Effect of Equisetum arvense L. (Equisetaceae) in Microalbuminuria and Creatinine Excretion in Streptozotocin-Induced Diabetes in Male Rats
Shojaei Sadee Habib
In the present research, the methanolic extract of Equisetum arvense was analysed for its antidiabetic activity in streptozotocin-induced diabetic rats. The efficacy of the extract was also evaluated for protection of renal defects in diabetic rats. The blood glucose lowering activity of the methanolic extract was determined in steptozotocin-induced (50 mg kg-1, i.p.; dissolved in normal saline) diabetic rats, after oral administration in doses of 50, 100, 250 and 500 mg kg-1 daily for 5 weeks. Urine samples were collected before the induction of diabetes and at the end of 5 weeks of treatments and analyzed for urinary microalbumin and creatinine level. The data was compare statistically using one-way ANOVA tukey test. The results showed that in different doses of methanolic extract blood sugar decreased significantly in comparison with the treatment and control groups of diabetic rats. Also the weights of methanolic-extract treatment group were higher than the other treatment groups. The present studies clearly indicate a significant antidiabetic and renoprotective effect with the methanolic extract of Equisetum arvense and lend support for its traditional usage.
to cite this article:
Soleimani Safiyeh, Fathiazarbaijani Fathallah, Nejati Vahid, Shojaei Sadee Habib and Nanghshbandi Nabat, 2007. Effect of Equisetum arvense L. (Equisetaceae) in Microalbuminuria and Creatinine Excretion in Streptozotocin-Induced Diabetes in Male Rats. International Journal of Pharmacology, 3: 155-159.
It is well known that diabetes mellitus is the commonest endocrine disorder
that, according to the World than 176 million people world wide, in Mexico the
WHO estimates that the Health Organization (WHO, 2004), affects more number
of diabetic patients will increase from more than 2 million in 2002 to more
than 6 million in 2030, which would imply that in a few decades Mexico may have
highest rate of diabetes in the world. Because of the complications linked to
diabetes like heart disease, retinopathy, kidney disease and neuropathy, it
also is a common cause of chronic morbidity and disability among the working
population. The term diabetes mellitus describes a metabolic disorder of multiple
a etiologies and is characterized by chronic hyperglycaemia with disturbances
of carbohydrate, fat and protein metabolism resulting from defects in insulin
secretion, insulin action, or both. The causes of type 2 diabetes are either
insulin resistance with relative insulin deficiency or predominantly an insulin
secretory defect with or with out insulin resistance (WHO, 1999). Associated
changes in kidney function in terms of increased levels of blood urea, nitrogen
and distinct proteinuria have been reported in streptozotocin induced diabetic
rats (Pallavi et al., 2003). Detection of microalbuminuria or other
evidence of the microvascular implications of diabetes mellitus (DM) is unusual
before the onset of puberty, even though structural changes are occurring (Pascale
et al., 2004; Masateru et al., 2005). STZ-induced diabetes in
rats had been shown to be associated with functional and or morphological changes
in the kidney and liver (Urmila et al., 2003). After the introduction
of insulin therapy the field of herbal medicines research has been gaining significant
importance in the last few decades and the demand to use natural products in
the treatment of than 400 plant species showing anti diabetic activity, although
some of these many remain to be scientifically established (Nalamolu et al.,
2006). In north western regions of Iran Equisetum arvense used anti-diabetic
herbal remedy. Equisetum is a Asia and European herb which grows in moist
waste places throughout temperate regions of the world and is cultivated in
Iran. It is a member of a very primitive family of plants. In spring a spore-bearing
stem, resembling a thin asparagus shoot, rises 15-20 cm once shed, this is replaced
by a pale green bush with erect hollow jointed stems with longitudinal furrows
and with sharply-toothed sheaths covering each joint; from the sheaths of the
central stem arise whorls of fine branches, each giving off finer whorls, the
whole sometimes extending up to 60 cm in height, but usually less. Thiaminase
activity in Equisetum arvense extracts has been reported by Fabre et
al. (1993). Urinary metabolites of flavonoids and hydroxycinnamic acids
in humans after application of a crude extract from Equisetum arvense
reported by Graefe and Veit (1999). Enhancement of the division of E. arvense
protoplasts in culture by activated charcoal and their further development (Akira
et al., 1990) and the hepatoprotective effect of E. arvense has
been reported by Katikova et al. (2002). Hepatoprotective and free radical
scavenging activities of phenolic petrosins and flavonoids isolated from E.
