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Assessment of genetic diversity in Trigonella foenum-graecum and Trigonella caerulea using ISSR and RAPD markers.

BACKGROUND: Various species of genus Trigonella are important from medical and culinary aspect. Among these, Trigonella foenum-graecum is commonly grown as a vegetable. This anti-diabetic herb can lower blood glucose and cholesterol levels. Another species, Trigonella caerulea is used as food in the form of young seedlings. This herb is also used in cheese making. However, little is known about the genetic variation present in these species. In this report we describe the use of ISSR and RAPD markers to study genetic diversity in both, Trigonella foenum-graecum and Trigonella caerulea. RESULTS: Seventeen accessions of Trigonella foenum-graecum and nine accessions of Trigonella caerulea representing various countries were analyzed using ISSR and RAPD markers. Genetic diversity parameters (average number of alleles per polymorphic locus, percent polymorphism, average heterozygosity and marker index) were calculated for ISSR, RAPD and ISSR+RAPD approaches in both the species. Dendrograms were constructed using UPGMA algorithm based on the similarity index values for both Trigonella foenum-graecum and Trigonella caerulea. The UPGMA analysis showed that plants from different geographical regions were distributed in different groups in both the species. In Trigonella foenum-graecum accessions from Pakistan and Afghanistan were grouped together in one cluster but accessions from India and Nepal were grouped together in another cluster. However, in both the species accessions from Turkey did not group together and fell in different clusters. CONCLUSIONS: Based on genetic similarity indices, higher diversity was observed in Trigonella caerulea as compared to Trigonella foenum-graecum. The genetic similarity matrices generated by ISSR and RAPD markers in both species were highly correlated (r = 0.78 at p = 0.001 for Trigonella foenum-graecum and r = 0.98 at p = 0.001 for Trigonella caerulea) indicating congruence between these two systems. Implications of these observations in the analysis of genetic diversity and in supporting the possible Center of Origin and/or Diversity for Trigonella are discussed.

DNA, Plant↗

Spinal serotonergic system is partially involved in antinociception induced by Trigonella foenum-graecum (TFG) leaf extract.

It has been reported that Trigonella foenum-graecum (TFG) extract exerts analgesic, anti-inflammatory and anti-pyretic effects in different experimental models. The major objective of this paper was to investigate the site and mechanism of the analgesia induced by Trigonella foenum-graecum extract. We studied the analgesic effects of different doses of Trigonella foenum-graecum extract after i.p., i.t. and i.c.v. administration in formalin test, using male NMRI rats (200-250 g). Trigonella foenum-graecum extract showed analgesic effects in i.p. (1 g/kg) and i.t. (0.5, 1, and 2 mg/rat) (P < 0.05 in all groups) but not in i.c.v. (1 and 3 mg/rat) administrations. Based on the similarities between the effects of Trigonella foenum-graecum extract with those of nonsteroidal anti-inflammatory drugs (NSAIDs) and the role of 5-HT system in analgesic effects of NSAIDs, we tried to investigate the role of spinal 5-HT system in analgesic effects of Trigonella foenum-graecum extract. After lesioning of spinal 5-HT system by 5,7-dihydroxytryptamine (5,7-DHT), it was shown that the analgesic effect of Trigonella foenum-graecum extract (0.5 and 3 mg/rat) in the second phase of formalin test, was abolished completely and reduced relatively after using a low-dose (0.5 mg/rat) and a high-dose (3 mg/rat), respectively (P < 0.05). So, the antinociception partially remained (P < 0.05) after using the latter dose. Meanwhile, administration of naloxone (2mg/kg, i.p.) had no effect on the Trigonella foenum-graecum extract (1 g/kg, i.p.) analgesia. In conclusion, this study confirms the central action of Trigonella foenum-graecum extract and that spinal 5-HT system is partially involved in the analgesia induced by it in the second phase of formalin test and also indicates for co-existence of other analgesic mechanism(s).

Analgesics↗

Hypoglycaemic and antihyperglycaemic effects of Trigonella foenum-graecum leaf in normal and alloxan induced diabetic rats.

