Abstract
Aims
Diabetes mellitus is a major manifestation of metabolic disorder which presents with hyperglycemia (high levels of serum blood sugar). In the present study, we aimed to investigate the effects of celery seed extract on different biochemical factors and histopathological changes in normal and streptozotocin (STZ)-induced diabetic rats.
Methods
A total of 35 male Wistar rats were divided into five groups (one normal and four diabetic groups). STZ was injected intraperitoneally to induce diabetes. The effects of hexane extract of celery seed and glibenclamide (as a positive control) were compared. Blood samples were analyzed on days 0, 18, and 33, and histopathological evaluations were performed at the end of the study.
Results
Glucose, triglycerides, and cholesterol levels significantly decreased, whereas insulin and high-density lipoprotein (HDL) levels increased in the extract-administered groups, as compared to the negative diabetic control group (P < 0.0001). The concentrations of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum of the extract-administered groups were significantly less than the negative control group (P < 0.0001). Histopathological reports revealed significantly less atrophy, necrosis, and inflammation in the rats receiving celery seed extract compared to the negative control group.
Conclusions
The results indicated that celery seed extract can be effective in controlling hyperglycemia and hyperlipidemia in diabetic rats, and demonstrated its protective effects against pancreatic toxicity resulting from STZ-induction.
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Introduction
Diabetes is a major manifestation of endocrine and metabolism disorders and hyperglycemia. Hyperglycemia and methylglyoxal can be implicated in development of micro- and macrovascular complications, which act as a silent killer worldwide [1]. Secondary diseases and long-term complications such as nephropathy, neuropathy, retinopathy and diabetic foot syndrome frequently occur in some diabetic patients [2]. Some factors such as serum uric acid independently predict the development of vascular complications in diabetic patients. Bjornstad et al. indicated that addition of serum uric acid to other risk factors (HbA1c, blood pressure and LDL) would improve the vascular event prediction over 6 years in adults with type I diabetes [3]. Diabetes type I affects pancreatic cells, which are responsible for producing insulin, by inhibiting immune response. However, in type II diabetes, cells become resistant to the use of insulin. Although pathogenesis of diabetes type I and type II is different, defects in insulin secretion and/or action, carbohydrates/lipids and proteins metabolism impairment are some of the main diabetic complications [4]. The number of diabetic patients is on a growing trend, and it is one of the major causes of mortality in most industrial countries [5]. While it was demonstrated that, in a 9-year period study, the regular follow-up diabetes mellitus patients had significantly lower glycemic burden and lower incidence of nephropathy and retinopathy [6], the World Health Organization (WHO) reported that 2.8 % of the global population sustained diabetes in 2000, and they estimated the number of diabetic patients would rise to 4.4 % of the global population by 2030 [7]. Furthermore, 6 % of Iranians aged 20–79 years are suffering from diabetes [8]. Diabetes is recognized as a major cause of cerebrovascular accident, cardiovascular problems, renal failure, atherothrombosis, myocardial infarction, blindness, and non-traumatic lower-limb amputations among the adult population [9]. Increased level of lipids in diabetes may cause cardiovascular complications [9]. Simultaneous decrease in antioxidant agents and mechanisms enhances oxidative stress, produces more free radicals and thereby causes diabetic complications [10–12].
In most cases, insulin and oral hypoglycemic drugs are administered as the first-line treatment in diabetic patients [10]. Due to the severe side effects of pharmacologic agents and absence of definitive treatments, researchers are trying to find traditional herbal extracts that have lower toxicity than chemical drugs [11]. On the other hand, for a very long time, medicinal plants were used for treatment of numerous diseases such as diabetes, icterus, cardiovascular problems, hypertension and cancer in traditional ways, especially in Eastern countries. A multitude of studies have been done to investigate the effects of various factors on diabetes management [12, 13]; however, medical knowledge packages for herbal medicines are not efficient enough. According to the previous surveys conducted in the USA and Australia, due to various reasons such as fewer side effects and suitable anti-diabetic properties, almost 50 % (in USA) and around 33 % (in Australia) of population prefer using herbal medicines [14]. Moreover, numerous plants such as garlic, crataegus, cayenne pepper and celery were administered for diabetes in traditional medicine [15–18]. Among these plants, celery (Apium graveolens) species, from the family of Apiaceae, has some synergistic beneficial effects on diabetes and hypertension [19], although no toxicologically significant effects have been reported for celery seeds. Moreover, unlike many dietary supplements, the available data suggest that celery seed extract does not significantly affect the enzyme systems and thus is less likely to alter the metabolism of drugs the patient may be taking [20, 21].
Celery leaves are odd-pinnate with dentate leaflets of 3–6 cm long and 2–4 cm wide on a central stem [22, 23]. Among the many constituents of celery, n-butylphthalide (NBP) along with sedanolide provides its aroma and taste, respectively [24]. Some researchers demonstrated that NBP, extracted from other medicinal plants, had some anti-hypertensive effects on animal models [25, 26]. Regarding former investigations, the most active compounds in celery were found in its seeds rather than the other parts (i.e., the roots and leaves) of the plant [27]. Other studies have demonstrated that celery seed and its active ingredients have different therapeutic properties such as hepatoprotective activity [28], cognitive enhancement [29] and antioxidant properties [30].