Arvense (L) extract reported by Oh et al. (2004). Antidiabetic effect
of Equisetum arvense in diabetic rats reported by Soleimani et al.
(2007). The aim this study was to investigate the hypoglycemic effects of methanolic
extract in streptoztocin (STZ)-induced rats. We have found that administration
of E. arvense to streptozitocin-induced diabetic rats for month lowers
the level of serum glucose, urinary creatinine and microalbuminuria.
MATERIALS AND METHODS
Equisetum arvense L. subsp. arvense (Equisetaceae) pharmacopeial Name Equiseti herbars a traditional plant. Traditional use of the plant was recorded at market in 2005; by our selves of the plant for the treatment of kidney problems and diabetes in the Ardabil. Our own ethnopharmacological studies were performed in the community in Urmia University in 2005. Diabetic people were identified by the local health services and local healers. All informations were obtained about the plant and its special usage based on structured and unstructured interviews with the traditional healers and the diabetic people, respectively. The data were referred to plant samples (mini-herbarium) collected at its natural habitats and stored as herbarium vouchers for exact identification.
Plant material: Equisetum arvense is mainly found in northwestern regions of Iran especially Ardabil. This plant is considered an herbal drug and is used for renal disorders and diabetic related illness. This powdered from of this plant was purchased from the Herbal medicine research institute of Tabriz University, Iran and their identity was confirmed and voucher specimens were deposited at this Herbarium in Tabriz (No. 313).
Extraction: The dried plant was than milled to fine powder and 500 g of this powder was packed in to Soxhlet apparatus and extracted with methanol. The extract were dried at 45°C in hot air oven till solid to semisolid mass was obtained and were stored in airtight containers in refrigerator below 10°C.
Animals and experimental protocols: A total of 42 male wistar rats were used; eight weeks old weighing 150 to 200 g was obtained from the laboratory animal center of Veterinary faculty of Urmia University. The rats were housed under controlled environmental condition (12 12 hour's dark light cycle at 22°C) and had free access to standard rat chow and water. Diabetes was induced by intravenously injecting STZ (50 mg kg-1 body weight) in acetate normal saline. Two days later, induction of diabetes was con firmed by measuring the blood glucose level. One day prior to sacrifice, the rats were placed in metabolic cages to determine 12 h urinary microalbumin and creatinine excretion.
Experimental groups: The diabetic animals were classified into 6 groups (1-6) each of them with 6 rats.
Group 1 as non-diabetics control received 0.03 mL physiological NaCl-solution
Group 2 as the diabetic control received also 0.03 mL of physiological NaCl-solution
Group 3 were given the standard oral hypoglycemic agent glibenclamide (5 mg
kg-1 bw.) in the same vehicle
Groups (4-5-6 and 7) received (50, 100, 250 and 500 mg kg-1 bw.)
methanolic extracts, respectively.
Collection of blood and determination of blood glucose: Blood samples were taken from the tail vein before oral administration of the extracts or the vehicle. The glucose concentration was measured in plasma serum with Reflotron equipment and confirmed by Accutrend GC and Accu-check compact equipments (Roche).
Renal function: Urinary creatinine was measured by the jaffepicric acid procedure using Darman Kave kit absorbance at 490 nm was measured before and after addition of the acid reagent. In these and other clinical assays samples were diluted so that measurements were within the range of the standard curve. A portion of carefully timed (12 h collection period from 6:00 p.m. until 6:00 a.m., patients had avoided any physical activity) well-mixed sample of urine was used. Sample were stored at ±2 to ±8°C prior to testing. Analysis was made the same day after urine collection by immunoturbidimetric assay for urinary microalbumin Zist Chimiy on autoanalyser.
