COMPARATIVE EFFECT OF FENUGREEK (TRIGONELLA FOENUM GRACEUM) AND GINGER (ZINGIBER OFFICINALE ROSCOE) ON DIABETIC ADULT MALE ALBINO RATS

Document Type : Original Article

Author

Medical Physiology Department, Faculty of Medicine, Al-Azhar University

Abstract

Background: Diabetes mellitus (DM) is a chronic disease and it is one of the major health problems due to its high prevalence, chronic nature and high risk of chronic complications. Currently available therapeutic options for DM have little or no effect on lipid profile, have various adverse effects and are far from satisfactory as far as long-term complications are concerned.
Objectives: The present study was designed to evaluate and compare the efficacy of fenugreek and ginger on lipid profile, blood sugar and glycated hemoglobin (HbA1c) levels in diabetic adult male albino rats.
Materials and Methods: Thirty two adult male albino rats of local strain were used and divided into four equal groups: group I (control group) was given distilled water orally, group II (diabetic group), group III (diabetic treated with fenugreek extract 1g/kg body weight, single oral daily dose for 12 weeks), and group IV (diabetic treated with ginger extract (0.5g/kg body weight in a single dose orally daily for 12 weeks). Diabetes induced by a single intraperitoneal injection of alloxan (150 mg/kg body weight) in freshly prepared physiological saline.
Results: Diabetic animals showed marked significant elevations in TG, TC and LDL, accompanied with marked decline in HDL relative to the corresponding controls. Treatment of diabetic rats with fenugreek and ginger improved the sera lipids profile as shown by the significant reduction in the values of TC, TC and LDL associated with marked elevation of HDL without significant differences between both treated groups. A significant decrease in the levels of serum insulin accompanied with marked significant elevation in the levels of blood glucose and HbA1c were recorded in diabetic rats when compared to the control rats .Marked recovery in insulin, glucose and HbA1c levels was recorded in diabetic animals post treated with fenugreek and ginger without significant differences between each other.
Conclusion: Fenugreek and ginger showed significant hypolipidemic and hypoglycemic effects in diabetic rats.

Keywords


COMPARATIVE EFFECT OF FENUGREEK (TRIGONELLA FOENUM GRACEUM) AND GINGER (ZINGIBER OFFICINALE ROSCOE) ON DIABETIC ADULT MALE ALBINO RATS

By

 

Mohammad Abulhasan Zoair

Medical Physiology Department, Faculty of Medicine, Al-Azhar University

ABSTRACT

Background: Diabetes mellitus (DM) is a chronic disease and it is one of the major health problems due to its high prevalence, chronic nature and high risk of chronic complications. Currently available therapeutic options for DM have little or no effect on lipid profile, have various adverse effects and are far from satisfactory as far as long-term complications are concerned.

Objectives: The present study was designed to evaluate and compare the efficacy of fenugreek and ginger on lipid profile, blood sugar and glycated hemoglobin (HbA1c) levels in diabetic adult male albino rats.

Materials and Methods: Thirty two adult male albino rats of local strain were used and divided into four equal groups: group I (control group) was given distilled water orally, group II (diabetic group), group III (diabetic treated with fenugreek extract 1g/kg body weight, single oral daily dose for 12 weeks), and group IV (diabetic treated with ginger extract (0.5g/kg body weight in a single dose orally daily for 12 weeks). Diabetes induced by a single intraperitoneal injection of alloxan (150 mg/kg body weight) in freshly prepared physiological saline.

Results: Diabetic animals showed marked significant elevations in TG, TC and LDL, accompanied with marked decline in HDL relative to the corresponding controls. Treatment of diabetic rats with fenugreek and ginger improved the sera lipids profile as shown by the significant reduction in the values of TC, TC and LDL associated with marked elevation of HDL without significant differences between both treated groups. A significant decrease in the levels of serum insulin accompanied with marked significant elevation in the levels of blood glucose and HbA1c were recorded in diabetic rats when compared to the control rats .Marked recovery in insulin, glucose and HbA1c levels was recorded in diabetic animals post treated with fenugreek and ginger without significant differences between each other.

Conclusion: Fenugreek and ginger showed significant hypolipidemic and hypoglycemic effects in diabetic rats.