In our previous study, we exhibited that hexane extract of celery seed has significantly higher level of NBP compared to methanolic and aqueous ethanolic extracts [17]. In the present work, we investigated the anti-hyperglycemic effect of hexane extract of celery seed and its protective properties against pancreatic tissue damage in streptozotocin (STZ)-induced diabetic rats. Different variables including weight, daily water intake, fasting blood sugar (FBS), insulin, triglyceride, cholesterol, high-density lipoprotein (HDL), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were evaluated, and histopathology of pancreatic cells and islets was reported.
Materials and methods
Chemicals
The celery seeds were purchased from Imam Pharmacy, Mashhad, Iran, and their identity was confirmed by the herbarium of School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. Glibenclamide (Tehran Chemie Pharmaceutical Co., Iran), glucose diagnostic kit, insulin radioimmunoassay kit, triglyceride and cholesterol quantitation kits, HDL quantitation kit, ALT and AST activity assay kits (Pars Azmun Co., Iran), STZ (ENZO Life Sciences, USA), normal saline, ethanol (96 %), liquid paraffin, n-hexane (Merck, Germany), xylazine (Loughrea Corporation, Ireland), ketamine (Retexmedica GmbH, Germany), formalin, distilled water (Hakim Pharmaceutical Co., Iran) and pelleted feed (Javaneh Khorasan Co., Iran) were obtained.
Preparation of the celery seed extraction
Dry celery seeds (50 g) were powdered and suspended in n-hexane (250 ml) at room temperature. The solvent was shaken for 48 h in the dark. Then, it was passed through a cotton filter, the coarse particles were separated, and clear and green suspension remained. Thereafter, the suspension was centrifuged (2594g for about 10 min). The supernatant was separated and evaporated to dryness in the dark at room temperature. Finally, the green oil remained and kept in refrigerator [17].
Animals
Thirty-five adult male Wistar rats (220–270 g), aged 10–11 weeks, were obtained from the animal facilities of the Pharmaceutical Research Center, BuAli Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran. The animals were housed in cage with a 12-h light/12-h dark cycle at 21 ± 2 °C, and free access to food and water was provided for them. The experimental procedures for all the animals were in accordance with the Ethics Committee Acts of Mashhad University of Medical Sciences.
Induction of diabetes by STZ
STZ with chemical formula of 2-deoxy-2-(3-methyl-3 nitrosourea)-d-glucopyranose was used to induce diabetes in the rats. All the groups except group 1 (normal group) intraperitoneally received STZ (80 mg/kg) on day zero (72 h before extract injections). STZ solution was prepared in cold normal saline and was administered to overnight-fasting rats (12 h before injection). On day 3, only those animals with FBS above 200 mg/dl were selected as diabetic animals and were randomized into treatment and negative control groups. Blood samples for checking glucose level were taken from eye orbital plexus.
Experimental design
About 35 rats were randomized into five groups (n = 7) as follows: (1) normal group (not diabetic rats); (2) positive control group [diabetic rats receiving glibenclamide (1 mg/kg) intraperitoneally every other day for 1 month]; (3) negative diabetic control group (receiving 0.5 ml liquid paraffin, as an extract carrier, intraperitoneally every other day for 1 month); (4) and (5) diabetic animals (receiving 100 mg/kg and 200 mg/kg n-hexane extracts intraperitoneally every other day for 1 month, respectively). The animals were fasted from 12 h before blood sampling. Then, they were anesthetized by a mixture of ketamine (60 mg/kg) and xylazine (6 mg/kg), and blood samples were collected from their eye orbital plexus. The blood samples were evaluated on days 3 (the day before STZ injection), 18 and 33. The blood samples were exposed to room temperature for 20 min, centrifuged at 4000 rpm for 10 min and preserved at −20 °C until biochemical assay (biochemistry laboratory in Mashhad University of Medical Sciences) was performed. Different blood serum factors such as FBS, insulin, HDL, cholesterol, triglyceride, ALT and AST were evaluated.
Histopathology
On the final day (day 33), the rats were killed and their pancreas was removed and fixed in buffered formalin 10 % solution for conducting histopathological assay. After processing and embedding, the pancreases were cut into 5-µm slices. The slices were stained with hematoxylin and eosin as the routine staining method. Sliced tissues of each sample were studied at total magnification of 100× and 200×. Different changes in pancreas tissue such as atrophy, necrosis, fibrosis, insulitis and decrease in Langerhans islets were evaluated.
All the qualitative data were changed into quantitative information (Table 4). Scores 0–4 were given to different levels of tissue damage in histopathological evaluations as follows: score zero was considered for normal cells without any damage, and scores one, two, three and four were considered for mild (<25 %), moderate (<50 %), serious (<75 %) and severe (>75 %) damages, respectively.
