Journal of Rawalpindi Medical College (JRMC); 2013;17(1):134-137

Original Article Antihyperglycemic/ Hypoglycemic Effect of (Ajwain / Ajmod) in Streptozotocin induced Diabetic Rats

Khalid Niaz*, Seemi Gull**. M. Azam Zia*** *Department of Pharmacology and Therapeutics, Islamabad Medical and Dental College Islamabad; ** Department of Pharmacology and Therapeutics, Rawalpindi Medical College, Rawalpindi; *** Department of Pharmacology and Therapeutics, Azad Jammu & Kashmir Medical College, Muzaffarabad Azad Kashmir

Abstract In the last few years herbal medicines are under extensive research. Many traditional medicines in use To investigate the effect of Apium Background: now days are derived from medicinal , minerals graveolen L. (Celery seeds) on blood glucose and insulin ,4 concentrations in streptozotocin (STZ) induced diabetic and organic matter. The World Health Organization rats. (WHO) has listed 21,000 plants, which are used for medicinal purposes around the world.5,6 Celery (Apim Methods: Forty Sprague Dawely rats weighing from 180 to 250 grams were randomly divided into four graveolens) is a member of the Umbelliferous () experimental groupsA,B,C &D, each containing ten (10) Family and related to family of . Celery seeds are rats. Diabetes was induced by intraperitoneal commonly known as Ajwain krufs and in Urdu it is administration of STZ (60mg/kg) in groups B,C&D,while called as Ajmod. Celery is a highly consumed group A served as normal control. The experimental vegetable with a high flavonoid content. Studies of animals became diabetic within 48 to 72 hours after celery seeds have identified the flavonoids, 7-0-apiosyl- administration of STZ.,Group B rats were taken as diabetic glycosides and 7-0-glucosides of luteolin, apigenin and control. Alcoholic of Celery seeds, the test drug was administered (400mg/kg) orally to experimental Chryseoriol as phenolic acids, chologenic acid, 6,7 gruop,C,while group D received standard drug cinnamic acids, coumarins and their glycosides. glibenclamide.induced diabetic rats for six weeks. Normal Antihyperglycemic effects of Ajwain are studied by control groupA& diabetic control B received only normal different workers,but not in Pakistan. 8-19 The present saline solution orally. Blood samples were collected from investigation sets out to study the antihyperglycemic experimental groups after 43 days,24 hours of the last dose. activities of Celery seeds extract and other related ANOVA and Tukey HSD (Honestly Significant biochemical parameters in normal and streptozocin Difference) test applied to all groups. diabetic animals. The hypoglycemic effects of celery Results: The results show Celery seeds extract treatment seeds were compared with Glibenclamide a standard caused a statistically significant decrease in the elevated drug. serum glucose levels and increase in the serum insulin concentrations in test group C as compared to group B . Material and Methods Conclusion: Findings of present study provide Forty Sprague Dawely rats (twenty females and evidence for traditional use of apium. graveolen in the control of diabetes. twenty males) weighing from 180 to 250 grams were obtained from the National Institute of Health, Key Words: Streptozotocin , Apium graveolen . Islamabad. For acclimatization these animals were

kept for 7-15 days in the experimental research Introduction laboratory,University of Health Sciences Lahore, at a Diabetes Mellitus is a clinical disorder characterized controlled temperature of 20–25°C and 45 to 65% by increased blood glucose due to deficiency or humidity. The animals were divided into four groups decreased secretion of insulin. About 180 million of the (A, B, C, and D). Each group consisting of 10 rats each, world population have diabetes mellitus.2 In 1995, 4.3 in these (group A, and B) served as a control and the million people had DM and in 2025 it will increase to other two (C, and D) as experimental groups. 14.5 million. This data suggests that Pakistan becomes Group-A :Rats were fed on Purina rat chow and the fourth country among the top 10 countries of the normal saline (1.5ml) orally for six weeks of world.1-3 experimental study.