Key words: Fenugreek, Ginger, Diabetes, Lipid Profile.

 

 

INTRODUCTION

     Diabetes mellitus is a chronic disease and it is one of the major health problems due to its high prevalence, chronic nature and high risk of chronic complications. It is commonly associated with dyslipidemia, which is one of the major risk factors of coronary heart disease (CHD) in diabetic patients (Amandeep et al., 2016).

     Fenugreek (Trigonella foenum graceum) is one of the oldest herb used for medicinal purposes and its history dates back to Egyptian civilization (Ansari and Ansari, 2011). Fenugreek seeds contain alkaloids, including trigonelline, gentianine and carpaine compounds.

     Ginger has a long history of use as herbal medicine to treat a variety of diseases since it can scavenge superoxide anion and hydroxyl radicals (Elshater et al., 2009).

     The present study was designed to evaluate and emphasize the efficacy of fenugreek seed and ginger on lipid profile, blood sugar and glycated hemoglobin (HbA1c) levels in diabetic adult male albino rats.

MATERIALS AND METHODS

     Thirty two adult male albino rats of a local strain weighing approximately 140-160 g, were purchased from Animal House Centre, Al-Azhar University, Cairo, Egypt, were housed in clear plastic cages (40x 30x 35 cmm, 4 animals /cage) with wood chips as bedding and given a standard pellet rodent diet, in addition to water ad libitum. The rats were maintained under standard laboratory conditions at 25±2oC and normal light/dark cycle, and kept for ten days before experiment for adaptation.

Induction of diabetes: The animals were rendered diabetics by a single intraperitoneal injection of alloxan (150 mg/kg body weight) in a freshly prepared physiological saline. Diabetic state of animals was monitored for its stability for seven successive days after alloxan treatment. On day eight of alloxan injection, only animals with fasting blood glucose levels >200 mg/dl were selected as diabetic rats for the current experiment (Szkudelski, 2001).

Plant Materials and Preparation: The seeds of Fenugreek (Trigonella foenum graecum), and raw dry ginger rhizomes (Zingiber officinale Roscoe) were obtained from a local herbal commercial market. The fenugreek seeds were dried then homogenized to fine powder. The extract of fenugreek was prepared by boiling of 30 g of fenugreek with 150 ml water for 5 minutes, left cool to room temperature, then filtered and stored at -20ºC till used.  The Fenugreek aqueous extract solution was given by gastric gavage (single oral daily dose 1g/kg body weight) (Marzouk et al., 2013).  Also, ginger  was grinded to fine powder and extracted with aqueous solution by boiling of 15 g of ginger with 150 ml water for 5 minutes, left cool to room temperature, then filtered and stored at -20ºC till used and given by gastric gavage (single oral daily dose 0.5 g/kg body weight) (Akhani et al., 2005) .

Experimental Design: rats were divided into four equal groups: group I (control group) given distilled water orally, group II (diabetic group), group III (diabetic treated with fenugreek 1g/kg body weight, single oral daily dose for 12 weeks), and group IV (diabetic treated with ginger 0.5g/kg body weight, single oral daily dose for 12 weeks).

Collection of blood and estimation of biochemical parameters: At the end of experiment, blood samples were collected after overnight fasting rats in centrifuge tubes by retro-orbital puncture under isofluren anesthesia. Blood samples were used in separation of sera by centrifugation at 4000 rpm for 10 min at 4°C, and immediately stored at -20°C for further analysis of biochemical parameters.

     Serum total cholesterol (Henry et al., 1997), triglycerides (Fossati and Principe, 1982), and HDL (Burstein and Scholnick, 1972) were estimated colorimetrically, while LDL was calculated by applying the Friedwald's equation (Friedewald et al., 1972).

     Serum glucose was estimated according to the method of (Trinder, 1969). Serum insulin level was measured by an enzyme immunoassay kit (SPI-Bio société de pharmacologieetd'lmmunoloie-Bio, France), while values of HbA1c level were determined by ion exchange resin method (Bunn, 1981).