Statistical analysis
Prism software was employed to analyze the data, and the mean ± SEM were reported. Two-way analysis of variance (ANOVA) and Bonferroni post hoc test were performed for biochemical parameters. The normality test was performed for pathological data. One-way ANOVA and Tukey’s post hoc test were applied for parametric data. Moreover, Kruskal–Wallis and Dunn’s post hoc tests were performed for nonparametric data. P value less than 0.05 was considered significant.
Results
Effects of celery seed extract on weight and daily water intake
Table 1 demonstrates body weight changes in all the groups during the study. Our data showed that body weight in the normal and treatment groups significantly increased after 1 month, whereas a slight increase in body weight was observed in the negative control group. The maximum increase in body weight was observed in the normal and celery extract-receiving groups (200 mg/kg). As shown in Table 2, the initial volume of water intake varied between 35 and 37 ml in all the groups, while the value significantly changed in different groups on day 33. The maximum increase in water intake (59 ml) was seen in the negative control group, whereas the minimum change was reported in the normal (5.5 ml) and positive control (7 ml) groups. The water intake change values (34 and 19.7 ml in groups 4 and 5, respectively) in the groups receiving celery seed extract were significantly different from that of the negative control group.
Effect of celery seed extract on FBS
Figure 1 exhibits FBS of the rats during the experiment. No difference was observed between the groups at the beginning of the study, whereas glucose level decreased on days 18 and 33 in all the treatment and positive control groups. The rate of FBS decrease in the positive control group and the groups receiving 200 and 100 mg/kg celery seed extract was 53, 57 and 38 %, respectively.
Effect of celery seed extract on insulin
Table 3 shows the comparison of the level of serum insulin in all the experimental groups on days 0 and 33. As can be noted, insulin concentration in the negative control group significantly decreased, while the normal group did not exhibit any remarkable alterations in insulin level during the experiment. As compared to the negative control group, glibenclamide and celery seed extract (100 and 200 mg/kg) significantly elevated serum insulin concentration in the diabetic rats (P < 0.001).
Effect of celery seed extract on triglyceride
As shown in Fig. 2, no significant difference was observed between the groups in terms of triglyceride serum level at the beginning of the study. Except for the normal group, a remarkable elevation in triglyceride concentration was noted in all the diabetic groups on day 18. From day 18 to day 33, a significant decrease in triglyceride level was observed in the groups receiving celery seed extract and the positive control group, whereas triglyceride concentration had continuously increased in the negative control group during the study (P < 0.001). On day 33, the minimum triglyceride level was observed in the positive control group, which received glibenclamide. No significant change was reported in the normal group during the study.
Effects of celery seed extract on cholesterol and HDL
Figures 3 and 4 illustrate the changes in serum cholesterol and HDL levels in different groups. Cholesterol level had gradually increased from 67.85 to 72 mg/dl in the negative control group, while this value decreased slowly from 69 to 61 mg/dl in the positive control group. In the groups receiving celery seed extract, cholesterol level increased from day 0 to day 18 and then decreased from day 18 to day 33.
On day 33, no significant difference was observed between the positive control group and the group receiving 200 mg/kg celery seed extract. HDL gradually decreased during the study (from 54 to 45 mg/dl) in the negative control group. However, from day 0 to day 33, HDL level elevated in the other groups. On day 33, no significant difference between the normal and treatment groups (positive control group and the groups receiving celery seed extract) was observed in terms of HDL level.
Effects of celery seed extract on ALT and AST
Figures 5 and 6 show the serum concentrations of ALT and AST in different groups. ALT and AST levels significantly increased after STZ-induced diabetes in the negative control group (P < 0.001). Administration of glibenclamide and celery seed extract prevented elevation of ALT and AST enzymes in the rat serum. The difference between the negative control group and the other groups was statistically significant (P < 0.001).
Histopathology
Histopathological evaluations were carried out to study the effects of celery seed extract on STZ-induced damages to pancreas tissue such as atrophy, necrosis, insulitis, fibrosis and reduction in the number of Langerhans islets (Fig. 7). All the images were presented with two magnifications (100× and 200×). Figure 7a, b demonstrates the normal pancreatic cells and Langerhans islets in the normal rats. As shown in Table 4 and Fig. 7a, b, minimum changes were observed in the normal group. In contrast, maximum damages and pathological scores were reported in the negative control group (Table 4). Figure 7c, d shows a significant decrease in the number of Langerhans islets, necrotic residues of the pancreatic cells and presence of inflammatory cells in the Langerhans islets. Glibenclamide and celery seed extract remarkably reduced the histopathological damages and scores, as compared to the negative control group (P < 0.05). Figure 7e, f illustrates mild inflammatory and degenerative changes in the Langerhans islets of the group receiving 200 mg/kg celery seed extract.