The aqueous and alcoholic extracts of Trigonella foenum-graecum leaf were tested for hypoglycaemic activity in normal and alloxan-diabetic rats. Graded amounts (0.06, 0.2, 0.5, 1 g/kg, i.p. and 1, 2, 8 g/kg, p.o.) of the aqueous extract of Trigonella foenum-graecum leaf when given to both normal and alloxan-diabetic rats, a significant reduction of blood glucose concentration was noticed. On the other hand ethanolic extract of Trigonella foenum-graecum leaf produced no reduction in blood glucose concentration in normal rats but intra-peritoneal administration of 0.8 g/kg of the ethanolic leaf extract to diabetic rats produced a significant reduction of blood glucose concentration (p < 0.02) at 2 and 24 h only. Intraperitoneal and oral acute toxicity (LD50) and target organ effects were studied for the aqueous extract of Trigonella leaf in mice. LD50 of i.p. and oral administration were 1.9 and 10 g/kg respectively. The main organ affected after i.p. administration of the aqueous extract was the liver while oral administration of the aqueous extract of Trigonella did not produce any sign of organ damage. These results suggest that the aqueous extract of Trigonella foenum-graecum leaves given both orally and intraperitoneally possesses a hypoglycaemic effect in normoglycaemic and alloxan induced hyperglycaemic rats.

Administration, Oral↗

[Effect of vitamin E and protein quality on the hemolytic effect of Trigonella sapogenins in rats].

The influence of vitamin E (9, 90, 450 mg dl-alpha-tocopherol acetate/100 g diet), protein quality and temperature (70 degrees C for 10 min) on the sapogenin-induced hemolysis was studied in rats fed with diets containing different amounts of fenugreek-seed seedflour. Significant increased hemolysis rates were found in rats on diets with the following protein components: 1/3 Trigonella + 2/3 Cornflour (69.7 +/- 11.6% hemolysis), 100% Trigonella (21.0 +/- 8.6% hemolysis), 1/3 Trigonella + 2/3 Casein (14.1 +/- 10.7% hemolysis). The diets of these groups of animals contained raw Trigonella seeds and 9 mg vitamin E/100 g diet, which covers the requirements of rats on this vitamin. With this tocopherol supplement the applied heat treatment had no influence on the sapogenin effects on the erythrocytes. In the groups with the 10th fold and 50th fold of the vitamin E supplementation no significant hemolysis rate was observed. From these results we conclude that when fenugreek seeds are to be used in animal nutrition, a qualitative good protein supply as well as a sufficient intake of vitamin E should be considered in order to compensate the hemolytic effect of Trigonella sapogenins.

Animals↗

Restoration of ultrastructural and biochemical changes in alloxan-induced diabetic rat sciatic nerve on treatment with Na3VO4 and Trigonella--a promising antidiabetic agent.

Vanadium has been reported to have broad pharmacological activity both in vitro and in vivo. Vanadium compound, sodium orthovanadate, Na3VO4, is well known for its hypoglycaemic effects. However, Na3VO4 exerts these effects at relatively high doses (0.6 mg/ml) and exhibit several toxic effects. In the present study lower doses of Na3VO4 (0.2 mg/ml) are combined with Trigonella foenum graecum seed powder (TSP), another hypoglycaemic agent, to reduce its toxicity without compromising its antidiabetic potential. The efficacy of the lower doses of Na3VO4 has been investigated in restoring the altered glucose metabolism and histological structure in the sciatic nerves in 21 and 60 days alloxan diabetic rats. A portion of the glucose was found to be channelled from the normal glycolytic route to polyol pathway, evident by the reduced hexokinase activity and increased polyol pathway enzymes aldose reductase and sorbitol dehydrogenase activity causing accumulation of sorbitol and fructose in diabetic conditions. Ultrastructural observation of the sciatic nerve showed extensive demylination and axonal loss after eight weeks of diabetes induction. Blood glucose levels increased in diabetic rats were normalized with the lower dose of vanadium and Trigonella treatment. The treatment of the diabetic rats with vanadium and Trigonella prevented the activation of the polyol pathway and sugar accumulations. The sciatic nerves were also protected against the structural abnormalities found in diabetes with Trigonella foenum graecum as well as Na3VO4. Results suggest that lower doses of Na3VO4 may be used in combination with TSP as an efficient antidiabetic agent to effectively control the long-term complications of diabetes in tissues like peripheral nerve.