Statistical analysis:

     The results were expressed as mean ± SE, and the statistical significance was evaluated by one way analysis of variance (ANOVA) followed by Duncan post Hoc test using the SPSS/17.0 software. Values were considered statistically significant at P ≤ 0.05.


RESULTS

    

 

     Diabetic animals showed marked significant elevations in TG, TC and LDL accompanied with marked decline in HDL relative to the corresponding controls. Treatment of diabetic rats with fenugreek and ginger improved the sera lipids profile as shown by the significant reduction in the values of TG, TC and LDL associated with marked elevation of HDL without significant differences between both treated groups (Table 1).

 

 

Table (1): Effect of Fenugreek and Ginger on serum lipid profiles in diabetic rats (mean ± SE)

HDL-C

(mg/dl)

 

LDL-C

(mg/dl)

Cholesterol

(mg/dl)

Triglyceride

(mg/dl)

Parameters

 

Groups

 

45.80

±1.11

 

 

17.76

±0.89

86.51

± 1.55

113.32

± 4.02

GI

 

30.76a

±0.57

 

51.16 a

± 1.77

144.22a

± 3.56

203.24a

± 0.11

GII

 

40.60bc

±1.06

 

22.96bc

± 1.29

94.67bc

± 2.14

164.15bc

± 3.19

GIII

39.97bc ±1.01

 

32.88bc

± 4.03

98.35bc

± 4.66

173.33bc

± 0.72

GIV

GI: control group         

GII: diabetic group  

GIII: diabetic group treated with fenugreek                    

GIV: diabetic group treated with ginger    

a: Significant as compared with normal control (I)

b: Significant  as compared with the diabetic control (II)

c: insignificant GIII and GIV compared with each other 

 

 

     Significant decrease in the levels of serum insulin accompanied with marked significant elevation in the levels of blood glucose and HbA1c were recorded in diabetic rats when compared to the control rats. Marked recovery in insulin, glucose and HbA1c levels was recorded in diabetic animals post treated with fenugreek and ginger without significant differences between each other (Table 2).


Table (2): Effect of Fenugreek and Ginger on serum glucose, insulin and HbA1c in diabetic rats (mean ± SE)

HbA1c

 %

 

  Insulin

(mg/ml)

Glucose

(mg/dl)

Parameters

 

Groups

 

 

5.37±1.83

 

 

 

       

 

26.47 ± 1.31

76.10 ± 1.31

GI

7.19a  ± 1.30

 

20.11a ± 0.93

210.31a ±5.13

GII

5.40bc ± 1.16

 

23.23bc ± 0.17

98.44bc ± 4.81

GIII

5.39bc ± 1.76

 

25.89bc ± 0.14

94.82bc ± 2.16

GIV

GI: control group         

GII: diabetic group   

GIII: diabetic group treated with fenugreek                     

GIV: diabetic group treated with ginger    

a: Significant as compared with normal control (I)

b: Significant  as compared with the diabetic control (II)

c: insignificant GIII and GIV compared with each other 


DISCUSSION

     The current study revealed high prevalence of hypercholesterolemia, hypertriglyceridemia, high LDL and low HDL levels in diabetic rats compared to control group which are risk factors in patients with diabetes (Amandeep et al., 2016)

     Fenugreek-treated group showed significant decrease in total cholesterol and triglycerides levels in comparison to the diabetic group. These results agreed with Reddy and Srinivasan. (2011) who showed that fenugreek leads to delayed onset of cholesterol crystallization.

     Abedinzade et al. (2013) stated that the hypolipidemic action of fenugreek extract caused by delaying lipids and carbohydrates absorption as a result of bioactive fibers existing in fenugreek seeds. Also, fenugreek seeds decrease ApoB and lecithin catalase enzyme as well as phospholipids level which play an important role in cholesterol synthesis (Marzouk et al., 2013).

     On the other hand, ginger-treated group showed also a significant decrease in total cholesterol and triglycerides levels compared to diabetic group, but insignificant with fenugreek-treated group. These results agreed with Lebda et al.( 2012) and Kalaiselvi et al. (2015) who attributed the hypolipidemic effect of ginger to stimulating the conversion of cholesterol to bile acids and inhibition pancreatic lipase enzyme, as well as inhibition of cellular cholesterol biosynthesis.