Discussion
Diabetes mellitus is a systemic, chronic disease defined as prolonged hyperglycemia and characterized by metabolic disorders of carbohydrates, proteins and lipids. A great number of people suffer from diabetes worldwide [31]. It seems that increasing the free radicals and oxidative stress involved in development of diabetes, whereas the protective agents and mechanisms such as antioxidants are impaired [32]. Methods that induce oxidative stress are usually applied to evaluate hypoglycemic effects of drugs in animal models [33]. In several studies, a single dose of STZ as a glucosamine–nitrosourea compound was used to induce diabetes mellitus in animals by destruction of the pancreatic islets. Intraperitoneal injection of an appropriate dose of STZ causes a reduction in insulin secretion from pancreas due to oxidative damage of Langerhans islets. In this model, insulin is not completely absent but is markedly depleted [34, 35].
Due to the unpredictable side effects and high rate of secondary failure of conventional anti-diabetic agents, the effects of herbal medicines have been evaluated to prevent and control diabetes mellitus [36]. In diabetes mellitus, water intake usually increases, while the rate of weight gain significantly diminishes [32, 37]. In this study, no significant difference was observed between the groups in terms of water intake on the initial day (day zero). After diabetes induction, as expected, water consumption dramatically increased in the diabetic negative control group; moreover, glibenclamide and celery seed extract could decrease water intake in the diabetic animals during the study (in comparison with the negative control). In comparison with the positive control group, the volume of water consumption in the groups receiving celery seed extract was still high on day 33 (P < 0.01), which can be due to the diuretic effect of the active components of celery seed extract. Other studies have also reported antispasmodic and diuretic effects of celery seed extract [17, 38, 39]. Tables 1 and 2 indicate that glibenclamide and celery seed extract could significantly enhance weight gain and decrease the volume of water intake in the diabetic rats compared to the negative control group (P < 0.05 and P < 0.001, respectively). In a study by Al-Sa’aidi et al. [40], body weight in the rats receiving n-butanol extract of celery seed considerably increased compared to diabetic negative control group. In STZ-induced diabetic rats, degradation and loss of structural proteins were the main reasons of body weight reduction [41]. Regarding the previous studies, degradation and loss of structural proteins might have a direct relationship with weight loss in diabetic patients; consequently, active compounds in celery seed may prevent loss of proteins. Other researchers have reported that liver weight decreases with diabetes due to enhanced catabolic processes such as glycogenolysis, lipolysis and proteolysis. The glycogen content in the liver is a storage of glucose and must be considered as an important indicator of diabetes [42]. It was reported that some other herbal extracts such as Nigella sativa might be effective in reducing hyperglycemia and increasing body weight due to its effects on reducing protein loss [43].
The present study showed that administration of celery seed extract from day three to the final day (day 33) significantly lowered serum glucose and prevented insulin depletion in the intervention groups, as compared to the negative control group (P < 0.0001). No significant difference was observed in glucose level of the glibenclamide (1 mg/kg)-administered and celery seed extract (100 and 200 mg/kg)-administered groups on day 33. Serum insulin in the treated rats increased significantly compared to the negative control group, which implies the protective effect of celery seed against oxidative stress and histopathological injuries in pancreas tissue after administration of STZ.
Previous investigations on diabetes mellitus reported that impaired oxidant–antioxidant balance causes oxidative stress, which results in molecular and cellular tissue damage. In other words, increased production of oxygen free radicals (due to non-enzymatic and autoxidative glycosylation) attack macromolecules (nucleic acids, lipids, proteins, and carbohydrates) and cause oxidation injury, which in turn changes oxidant–antioxidant balance in diabetic patients [44]. Moreover, previous studies have suggested a correspondence between the hypoglycemic effects of some medicinal herbs such as Salvia officinalis L., Bridelia ndellensis and Urtica dioica on blood glucose and their antioxidant properties [42, 45, 46]. In addition, antioxidant, cyclooxygenase and topoisomerase inhibitory compounds were found in celery seed by Momin et al. [30, 40]. Lin et al. [47] reported that apigenin and luteolin extracted from celery seed had inhibitory effect on the aldose reductase enzymes (with a key role in the polyol pathway). The polyol pathway appears to be involved in diabetic complications such as nephropathy, retinopathy and neuropathy [48]. Aldose reductase catalyzes the reduction of glucose to sorbitol, which cannot cross the cell membranes, and its accumulation in cells produces osmotic stresses by drawing water into the tissues. Unused glucose enters polyol pathway and reduces to sorbitol [42]. Therefore, apigenin and luteolin in celery seed can lower the microvascular complications of diabetes through inhibition of aldose reductase. Other studies demonstrated that some flavonoids such as apigenin and luteolin, which are present in N. sativa, U. dioica, Punica granatum and celery seed, have antioxidant and protective effects [32, 49–53]. Thus, anti-hyperglycemic effects of celery seed may be due to increased secretion of insulin, proliferation of β-cells or increasing their repair after STZ-induced damage, enhanced glucose transport into cells and its utilization by tissues, increased glycogen synthesis from glucose in the liver and improved oxidant–antioxidant balance [54]. Serum glucose level in group 5 (receiving 200 mg/kg celery seed extract) was comparable to that of the positive control group (receiving glibenclamide) on day 33, which indicated the efficacy of the hexane extract of celery seed in lowering blood glucose level.