Aldehyde Reductase↗

Lower doses of vanadate in combination with trigonella restore altered carbohydrate metabolism and antioxidant status in alloxan-diabetic rats.

BACKGROUND: Vanadate treatment to diabetic rats has been reported to correct the altered carbohydrate metabolism and antioxidant status. However, vanadate exerts these effects at relatively high doses and several toxic effects are produced. We used low doses of vanadate in combination with Trigonella foenum graecum seed powder (TSP) and evaluated their effect on the enzyme changes in diabetic rats. METHODS: Alloxan-diabetic rats were treated separately with insulin, vanadate (0.6 mg/ml), TSP and a combined dose of Vanadate (0.2 mg/ml) and TSP for 21 days. At the end of the experimental period, blood glucose levels and activities of pyruvate kinase (PK), phosphoenolpyruvate carboxykinase (PEPCK), glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD) and catalase (CAT) were measured in cytosolic fraction in the liver and kidney. RESULTS: Blood glucose levels increased markedly in diabetic rats. Treatment with antidiabetic compounds resulted in the reduction of glucose levels. Rats treated with combined dose of vanadate and trigonella had glucose levels comparable to control ones. Similar results were obtained with the activities of PK, PEPCK, SOD, GPx, GR, and CAT in liver and kidney of diabetic rats. Combined dose of vanadate and Trigonella was found to be most effective in correcting these alterations. CONCLUSIONS: Lower doses of vanadate could be used in combination with TSP to effectively counter diabetic alterations without any toxic side effects.

Animals↗

The combined effects of Trigonella and Allium extracts in the regulation of hyperthyroidism in rats.

The combined effects of Trigonella foenum-graecum and Allium sativum extracts were evaluated for their ameliorative potential in the L-thyroxine-induced hyperthyroidic rat model to contribute to an understanding of interaction between the two extracts. The investigation was carried out using two different doses. A comparison was made with the response of individual plant extracts at the previously studied effective dose in adult Wistar rats rendered hyperthyroidic by daily injections of L-thyroxine (300 microg/kg body wt., s.c.). Propylthiouracil (PTU), an antithyroid drug, was used as a reference compound. Alterations in serum triiodothyronine (T3), thyroxine (T4), glucose, hepatic glucose-6-phosphatase (G-6-Pase) and oxygen consumption were studied as end parameters. Superoxide dismutase (SOD), catalase (CAT) activities, lipid peroxidation (LPO) and reduced glutathione (GSH) were examined to reveal any toxic effects of the drugs. The combined effects of Trigonella and Allium at 200 and 500 mg/kg body wt. respectively, were equipotent as compared to the individual extracts in lowering the serum concentrations of T3 and T4 in hyperthyroidic rats. Our findings reveal that some plant extracts in combination may not always prove to be synergistic. It is therefore suggested that Trigonella foenum-graecum and Allium sativum extracts may be used individually and not together in the regulation of hyperthyroidism.

Animals↗

Evaluation of the oral hypoglycaemic effect of Trigonella foenum-graecum L. (methi) in normal mice.

Trigonella foenum-graecum (Fenugreek) (Leguminosae) is employed as a herbal medicine. Its seeds are known for their carminative, tonic and antidiabetic effects. A curative dose of Trigonella foenum-graecum also produces antiulcer action. In this study we have investigated the hypoglycaemic activities of the aqueous extract of the seeds Trigonella foenum-graecum in normal mice using oral route of adminstration. The methanolic extract administered through the same route produced hypoglycaemic effect only at the dose of 1 g/kg body weight. The aqueous extract is under further investigation to determine the chemical structure of the active component. The presence of hypoglycaemic activity in aqueous and methanolic extract indicates that the active compounds are polar in nature.