     Fenugreek-treated and ginger-treated groups showed a significant decrease in LDL level, while HDL level increased compared to diabetic group without significant differences between each others. The present results agreed with Shrivastava et al. (2009) and Abedinzade et al. (2013) that fenugreek decreases cholesterol content of the bile by increasing bile secretion. Results agreed with Lebda et al. (2012) and (Kalaiselvi et al., 2015) that ginger decreases level of LDL-receptor and inhibition of hepatic fatty acid synthesis.

     Ginger and fenugreek treated groups showed significant decrease in serum glucose level and HbA1c in comparison to diabetic group without significant differences between each others. Results of fenugreek treated group agreed with Manmeet et al. (2016). The cause of reduction was attributed to slow release of carbohydrate and increase the soluble fiber, which helped lower blood sugar by slowing down digestion and absorption of carbohydrates. Fenugreek seeds contain galactose and mannose which are associated with reduced hyperglycemia and hypercholesterolemia, the seeds also has an insulinotropic effect. 

     Abedinzade et al. (2013) indicated that fenugreek extract causes control of intestinal absorption of glucose under lab conditions.

     Ginger group results agreed with Kazeem et al. (2015) who stated that ginger might be exerting insulin-like effect on peripheral tissues by promoting glucose uptake, inhibiting hepatic gluconeogenesis or increased entrance of glucose into the muscle and adipose tissues through the stimulation of glucose transporter gene expression "Glut-4", and stimulation of a regeneration process of the remaining β-cells in addition to gingerols inhibit α–glucosidase and α–amylase enzymes. Jafri et al. (2011) also showed that oral administration of ginger extract with daily dose of 500 mg/Kg for 6 weeks in alloxan-diabetic rats causes decrease in blood glucose level.

      In parallel, serum insulin level significantly decreased in the diabetic rats compared with normal control, and significantly increased in fenugreek-treated and ginger-treated groups compared with diabetic group. Insulin deficiency ultimately results in increased production of glucose by the liver, and decreased utilization of glucose in peripheral tissues (Giugliano et al., 2008). This indicated that changes in insulin may bring about changes in hepatic glycogen content and lead to the regulatory effect of fenugreek and ginger on glucose metabolism. Pandit et al.( 2010) indicated that soluble dietary fiber of fenugreek foenum graecum seed exerts antidiabetic effects mediated through inhibition of carbohydrate digestion and absorption, and enhancement of peripheral insulin action.

CONCLUSION

     Fenugreek and ginger showed significant hypolipidemic and hypoglycemic effects in diabetic rats, but without significant differences between each other. So, administration of medicinal plants such as fenugreek and ginger could ameliorate the disturbances caused by diabetes without the disadvantages of chemically synthetic drugs.

REFERENCES

1. Abedinzade M, Nikokar I, Nasri S, Omidi JM and Nursabaghi F. (2013):  Effect of hexanic and alcoholic extract of fenugreek seed in male diabetic rats. Zahedan J. Res. Med. Sci. (ZJRMS), 15: 50-53.

2. Akhani SP, Vishwakarma Sl and Goyal Rk. (2005): Antidiabetic activity of zingiber officinale roscoe in streptozotocin-induced non-insulin dependent diabetic rats. Indain J. Pharm. Sci., 67 (5): 553-557.

3.  Amandeep S, Jaswant R and Devinder SM. (2016): Comparative evaluation of glipizide and fenugreek (Trigonella foenumgraecum) seeds as monotherapy and combination therapy on glycaemic control and lipid profile in patients with type 2 diabetes mellitus. International Journal of Basic & Clinical Pharmacology, 5 (3): 942-450.

4. Ansari R and Ansari S. (2011): Effectiveness of fenugreek for lowering the hemoglobin (HbA1c) in the patients with self-management of type 2 diabetes: A randomized control trial. Medical complication of type-2 diabetes. Dr. Collen Croniger (Ed). Pbl. InTech Europe: England: pp. 339-412.