Additionally, the level of triglycerides in the groups receiving celery seed extract increased from day 3 to day 18, whereas it decreased from day 18 to day 33. In diabetes management, reduction in triglyceride serum level takes a longer duration of time, as compared to serum glucose. This triglyceride reduction pattern was observed in the glibenclamide-administered rats as the positive control. In the healthy rats, lipoprotein lipase is activated by insulin, which hydrolyzes triglycerides; hypertriglyceridemia happens due to insulin deficiency after STZ injection in diabetic rats [55]. Thus, increased insulin concentration reduced deactivation of lipoprotein lipase in the rats receiving celery seed extract and consequently lowered the level of serum triglyceride on the final day [42]. Furthermore, in the previous studies, samples with insulin deficiency were unable to activate the lipoprotein lipase enzyme, which led to hyperglyceridemia [55].
The results showed that the higher dose of celery seed extract (200 mg/kg) significantly lowered the total cholesterol level, while HDL level increased after the administration of 100 and 200 mg/kg celery seed extract. Hypercholesterolemia and HDL reduction were also observed in the negative control group, which indicated dyslipidemia in the diabetic rats. These results imply that celery seed extract administration could effectively improve the metabolism of carbohydrates, lipids and proteins in diabetic patients; metabolic disorders are usually observed in diabetic patients, and hypercholesterolemia and hypertriglyceridemia were observed in the STZ-induced diabetic rats [36, 56, 57]. High level of total cholesterol and low level of HDL play an important role in the incidence of cardiovascular diseases [58]. The effect of celery seed extract was similar to that of glibenclamide. Hypolipidemic effect of celery seed and its bioactive compounds were demonstrated in a former study by Cheng et al. [59]. The hypolipidemic effects of ethanolic extract of celery seed were evaluated by Mansi et al., and their results were in agreement with ours [60]. In a study by Iyer et al., ethanolic extract of celery seed and its chloroform fraction reduced total cholesterol, low-density lipoprotein and triglyceride and increased HDL more significantly than its aqueous fraction in hyperlipidemic rats. Phytochemical evaluations of A. graveolens indicated the presence of tannin, terpenoid, alkaloid, flavonoid, glycosides and sterols, which may be responsible for its hypolipidemic activities [61].
Our results indicated that the rats receiving 100 and 200 mg/kg celery seed extract had stable levels of ALT and AST, whereas these values dramatically increased in the diabetic negative control group. Increased concentration of AST and ALT showed that diabetes may induce liver dysfunction. Quite in line with our results, Larcan et al. found that liver was necrotized in diabetic patients. Elevated ALT and AST concentrations in serum may be due to the leakage of these enzymes from hepatic tissue into the blood [62], which demonstrates the hepatotoxicity of STZ. Therefore, treatment of diabetic rats with celery seed extract could effectively reduce AST and ALT concentrations in rats. Other studies have also reported the protective effect of celery seed extract on liver function and its lowering effect on hepatic enzymes [28, 63]. In the present study, no adverse effect or toxicity was observed after administration of celery seed extract in diabetic rats.
According to our histopathological results, celery seed extract (especially in 200 mg/kg dose) could significantly decrease fibrosis, insulitis, necrosis and atrophy in pancreas tissue compared to the negative control group. It could also prevent attenuation of Langerhans islets (Table 4). These data revealed that active components of celery seed extract could diminish oxidative stress, which was induced by STZ in the diabetic rats. It may also induce regeneration of the islets in the pancreas tissue. Figure 7 shows that the diameter and number of islet cells dramatically increased in the extract-administered groups compared to the negative control group. From the histopathological point of view, treatment with 200 mg/kg celery seed extract could significantly increase regeneration of the pancreatic cells with negligible degeneration and necrosis in the remaining cells compared to the negative control and normal groups. Other studies demonstrated that celery had reasonable amounts of NBP, especially in seeds. Several protective effects of NBP have been reported in previous studies. In a study by Peng et al, inhibitory effects of NBP on lipid peroxidation were reported. Antioxidant activities of NBP may be involved in neuroprotection and treatment of Alzheimer’s disease [29]. NBP has been shown to have anti-inflammatory properties [64]. In an in vitro study, NBP was demonstrated to have protective effects against neurotoxicity induced by 1-methyl-4-phenylpyridinium (MPP+) in PC12 cells, which is a cellular model for Parkinson’s disease [65]. Previous works revealed anti-ischemic properties of NBP, which could reduce the risk of cerebrovascular accident and prevent oxidative stress and mitochondrial damages [66]. NBP could also suppress the release of cytochrome C, to induce the up-regulation of vascular endothelial growth factor and subsequently, decrease oxidative stress in diabetic rats [67]. Hepatoprotective effect of celery seed extract against paracetamol and thioacetamide intoxication in rats was previously reported [28]. Singh et al. concluded that stimulation of hepatic regeneration may be involved in hepatoprotective properties of celery seed extract. Other mechanisms such as activation of reticuloendothelial system and inhibition of biosynthesis of some proteins may explain its protective role in different tissue toxicities [28]. Other active components such as apigenin and lutein may explain the protective properties of celery seed extract against oxidative stress and different tissue toxicities [34, 41]. According to the present study and previous reports, antioxidant properties and regeneration stimulating and anti-inflammatory activities of some active components of celery seed extract (e.g., NBP, apigenin, and lutein) may justify its protective effects against STZ-induced pancreatic toxicity.