134 Journal of Rawalpindi Medical College (JRMC); 2013;17(1):134-137

Group B: In rats of this group diabetes mellitus was significantly less than group A. The body weights of induced by giving an intraperitoneal injection of STZ, group C (STZ induced Diabetes + Celery seeds extract) 60mg/kg, after an overnight fasting. No drug or were less than group A, but it was still significantly extract was given to this group. These rats were fed on greater than the STZ induced diabetic rats in group B. Purina rat chow and normal saline (1.5ml) orally for Similarly the body weights of group D (STZ induced six weeks of experimental study. Diabetes+ Glibenclamide) were also increased Group-C: In these diabetic rats celery seeds extract significantly(Table 1) was given 400mg/kg as a single dose orally for six In group C rats at the end of experimental study weeks of experimental study. mean blood glucose level SEM.was 139.65 0.257 Group-D: In these diabetic rats glibenclamide mg/dl in Group C rats compared with groups Group- (600µg/kg) body weight was given orally for six B (Diabetic group). Mean blood glucose values were weeks. statistically significant p 0.001, when fasting blood For the preparation of ethanol extract of celery glucose levels of Group C (STZ induced Diabetes + seeds, freshly collected were shade-dried and Celery seeds extract) was compared with groups coarsely powdered in a grinder. Powdered dried seeds Group-B (Diabetic group) and other groups(Table 2) 500g were soaked for 12hrs in 2.0 liters of ethyl alcohol. Then this suspension was filtered and the Table 1. Effect of Celery seeds extract on the residue was again soaked in equal amount of ethanol body weight of normal and STZ-diabetic rats. for 48hrs and again filtered. The two filtrates obtained (Mean SEM;n=10 rats from each group) were evaporated and dried by distillation under Groups 0th day 43rd day reduced pressure at 40 to 50°C in rotary evaporator. The ethanol extract so obtained (42g) was suspended 208 6.46 249.4 3.99 in distilled water in the required amount at the time of Group A (normal dosing. control) Initial blood sample for glucose estimation was Group B (STZ induced 205 6.83 176.9 1.82 collected from all groups A, B, C, and D, from the tail Diabetic control) veins of rats.Blood sample from group B, C, and D was Group C (STZ induced 200.5 1.74 233.3 6.46 collected after 72 hours of STZ-injection and diabetes Diabetes + celery seeds mellitus confirmed by Ames One Touch Glucometer. extract given) Animals with blood glucose more than 200mg/dl were Group D (STZ induced 205.5 6.16 230 6.30 selected for this research study. Diabetes + After 24hours of the last dose of celery seeds Glibenclamide given) extract and after overnight fasting (12 to 14 hours) the animals were anesthetized under chloroform vapors. Table 2. Comparison of all groups for plasma Blood samples (5ml) were collected by cardiac glucose (mg/dl). (Mean±SEM; n=10 for each group). puncture from all groups. Blood was collected in Mean Group Group B Group C Group D(STZ vacutainers and then centrifuged for ten minutes at a Plasma A (STZ (STZ induced speed of 5000 rev/min. Plasma was separated and Glucos (normal induced induced Diabetes + o e control) Diabetic Diabetes Glibenclamid stored at -20 C and used for the measurement plasma (mg/dl) control) + celery e given) glucose and plasma insulin. The significance between seeds two means was calculated by the ANOVA and Tukey extract HSD. The difference was regarded statistically given) significant if the ‘p’ value was < 0.05. 0th day 78.88- 80.0+6.8 77.7+1.7 83.5+4.50 +1.5 Results 43rd day 78.88+1. 254.7+1.8 139.6+1.6 At the end of experimental study, body weights of 5 2 5 102.8+1.82 rats in group A (normal control) were increased as compared to their initial body weights.Significant There was statistically significant difference( p body weight loss was observed in the STZ induced 0.001) in mean fasting plasma insulin levels of Group diabetic rats in group B (STZ induced Diabetic control C (STZ induced Diabetes + Celery seeds extract) group), when their final body weights were compared compared with groups Group B (Diabetic group). with their initial body weights. The final body weights Group C was also compared with groups (Group A, of group B (STZ induced Diabetic control group) were and D). The mean fasting plasma insulin levels of