5. Bunn HF. (1981): Evaluation of glycosylated haemoglobin diabetic patients. Diabetes, 30 (7):613-617.

6. Burstein RF and Scholnick VS. (1972):  Biochemistry and methodology of lipids. J Lipid Res., 25: 375-382.

7. Elshater AEA, Muhammad MAS and Mahrous MA M. (2009): Effect of ginger extract consumption on levels of blood glucose, lipid profile and kidney functions in Alloxan induced-diabetic rats. Egypt. Acad. J. Biology. Sci., 2: 153-162.

8.  Fauci AS, Braunwald E, Kasper DL, Stephen LH, Dan LL, Jameson JL and Loscalzo J. (2012): Harrison’s Principles of Internal Medicine.18th editors. pbl. New York: McGraw Hill: pp. 2968-3002.

9. Fossati P and Principe L. (1982): Serum triglycerides determined calorimetrically with an enzyme that produces hydrogen peroxide. Clin.Chem., 28: 2077-2080.

10. Friedewald WT, Levey RI and Fredrickson DS. (1972): Estimation of the concentration of low density lipoprotein cholesterol in plasma without use of the preparative ultracentrifuge. Clin. Chem., 18:499-502.

11. Giugliano D, Ceriello A and Esposito K. (2008): Glucose metabolism and hyperglycemia. Am J Clin Nutr., 87: 217-222.

12. Henry RJ, Cannon DC and Winkelman JW. (1997): Clinical Chemistry Principles and Tetchiness, pbl. New York: Harper and Row: p.1440.

13. Jafri SA, Abass S and Qasim M. (2011): Hypoglycemic effect of ginger (zingiber officinale) in alloxan induced diabetic rats (Rattusn norvagicus). Pak. Vet. J., 31: 160-162.

14. Kalaiselvi A, Reddy GA and Ramalingam V. (2015): Ameliorating Effect of Ginger Extract (Zingiber officinale Roscoe) on Liver Marker Enzymes, Lipid Profile in Aluminum chloride Induced Male Rats. IJPSDR, 7: 52-58.

15. Kazeem MI, Akanji MA, Yakubu MT and Ashafa AO. (2015): Antiglycation and Hypolipidemic Effects of Polyphenols from Zingiber officinale Roscoe (Zingiberaceae) in Streptozotocin-Induced Diabetic Rats. Tropical J. of Pharmaceutical Res., 14: 55-61.

16. Lebda MA, Taha NM,  Korshom MA, Mandour AA and El-Morshedy AM.( 2012): Biochemical effect of ginger on some blood and liver parameters in male Newzeland rabbits. Online J. Animal and Feed Research, 2: 197- 202.

17. Manmeet K, Narinder S, Geeta S and Davinder S. (2016): To study the efficacy and tolerability of fenugreek seed powder as add-on therapy with metformin in patients of type-2 diabetes mellitus. International Journal of Basic & Clinical Pharmacology, 5 (2): 378-383.

18. Marzouk M, Soliman AM and Omar TM. (2013): Hypoglycemic and antioxidative effects of fenugreek and termis seeds powder in streptozotocin-diabetic rats. European Review for Medical and Pharmacological Sciences, 17: 559-565.

19. Pandit R, Phadke A and Jagtap A. (2010): Antidiabetic effect of Ficus religiosa extract in streptozotocin induced diabetic rats. J Ethnopharmacol., 128: 462-466.

20. Reddy RR and Srinivasan K. (2011): Effect of dietary fenugreek seeds on biliary proteins that influence nucleation of cholesterol crystals in bile. ELSEVIER Steroids, 76: 455-463.

21. Shrivastava R, Solanki SS, Tomar V, Garud N, Garud A, Kannojia P and Jain N. (2009):  Comparative Evaluation of Polyherbal Combination for Hypolipidemic Activity. International Journal of Pharmaceutical Sciences and Drug Research, 17: 9-12.

22. Szkudelski T. (2001):  The mechanism of Alloxan and alloxan Action in β cells of the rats pancreas. Physiol Res., 50: 536- 546.

23. Trinder P. (1969): Determination of glucose in blood using glucose oxidase with an alternative oxygen acceptor. Ann. Clin. Biochem., 6: 24-33.