Administration, Oral↗

Trigonella foenum graecum (fenugreek) seed extract as an antineoplastic agent.

The antineoplastic effect of Trigonella foenum graecum seed extract has been evaluated in the Ehrlich ascites carcinoma (EAC) model in Balb-C mice. Intra-peritoneal administration of the alcohol extract of the seed both before and after inoculation of EAC cell in mice produced more than 70% inhibition of tumour cell growth with respect to the control. Treatment with the extract was found to enhance both the peritoneal exudate cell and macrophage cell counts. The extract also produced a significant antiinflammatory effect. We report here the antiinflammatory and antineoplastic effects, of Trigonella foenum graecum seed extract.

Animals↗

Amelioration of altered antioxidant status and membrane linked functions by vanadium and Trigonella in alloxan diabetic rat brains.

Trigonella foenum graecum seed powder (TSP) and sodium orthovanadate (SOV) have been reported to have antidiabetic effects. However, SOV exerts hypoglycemic effects at relatively high doses with several toxic effects. We used low doses of vanadate in combination with TSP and evaluated their antidiabetic effects on anti-oxidant enzymes and membrane-linked functions in diabetic rat brains. In rats, diabetes was induced by alloxan monohydrate (15 mg/100 g body wt.) and they were treated with 2 IU insulin, 0.6 mg/ml SOV, 5% TSP and a combination of 0.2 mg/ml SOV with 5% TSP for 21 days. Blood glucose levels, activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), Na+/K+ ATPase, membrane lipid peroxidation and fluidity were determined in different fractions of whole brain after 21 days of treatment. Diabetic rats showed high blood glucose (P less than 0.001), decreased activities of SOD, catalase and Na+/K+ ATPase (P less than 0.01, P less than 0.001 and P less than 0.01), increased levels of GPx and MDA (P less than 0.01 and P less than 0.001) and decreased membrane fluidity (P less than 0.01). Treatment with different antidiabetic compounds restored the above-altered parameters. Combined dose of Trigonella and vanadate was found to be the most effective treatment in normalizing these alterations. Lower doses of vanadate could be used in combination with TSP to effectively counter diabetic alterations without any toxic effects.

Animals↗

Breeding system in a population of Trigonella balansae (Leguminosae).

BACKGROUND AND AIMS: Although some taxonomic studies in the genus Trigonella have been conducted, there has been no concerted effort to study the breeding system. This paper examines the floral structure and pollination system in a population of T. balansae, an annual pasture legume. METHODS: Floral morphology, hand and vector pollination, stigma receptivity, pollen tube growth, using scanning electron and fluorescence microscopy, were conducted. KEY RESULTS: Measurements of floral structure from before to after anthesis indicates an inability for T. balansae to self-pollinate and a requirement for an external vector to effectively transfer pollen from the anthers onto the stigmas of this species. Seed set can be obtained by hand or honeybee manipulation of T. balansae flowers. CONCLUSIONS: Trigonella balansae is a self-compatible species, but which requires vectors such as honeybees to bring about pollination.

Flowers↗

In vivo effect of Trigonella foenum graecum on the expression of pyruvate kinase, phosphoenolpyruvate carboxykinase, and distribution of glucose transporter (GLUT4) in alloxan-diabetic rats.

Plasma glucose levels are maintained by a precise balance between glucose production and its use. Liver pyruvate kinase (PK) and phosphoenolpyruvate carboxykinase (PEPCK), 2 key enzymes of glycolysis and gluconeogenesis, respectively, play a crucial role in this glucose homeostasis along with skeletal muscle glucose transporter (GLUT4). In the diabetic state, this balance is disturbed owing to the absence of insulin, the principal factor controlling this regulation. In the present study, alloxan-diabetic animals having high glucose levels of more than 300 mmol/L have been taken and the administration of Trigonella seed powder (TSP) to the diabetic animals was assessed for its effect on the expression of PK and PEPCK in liver and GLUT4 distribution in skeletal muscle of alloxan-diabetic rats. TSP treatment to the diabetic animals resulted in a marked decrease in the plasma glucose levels. Trigonella treatment partially restored the altered expression of PK and PEPCK. TSP treatment also corrected the alterations in the distribution of GLUT4 in the skeletal muscle.