Conclusion
This study indicated that administration of celery seed extract could significantly reduce water intake and glucose, cholesterol and triglyceride levels in STZ-induced diabetic rats, while it increased serum insulin and HDL compared to the negative diabetic rats. It prevented ALT and AST elevation in the diabetic animals. Histopathological evaluations revealed protective activities of celery seed extract against atrophy, necrosis, insulitis, fibrosis and decrease in the number of Langerhans islets induced by STZ in the diabetic rats.
Different active ingredients such as NBP, apigenin and lutein (through several mechanisms such as antioxidant, anti-inflammatory and regeneration inducing effects) may contribute to protective properties of celery seed extract against STZ-induced pancreatic toxicity. Our results indicated anti-diabetic and anti-hyperlipidemic effects of celery seed extract, which requires further investigation, especially in human samples.
References
Jensen TM et al (2015) Impact of intensive treatment on serum methylglyoxal levels among individuals with screen-detected type 2 diabetes: the ADDITION-Denmark study. Acta Diabetol 52(5):929–936
Bergis D et al (2014) Coronary artery disease as an independent predictor of survival in patients with type 2 diabetes and Charcot neuro-osteoarthropathy. Acta Diabetol 51(6):1041–1048
Bjornstad P et al (2014) Serum uric acid predicts vascular complications in adults with type 1 diabetes: the coronary artery calcification in type 1 diabetes study. Acta Diabetol 51(5):783–791
Hull RL et al (2004) Islet amyloid: a critical entity in the pathogenesis of type 2 diabetes. J Clin Endocrinol Metab 89(8):3629–3643
Iyer A, Chan V, Brown L (2010) The DOCA-salt hypertensive rat as a model of cardiovascular oxidative and inflammatory stress. Curr Cardiol Rev 6(4):291–297
Anjana RM et al (2014) Regularity of follow-up, glycemic burden, and risk of microvascular complications in patients with type 2 diabetes: a 9-year follow-up study. Acta Diabetol 52(3):601–609
Wild S et al (2004) Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. Diabetes Care 27(5):1047–1053
Hasani-Ranjbar S, Larijani B (2014) Medicinal plants as potential new target drugs in endocrine disorders-review article. Iranian J Public Health 43(1):24–34
Heather LC, Clarke K (2011) Metabolism, hypoxia and the diabetic heart. J Mol Cell Cardiol 50(4):598–605
Kamal M, Jawaid T (2010) Pharmacological activities of Lawsonia inermis Linn.: a review. Int J Biomed Res 1(2):37–43
Palsamy P, Subramanian S (2009) Modulatory effects of resveratrol on attenuating the key enzymes activities of carbohydrate metabolism in streptozotocin–nicotinamide-induced diabetic rats. Chem Biol Interact 179(2):356–362
Larijani B (2007) The clinical investigation of Securigera securidaca (l.) Degen & doerfler seeds in treatment of type II diabetic patients: a randomized, double-blind, placebo-controlled study. Diabetes 56:A562
Mahmoodabadi NA et al (2006) Preventive effect of hydroalcoholic Cynara scolymus extract on appearance of type 1 diabetes mellitus in male rats. J Diabetes Metab Disord 6(1):175
Kim JD et al (2007) Ameliorative anti-diabetic activity of dangnyosoko, a Chinese herbal medicine, in diabetic rats. Biosci Biotechnol Biochem 71(6):1527–1534
Jurikova T et al (2012) Polyphenolic profile and biological activity of Chinese hawthorn (Crataegus pinnatifida BUNGE) fruits. Molecules 17(12):14490–14509
Soleymanifard S et al (2014) An investigation of the effects of raw garlic on radiation-induced bystander effects in MCF7 cells. Iranian J Med Phys 11(4):350–357
Moghadam MH, Imenshahidi M, Mohajeri SA (2013) Antihypertensive effect of celery seed on rat blood pressure in chronic administration. J Med Food 16(6):558–563
Renault S et al (2003) CAY-I, a novel antifungal compound from cayenne pepper. Med Mycol 41(1):75–82
Broadhurst CL, Polansky MM, Anderson RA (2000) Insulin-like biological activity of culinary and medicinal plant aqueous extracts in vitro. J Agric Food Chem 48(3):849–852
Powanda M, Rainsford K (2011) A toxicological investigation of a celery seed extract having anti-inflammatory activity. Inflammopharmacology 19(4):227–233