 


مقارنة تأثیر نباتی الحلبة والزنجبیل على  ذکورالجرذان البیضاء البالغة المصابة بمرض البوال السکری

محمد أبوالحسن زعیر

قسم الفسیولوجیا الطبیة - کلیة الطب - جامعة الأزهر

خلفیة البحث : مرض البوال السکری هو أحد المشاکل الصحیة الرئیسیة فى العالم بسبب کثرة شیوعه وطبیعته المزمنة وکثرة مخاطره. والعلاج العشبی یکتسب شعبیة وإنتشاراً فی علاج مرض البوال السکری ومضاعفاته.

الهدف من البحث: تقییم ومقارنة تأثیر نباتی الحلبة والزنجبیل على مستوی دلائل الدهون والجلوکوز والهیموجلوبین السکری فى الدم فی ذکورالجرذان البیضاء البالغة المصابة بمرض البوال السکری.

موادوطرق البحث: تم تقسیم إثنین وثلاثین جرذاً إلی أربعة مجموعات متساویة [المجموعة الضابطة ـ مجموعة البوال السکری ـ مجموعة البوال السکری المعالجة بالحلبة - مجموعة البوال السکری المعالجة بالزنجبیل] .

       وقد تم إحداث البوال السکری باستخدام جرعة واحدة من الألوکسان بالغشاء البروتینى  [150 مجم / کجم] ، کما تم إعطاء مستخلص نبات الحلبة  بالفم مرة واحدة یومیاً لمدة 12 أسبوع بواقع 1جم / کجم من وزن الجسم، وإعطاء مستخلص نبات الزنجبیل  بالفم مرة واحدة یومیاً لمدة 12 أسبوع بواقع 0.5 جم / کجم من وزن الجسم . وفی نهایة التجربة تم سحب عینات الدم لقیاس الإنسولین والجلوکوز والهیموجلوبین السکری، وأیضا دلائل الدهون والکولستیرول.

النتائج: أظهرت النتائج فی مجموعة البوال السکری إنخفاضاً ملحوظاً فی الإنسولین بالدم وارتفاع فی نسبة السکر والهیموجلوبین السکری، ووجود خلل فی دلائل الدهون بارتفاع مستوى الدهون الثلاثیة والکولستیرول والکولیسترول ذو الکثافة المنخفضة، وإنخفاض بمستوی الکولستیرول ذو الکثافة المرتفعة مقارنة بالمجموعة الضابطة. وباستخدام المعالجة بنباتی الحلبة والزنجبیل لوحظ تحسناً کبیراً ذو دلالة إحصائیة  فی معظم هذه الدلالات والقیاسات البیوکیمیائیة ورجوعها إلى المعدلات الطبیعیة دون حدوث فارقاً ذوأهمیة إحصائیة بین المجموعتین المعالجتین.

الاستنتاج: إستخدام نباتی الحلبة والزنجبیل  یثبط إرتفاع السکر بالدم والهیموجلوبین السکری وتخفیض حالات الشذوذ الأیضی فى حالات مرض البوال السکری.

 