Animals↗

[Studies on flavonoids from Fenugreek (Trigonella foenumgraecum L.)].

OBJECTIVE: To study the flavonoids in fenugreek (Trigonella foenumgraecum) produced in China. METHOD: The flavonoids were isolated with silica gel, polyamide and Sephadex LH-20 chromatography, and their structures were identified by physical, chemical properties and spectral analysis. RESULT: Five flavonoid compounds were isolated from fenugreek seeds and identified as vitexin, tricin, naringenin, quercetin and tricin-7-O-beta-D-glucopyranoside. CONCLUSION: Three flavonoids, tricin, naringenin and tricin-7-O-beta-D-glucopyranoside, were isolated from fenugreek as well as from the plants of Trigonella for the first time.

Apigenin↗

Antagonism of cadmium and alloxan-induced hyperglycemia in rats by Trigonella foenum graecum.

This study was stimulated by popular belief that Trigonella foenum graecum has antidiabetic activity in which the hypoglycemic effect has been confirmed by several investigators. However, the mode of action appears to be unclear. To gain some insight, the stems and leaves of the plant as well as the known active seeds were extracted at room temperature (ca 20 C) for three days with water or acetone and by soxhlet 70 C, and tested by oral administration to rats. Acetone and CC14 extracts before use, were evaporated below 30 C under vacuo, and the residue dissolved in distilled water containing Tween 80. Hypoglycemic activity of these extracts were tested on 20 hour fasted normal, alloxan and cadmium treated rats. The latter has been shown to cause hyperglycemia by releasing epinephrine in intact rats and inhibiting insulin release in the isolated perfused rat pancreas. Results showed that with seeds the CC14, soxhlet acetone extracts were inactive in normal animals as were the water and acetone extracts of stems and leaves. These observations may be compared with room temperature acetone extraction of seeds which exhibited what appeared to be dose related hypoglycemic effects. The hyperglycemia induced by cadmium or alloxan was antagonized by room temperature acetone seed or stem and leaves extracts. Tentative interpretation of the above results, are that Trigonella acetone extract appears to act, at least in part, at the cellular level to produce its hypoglycemic effects on normal rats or those as treated with cadmium or alloxan.

Acetone↗

Protective effect of fenugreek (Trigonella foenum graecum) seeds in experimental ethanol toxicity.

The study investigates the effect of aqueous extract of fenugreek seeds (Trigonella foenum graecum) on lipid peroxidation and antioxidant status in experimental ethanol toxicity in rats. The ability of the seed extract to prevent iron-induced lipid peroxidation in vitro was also investigated. Ethanol feeding for 60 days resulted in significant increases in the activities of serum aspartate transaminase, alanine transaminase and alkaline phosphatase. The levels of serum lipid hydroperoxides and thiobarbituric acid reactive substances in liver and brain were also significantly elevated. Significantly lower activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase and glutathione reductase were observed in liver and brain accompanied by depletion in glutathione, ascorbic acid and alpha-tocopherol concentrations. Activity of Ca(2+) ATPase in brain was significantly lowered. Simultaneous administration of aqueous extract of fenugreek seeds with ethanol prevented the enzymatic leakage and the rise in lipid peroxidation and enhanced the antioxidant potential. The seeds exhibited appreciable antioxidant property in vitro which was comparable with that of reduced glutathione and alpha-tocopherol. Further, histopathological examination of liver and brain revealed that, aqueous extract of fenugreek seeds could offer a significant protection against ethanol toxicity.

Alanine Transaminase↗

Nematicidal activity of Trigonella foenum-graecum L.

The aqueous, methanol and chloroform extracts of Trigonella foenum-graecum caused significant (p < 0.05) mortality of Meloidogyne javanica larvae. The methanol soluble fraction eluted from pure distilled water showed the highest (>92%) nematicidal activity compared with the fractions eluted from pure methanol and different ratios of chloroform and methanol indicate that the nematicidal compound was polar in nature.

Animals↗