. 200mgPn
Ts. 300mgPn
62.00±3.01bc
48.00±1.00b
107.30±1.00de
12.20±2.30a
166.80±1.00f
355.80±1.00g
122.00±1.00e
TABLE 10: EFFECT OF AQUEOUS EXTRACT OF Picralima nitida ON AST
LIVER IN ALBINO RATS
Values expressed as MEAN± SD. Values within the same column with different
superscripts are significantly different at p 0.05.˂

15. GROUPS LIVER AST (U/l)
Cn.No Diab. No treat
Cn. Pn+Rv
Cn. Diab.No treat
Cn. Diab+glib
Ts. 100mgPn+Rv
Ts. 200mgPn+Rv
Ts. 300mgPn+Rv
62.00±3.01bc
161.17±57.75f
107.30±1.00de
12.20±2.30a
48.16±14.18b
87.50±1.00cd
50.30±20.05b
TABLE 11: EFFECT OF COMBINED THERAPHY OF AQUEOUS EXTRACT
OF Picralima nitida AND Rauvolfia vomitoria ON AST LIVER IN ALBINO RATS
Values expressed as MEAN± SD. Values within the same column with different
superscripts are significantly different at p 0.05.˂

16. GROUPS DAY 0 DAY 1 DAY3 DAY 8
No diab,no treat 105.00±21.01ab
83.40±9.37ab
77.60±14.46a
77.20±6.79abc
No diab Pn+Rv 76.40±14.25ab
75.40±8.20ab
69.9±8.35a
66.00±7.17ab
Diab +no treat 67.40±9.52a
368.20±195.66cd
337.80±26.86cde
300.00±43.58d
Diab +gilbb 72.80±5.21ab
277.60±206.86bcd
313.50±37.74bcd
157.24±136.06bc
Diab+100mgPn+Rv 110.00±62.83b
300.00±2.00cd
310.25±181.59bcd
58.00±2.44a
Diab+200mgPn+Rv 68.50±12.97a
234.75±166.74abc
214.00±138.268cd
75.00±27.71abc
Diab+300mgPn+Rv 78.33±13.20ab
258.67±30.50abcd
190.67±69.51ab
70.33±2.51abc
TABLE 3: EFFECT OF MIXTURE OF Picralima nitida AND Rauvolfia
vomitoria ON THE BLOOD
Values expressed as MEAN± SD. Values within the same column with
different superscripts are significantly different at p 0.05˂

17. • In this study, the phytochemical screening of Picralima nitida and Rauvolfia
vomitoria indicates the presence of alkaloids, cardenolides, and saponins. The
herbs containing some secondary metabolites have become inevitable due to
significant correlation between their traditional medical and their extractives
(Fatope, 1995)
• In this research, hypoglycemic effect of P. nitida on alloxan induced diabetic
rats was confirmed as it reduced the hyperglycemic effect of diabetic rats
(inya et al., 2006)
• The mixture of the extracts of P. nitida and R. vomitoria have a significant
hypoglycemic effect on alloxan-induced diabetes comparable to that of the P.
nitida, and could serve as an effective adjunct in the management of diabetes
mellitus.
• The mixture of the extracts reduced the increased activity of AST and ALT in
the liver and plasma compared to the rats given Picralima nitida extracts
which also reduced (AST &ALT activity) but not significant to the control
group), which was markedly elevated as a result of exposure to drugs or
plants extracts that are toxic to the liver (Bass, 1996)

18. • Bass NM, Ockner BA. (1996). Drug-induced liver disease, Hepatology: a
textbook of liver disease, 3rd
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International Journal of Pharmacology 2 (5) 576-580.
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alkaloids extracted from Picralima nitida (fam. Apocynaceae). European Journal
of Pharmacology. ;350(1):101-8.
• Paul, T., Giboney, M.D. Mildly Elevated Liver Transaminase levels in
Asymptomatic Patient, (2007). American Family Physician.
• Singh N, Gupta M (2007). Effects of ethanolic extract of Syzygium cumini
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19. • Bass NM, Ockner BA. Drug-induced liver disease, Hepatology: a
textbook of liver disease, 3rd
eds. Philadelphia: WB Saunders, 1996; 962-
1017
• Fatope, M.O. (1995) . Phytocompounds: their Bioassay and Diversity.
Discov. Inov. 7(3):229-236.
• Menzies JRW, Paterson SJ, Duwiejua M, Corbett AD. Opioid activity of
alkaloids extracted from Picralima nitida (fam. Apocynaceae). European
Journal of Pharmacology. 1998 May 29;350(1):101-8.
 N. A. Trivedi, B. Mazumdar, J. D. Bhatt, K. G. Hemavathi .(2004).
Effect of shilajit on blood glucose and lipid profile in alloxan-
induced diabetic rats. Department of Pharmacology,
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Asymptomatic Patient, (2007). American Family Physician.
•

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