IMMANUEL OLUWAFEMI .T2007/0875

DEPARTMENT OF BIOCHEMISTRYCOLLEGE OF NATURAL SCIENCES

UNIVERSITY OF AGRICULTURE ABEOKUTA, OGUN STATE.

JUNE, 2011

• 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).

• 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.

• 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

• 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.

Table 1: Result of phytochemical screening of Picralima nitida and Rauvolfia vomitoria

Test ResultsPicralima nitidaAlkaloids test +veCardennolides test +veSaponins +ve

Rauvolfia vomitoriaAlkaloids test +ve Cardenolides test +veSaponins +ve

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.

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.

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. 

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

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

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

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

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.

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.

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

• 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)

• Bass NM, Ockner BA. (1996). Drug-induced liver disease, Hepatology: a textbook of liver disease, 3rd eds. Philadelphia: WB Saunders; 962-1017

• Fakeye TO, Itiola OA, George AO, Odetola HA (2004). Antimicrobial Property of Picralima nitida Stem Bark Extract in Cream Formulations. Pharmacol. Biol. 42(4-5): 274-279.

• Fatope, M.O. (1995) . Phytocompounds: their Bioassay and Diversity. Discov. Inov. 7(3):229-236.

• Inya-Agha SI., Ezea SC., and Odukoya OA. (2006). Evaluation of Picralima nitida: Hypoglycemic activity, Toxicology and Analytical Standards. International Journal of Pharmacology 2 (5) 576-580.

• Inya-Agha SI (1999). The Hypoglycemic Properties of Picralima nitida. Niger. J. Nat. Prod. Med. 3: 66-67. Katsumat

• Menzies JRN, Paterson SJ, Duwiejua M, Corbett AD. (1998 ). Opioid activity of 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 (Linn) seed powder on pancreatic islets of alloxan diabetic rats, Indian J Exp Biol. 45(10): 861-867

• 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, MedicalCollege, Baroda -390001, India.

• Paul, T., Giboney, M.D. Mildly Elevated Liver Transaminase levels in Asymptomatic Patient, (2007). American Family Physician.

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