Powanda MC, Whitehouse MW, Rainsford KD (2015) Celery seed and related extracts with antiarthritic, antiulcer, and antimicrobial activities, in novel natural products: therapeutic effects in pain, arthritis and gastro-intestinal diseases. Springer, Berlin, pp 133–153
Domagała-Świątkiewicz I, Gąstoł M (2012) Comparative study on mineral content of organic and conventional carrot, celery and red beet juices. Acta Sci Pol Hortorum Cultus 11(2):173–183
Zhou K et al (2009) Triterpenoids and flavonoids from celery (Apium graveolens). J Nat Prod 72(9):1563–1567
Houston MC (2005) Nutraceuticals, vitamins, antioxidants, and minerals in the prevention and treatment of hypertension. Prog Cardiovasc Dis 47(6):396–449
Zhu J, Zhang Y, Yang C (2015) Protective effect of 3-n-butylphthalide against hypertensive nephropathy in spontaneously hypertensive rats. Mol Med Rep 11(2):1448–1454
Dimo T et al (2003) Possible mechanisms of action of the neutral extract from Bidens pilosa L. leaves on the cardiovascular system of anaesthetized rats. Phytother Res 17(10):1135–1139
Popović M et al (2006) Effect of celery (Apium graveolens) extracts on some biochemical parameters of oxidative stress in mice treated with carbon tetrachloride. Phytother Res 20(7):531–537
Singh A, Handa S (1995) Hepatoprotective activity of Apium graveolens and Hygrophila auriculata against paracetamol and thioacetamide intoxication in rats. J Ethnopharmacol 49(3):119–126
Peng Y et al (2012) L-3-n-butylphthalide reduces tau phosphorylation and improves cognitive deficits in AβPP/PS1-Alzheimer’s transgenic mice. J Alzheimers Dis 29(2):379–391
Momin RA, Nair MG (2002) Antioxidant, cyclooxygenase and topoisomerase inhibitory compounds from Apium graveolens Linn. seeds. Phytomedicine 9(4):312–318
Zimmet P, Alberti KG, Shaw J (2001) Global and societal implications of the diabetes epidemic. Nature 414(6865):782–787
Alimohammadi S et al (2013) Protective and antidiabetic effects of extract from Nigella sativa on blood glucose concentrations against streptozotocin (STZ)-induced diabetic in rats: an experimental study with histopathological evaluation. Diagn Pathol 8:137
Eddouks M, Chattopadhyay D, Zeggwagh NA (2012) Animal models as tools to investigate antidiabetic and anti-inflammatory plants. Evid Based Complement Altern Med 2012:142087. doi:10.1155/2012/142087
Battu G et al (2007) Hypoglycemic and anti-hyperglycemic effect of alcoholic extract of Benincasa hispida in normal and in alloxan induced diabetic rats. Pharmacogn Mag 3(10):101
Pari L, Saravanan G (2002) Antidiabetic effect of Cogent db, a herbal drug in alloxan-induced diabetes mellitus. Comp Biochem Physiol C: Toxicol Pharmacol 131(1):19–25
Pushparaj PN, Tan BKH, Tan CH (2001) The mechanism of hypoglycemic action of the semi-purified fractions of Averrhoa bilimbi in streptozotocin-diabetic rats. Life Sci 70(5):535–547
Chatterjea M, Shinde R (2012) Textbook of medical biochemistry. Jaypee Brothers Medical Publishers, New Delhi, India
Fazal SS, Singla RK (2012) Review on the pharmacognostical and pharmacological characterization of Apium graveolens Linn. Indo Glob J Pharm Sci 2(1):36–42
Abbaskhan A et al (2012) Biological activities of Indian Celery, Seseli diffusum (Roxb. ex Sm.) Sant. & Wagh. Phytother Res 26(5):783–786
Al-Sa’aidi JA, Alrodhan MN, Ismael AK (2012) Antioxidant activity of n-butanol extract of celery (Apium graveolens) seed in streptozotocin-induced diabetic male rats. Res Pharm Biotechnol 4(2):24–29
Veeramani C, Pushpavalli G, Pugalendi KV (2008) Antihyperglycaemic effect of Cardiospermum halicacabum Linn. leaf extract on STZ-induced diabetic rats. J Appl Biomed 6:19–26
Akram E, Maryam E (2009) Antidiabetic effects of sage (Salvia officinalis L.) leaves in normal and streptozotocin-induced diabetic rats. Diabetes Metab Syndr: Clin Res Rev 39:40–44
Kanter M et al (2004) Effects of Nigella sativa on oxidative stress and β-cell damage in streptozotocin-induced diabetic rats. Anat Rec A: Discov Mol Cell Evol Biol 279(1):685–691
Ramakrishna V, Jailkhani R (2007) Evaluation of oxidative stress in Insulin Dependent Diabetes Mellitus (IDDM) patients. Diagn Pathol 2(1):22–30
Sokeng S et al (2005) Antihyperglycemic effect of Bridelia ndellensis ethanol extract and fractions in streptozotocin-induced diabetic rats. Afr J Trad CAM 2(2):94–102