REFERENCES
1. Abedinzade M, Nikokar I, Nasri S, Omidi JM and Nursabaghi F. (2013):  Effect of hexanic and alcoholic extract of fenugreek seed in male diabetic rats. Zahedan J. Res. Med. Sci. (ZJRMS), 15: 50-53.
2. Akhani SP, Vishwakarma Sl and Goyal Rk. (2005): Antidiabetic activity of zingiber officinale roscoe in streptozotocin-induced non-insulin dependent diabetic rats. Indain J. Pharm. Sci., 67 (5): 553-557.
3.  Amandeep S, Jaswant R and Devinder SM. (2016): Comparative evaluation of glipizide and fenugreek (Trigonella foenumgraecum) seeds as monotherapy and combination therapy on glycaemic control and lipid profile in patients with type 2 diabetes mellitus. International Journal of Basic & Clinical Pharmacology, 5 (3): 942-450.
4. Ansari R and Ansari S. (2011): Effectiveness of fenugreek for lowering the hemoglobin (HbA1c) in the patients with self-management of type 2 diabetes: A randomized control trial. Medical complication of type-2 diabetes. Dr. Collen Croniger (Ed). Pbl. InTech Europe: England: pp. 339-412.
5. Bunn HF. (1981): Evaluation of glycosylated haemoglobin diabetic patients. Diabetes, 30 (7):613-617.
6. Burstein RF and Scholnick VS. (1972):  Biochemistry and methodology of lipids. J Lipid Res., 25: 375-382.
7. Elshater AEA, Muhammad MAS and Mahrous MA M. (2009): Effect of ginger extract consumption on levels of blood glucose, lipid profile and kidney functions in Alloxan induced-diabetic rats. Egypt. Acad. J. Biology. Sci., 2: 153-162.
8.  Fauci AS, Braunwald E, Kasper DL, Stephen LH, Dan LL, Jameson JL and Loscalzo J. (2012): Harrison’s Principles of Internal Medicine.18th editors. pbl. New York: McGraw Hill: pp. 2968-3002.
9. Fossati P and Principe L. (1982): Serum triglycerides determined calorimetrically with an enzyme that produces hydrogen peroxide. Clin.Chem., 28: 2077-2080.
10. Friedewald WT, Levey RI and Fredrickson DS. (1972): Estimation of the concentration of low density lipoprotein cholesterol in plasma without use of the preparative ultracentrifuge. Clin. Chem., 18:499-502.
11. Giugliano D, Ceriello A and Esposito K. (2008): Glucose metabolism and hyperglycemia. Am J Clin Nutr., 87: 217-222.
12. Henry RJ, Cannon DC and Winkelman JW. (1997): Clinical Chemistry Principles and Tetchiness, pbl. New York: Harper and Row: p.1440.
13. Jafri SA, Abass S and Qasim M. (2011): Hypoglycemic effect of ginger (zingiber officinale) in alloxan induced diabetic rats (Rattusn norvagicus). Pak. Vet. J., 31: 160-162.
14. Kalaiselvi A, Reddy GA and Ramalingam V. (2015): Ameliorating Effect of Ginger Extract (Zingiber officinale Roscoe) on Liver Marker Enzymes, Lipid Profile in Aluminum chloride Induced Male Rats. IJPSDR, 7: 52-58.
15. Kazeem MI, Akanji MA, Yakubu MT and Ashafa AO. (2015): Antiglycation and Hypolipidemic Effects of Polyphenols from Zingiber officinale Roscoe (Zingiberaceae) in Streptozotocin-Induced Diabetic Rats. Tropical J. of Pharmaceutical Res., 14: 55-61.
16. Lebda MA, Taha NM,  Korshom MA, Mandour AA and El-Morshedy AM.( 2012): Biochemical effect of ginger on some blood and liver parameters in male Newzeland rabbits. Online J. Animal and Feed Research, 2: 197- 202.
17. Manmeet K, Narinder S, Geeta S and Davinder S. (2016): To study the efficacy and tolerability of fenugreek seed powder as add-on therapy with metformin in patients of type-2 diabetes mellitus. International Journal of Basic & Clinical Pharmacology, 5 (2): 378-383.
18. Marzouk M, Soliman AM and Omar TM. (2013): Hypoglycemic and antioxidative effects of fenugreek and termis seeds powder in streptozotocin-diabetic rats. European Review for Medical and Pharmacological Sciences, 17: 559-565.
19. Pandit R, Phadke A and Jagtap A. (2010): Antidiabetic effect of Ficus religiosa extract in streptozotocin induced diabetic rats. J Ethnopharmacol., 128: 462-466.
20. Reddy RR and Srinivasan K. (2011): Effect of dietary fenugreek seeds on biliary proteins that influence nucleation of cholesterol crystals in bile. ELSEVIER Steroids, 76: 455-463.
21. Shrivastava R, Solanki SS, Tomar V, Garud N, Garud A, Kannojia P and Jain N. (2009):  Comparative Evaluation of Polyherbal Combination for Hypolipidemic Activity. International Journal of Pharmaceutical Sciences and Drug Research, 17: 9-12.
22. Szkudelski T. (2001):  The mechanism of Alloxan and alloxan Action in β cells of the rats pancreas. Physiol Res., 50: 536- 546.
23. Trinder P. (1969): Determination of glucose in blood using glucose oxidase with an alternative oxygen acceptor. Ann. Clin. Biochem., 6: 24-33.