135 Journal of Rawalpindi Medical College (JRMC); 2013;17(1):134-137

Group C (STZ induced Diabetes + Celery seeds extract) reported as the active principles.7 Apigenin, isolated could not reach the level as that of the normal control from celery seeds extract also possess an inhibitory effect group A, but it justifies the role of this extract in on the aldose reductase enzymes.22 This enzyme is induction of insulin release from pancreatic islets( known to play a key role in the polyol pathway, by Table 3) catalyzing the reduction of the glucose to sorbitol, which cannot diffuse out of cell membranes .In diabetics Table 3. Comparison of all Groups for plasma excessive intracellular accumulation of sorbitol result in Insulin ( L/ml) insulin (µL/ml) ±SEM of all group (n=10 for each group). chronic complications such as neuropathy, retinopathy, Mean Group B Group C Group and cataract. A study was conducted in which Plasma Group A (STZ (STZ D(STZ antihyperglycaemic effects of herbal drugs Insulin (normal induced induced induced Cardiospermum halicacabum L were investigated in ( L/ml) control) Diabetic Diabetes Diabetes 19 control) + celery + STZ induced diabetic rats. This plant has same active seeds Glibencla compounds apigenin and luteolin,10 and has same effect extract mide on insulin release from pancreatic islets of rats as that of given) given) celery seeds.7

0th day 11.68+2.0 11.31+1.5 10.81+2.5 11.04+1.8 Scientific studies have been conducted in which antihyperglycaemic effects of herbal drug L was reported. Aegle marmelos L. investigated in STZ rd 43 day 11.53+2.0 4.422+2.0 8.913+4.0 10.93+3.0 induced diabetic rats showed results are similar to the

results of our current study. 22,23 In celery seeds treated

diabetic rats, the significant elevation of plasma insulin Discussion may be due to the stimulation of insulin secretion from Diabetes is a chronic metabolic disorder affecting a the existing β-cells of the pancreas.20 The decrease in major population worldwide. A sustained reduction in blood glucose level of diabetic rats treated with celery hyperglycemia will decrease the risk of developing seeds extract might be due to elevated secretion of microvascular complications. The conventional insulin, increasing repair or proliferation of β-cells, therapies for diabetes have many shortcomings like enhancing the effect of insulin and increasing the unwanted side effects and high rate of secondary failure. oxidative capability which in turn, increases the On the other hand, herbal are expected to have utilization of glucose by the tissues.24 Extrapancreatic similar efficacy with minimum side effects like than mechanisms such as enhanced glucose transport into the those of conventional drugs. 20,21 cells and improved formation of glycogen in the liver Streptozotocin is a glucoseamine-nitrosourea might be involved in celery seeds induced decrease in compound used in medicinal research to produce animal blood glucose concentration of STZ diabetic rats.24 models for diabetes mellitus. As with other alkylating Several of the compounds found in celery seeds have agents in the nitrosourea class, it is toxic to cell by been reported as possessing antidiabetic activity, like causing damage to the DNA, though other mechanisms flavonoids, such as luteolin and apigenin, which not may also contribute. Thus STZ injection results in only have well documented antidiabetic activity but also diabetes mellitus due to the destruction of beta cells of have significant role in the prevention of diabetes islets of Langerhans.20 Streptozocin selectively destroys mellitus complications.25,26 the pancreatic insulin secreting β-cells, leaving less active cells and resulting in a diabetic state. STZ-induced References diabetes is characterized by severe loss in body weight, 1. United Nations .Diabetes. New York; World Health and this reduction is due to loss or degradation of Organization: 2006. structural proteins, as the structural proteins are known 2. Shera AS, Jawad F, Maqsood A. Prevalence of diabetes in Pakistan. Diabetes Research and Clinical Practice 2007; 76: to contribute to body weight. This effect is evident by 219-22. high level of glucose in animals. 20 Flavonoids are one of 3. Rajkumar L, Ggovindarajulu P. Increased degradation of the most numerous and widespread groups of phenolic dermal collagen in diabetic rats. Idian J Exp Biol 1991; 29; compounds in higher plants. Some of these, due to their 1081-83. 4. Modak M, Dixit P, Londhe J, Ghaskadbi S, Devasgayam TPA. phenolic structure, are known to be involved in the Indian and herbal drugs used for the treatment of healing process of free radical mediated diseases diabetes. J. Clin. Biochem. Nutr 2007; 40: 163-73. including diabetes. Celery seeds possess several 5. Grover, JK, Yadav S, and Vats V. Medicinal plants of with antidiabetic potential. J. Ethnopharmacol 2002; 81: 81- flavonoids such as apigenin, and luteolin, which are 100.

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