Das M et al (2011) Antihyperglycemic and antihyperlipidemic activity of Urtica dioica on type 2 diabetic model rats. J Diabetol 2(2):1–6
Lin L-Z, Lu S, Harnly JM (2007) Detection and quantification of glycosylated flavonoid malonates in celery, Chinese celery, and celery seed by LC-DAD-ESI/MS. J Agric Food Chem 55(4):1321–1326
Forbes JM, Coughlan MT, Cooper ME (2008) Oxidative stress as a major culprit in kidney disease in diabetes. Diabetes 57(6):1446–1454
Miura K, Kikuzaki H, Nakatani N (2002) Antioxidant activity of chemical components from sage (Salvia officinalis L.) and thyme (Thymus vulgaris L.) measured by the oil stability index method. J Agric Food Chem 50(7):1845–1851
Golalipour MJ et al (2011) Protective effect of Urtica dioica L. (Urticaceae) on morphometric and morphologic alterations of seminiferous tubules in STZ diabetic rats. Iranian J Basic Med Sci 14(5):472
Nencu I et al (2012) Preliminary research regarding the therapeutic uses of Urtica dioica L. Note I. The polyphenols evaluation. Farmacia 60(4):493–500
Lansky EP, Newman RA (2007) Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. J Ethnopharmacol 109(2):177–206
Jang S, Kelley KW, Johnson RW (2008) Luteolin reduces IL-6 production in microglia by inhibiting JNK phosphorylation and activation of AP-1. Proc Natl Acad Sci USA 105(21):7534–7539
Joseph B, Jini D (2011) Insight into the hypoglycaemic effect of traditional Indian herbs used in the treatment of diabetes. Res J Med Plant 5(4):352–376
Taskinen MR (1987) Lipoprotein lipase in diabetes. Diabetes/Metab Rev 3(2):551–570
Association AD (2008) Diagnosis and classification of diabetes mellitus. Diabetes Care 31(Supplement 1):S55–S60
Tan BK, Tan CH, Pushparaj PN (2005) Anti-diabetic activity of the semi-purified fractions of Averrhoa bilimbi in high fat diet fed-streptozotocin-induced diabetic rats. Life Sci 76(24):2827–2839
Winocour P et al (1986) Lipoprotein abnormalities in insulin-dependent diabetes mellitus. The Lancet 327(8491):1176–1178
Cheng M-C et al (2010) Chemical synthesis of 9 (Z)-octadecenamide and its hypolipidemic effect: a bioactive agent found in the essential oil of mountain celery seeds. J Agric Food Chem 58(3):1502–1508
Mansi K et al (2009) Hypolipidemic effects of seed extract of celery (Apium graveolens) in rats. Pharmacogn Mag 5(20):301
Iyer D, Patil U (2011) Effect of chloroform and aqueous basic fraction of ethanolic extract from Apium graveolens L. in experimentally-induced hyperlipidemia in rats. J Complement Integr Med 8(1). doi:10.2202/1553-3840.1529
Concepción NM et al (1993) Free radical scavenger and antihepatotoxic activity of Rosmarinus tomentosus. Planta Med 59(4):312–314
Sultana S et al (2005) Inhibitory effect of celery seeds extract on chemically induced hepatocarcinogenesis: modulation of cell proliferation, metabolism and altered hepatic foci development. Cancer Lett 221(1):11–20
Peng Y et al (2007) l-3-n-Butylphthalide improves cognitive impairment induced by chronic cerebral hypoperfusion in rats. J Pharmacol Exp Ther 321(3):902–910
Liu K et al (2012) Protection against neurotoxicity by an autophagic mechanism. Braz J Med Biol Res 45(5):401–407
Li L et al (2009) DL-3-n-butylphthalide protects endothelial cells against oxidative/nitrosative stress, mitochondrial damage and subsequent cell death after oxygen glucose deprivation in vitro. Brain Res 1290:91–101
Zhang T, Jia W, Sun X (2010) 3-n-Butylphthalide (NBP) reduces apoptosis and enhances vascular endothelial growth factor (VEGF) up-regulation in diabetic rats. Neurol Res 32(4):390–396
Acknowledgments
We gratefully thank the vice chancellor of research, Mashhad University of Medical Sciences, for financial and logistic support of this Project through Grant Number 910889. This result was obtained from a Pharm. D. thesis (Dr. Masoud Ramezani).
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Tashakori-Sabzevar, F., Ramezani, M., Hosseinzadeh, H. et al. Protective and hypoglycemic effects of celery seed on streptozotocin-induced diabetic rats: experimental and histopathological evaluation. Acta Diabetol 53, 609–619 (2016). https://doi.org/10.1007/s00592-016-0842-4
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DOI: https://doi.org/10.1007/s00592-016-0842-4