Statistical analysis: All the values of body weight, blood sugar and biochemical estimations were expressed as mean±standard error of mean (SEM) and analyzed for ANOVA tukey test.
The result of present field study confirmed that Equisetum arvense is
used as a hypoglycemic agent against diabetes type 1 and is also used against
kidney problems. The plant locally named by its Persian name Dome asb. In general,
the people drink the infusion of the plant after boiling 20-29 g in 1 L water.
The tea or the macerated in drunk during the day. The dry plant is sold at the
main market place in Urmia and Azarbaijans other place, with the same preparation
way and a similar recommended dose, further-more an methanolic preparation was
found, this preparation is recommended for the treatment of type 1 diabetes.
Those results confirm the previously reported use of E. arvense for the
treatment of diabetes type 1. STZ administration at of dosage of 50 mg kg-1
bw. to normal rats significantly elevated the blood glucose levels compared
with rats injected normal salin alone (Table 1) as in previous
reports (Andrae-Cetto et al., 2000; Andrae-Cetto and Wiedenfeld, 2001).
In present diabetic rats, the extracts showed significant hypoglycemic effects
(Table 1). The methanolic extract at doses of 50, 250 and
500 mg kg-1 bw. significant reduction (p<0.0001) of plasma glucose
level compared with diabetic control from the 1 and 5 weeks of treatment. The
methanolic extract at a dose of 100 mg kg-1 bw. did not show significant
activity of plasma glucose level compared with diabetic control. Gilbenclamide
(5 mg kg-1 bw.) produced a significant decrease in plasma glucose
(Table 1). These results indicate that there is no significant
difference between the tested plant preparations in comparison to glibenclamide
(standard hypoglycemic drug).
The mean weight of diabetic animals was significantly lower than that of non-diabetic animals (Table 1) how ever; the glibenclamide regimen was sufficient both to support weight gain in the diabetic animals during the early stages and to maintain their weight at the later stages of the study. Mortality was less than 6% among diabetic animals during the study the weights of the extracts-treated diabetic animals were statistically identical (Table 1).
The body weight of 5 weeks diabetic rats treated with methanolic extract of E. arvense at a dose of 100 mg kg-1 bw. treated not different from that of diabetic control rats. At the same time diabetics rats treated with methanolic extracts (50, 250 and 500 mg kg-1 day) immediately after diagnosis of diabetes had their body weight comparable to no-diabetic control groups (Table 1).
STZ-diabetic rats exhibited significantly lower urinary creatinine level compared to their respective non-diabetic control group (Table 1). Urinary creatinine level of diabetic groups treated with doses 50, 250 and 500 mg kg-1 bw. extract and gilbenclamide (5 mg kg-1 bw.) were significantly (p<0.001) higher compared to their respective diabetic control group. The urinary creatinine level in diabetic rats treated with methanolic extract of E. arvense at a dose of 100 mg kg-1 bw. treated not different from that of diabetic control rats. Table 1 show data for urinary microalbumin levels for healthy and diabetic rats. urinary microalbumin levels were significantly (p<0.05) increased in diabetic control. The urinary microalbumin levels in diabetic group treared with doses 50, 250 and 500 mg kg-1 bw. extract and diabetic group treated with 5 mg kg-1 bw. gilbenclamide were significantly (p<0.05) lower compared to their respective diabetic control group.
||Effect of oral administration of methanolic extract of
Equisetum arvense on plasma glucose concentration, body weight, urinary
creatinine and microalbumin in diabetic rats
|I-non diabetic control; II-diabetic control; III- diabetic
treated with 5mg kg-1 bw. gilbenclamide; IV-diabetic treated
with 50 mg kg-1 bw.; V-diabetic treated with 100 mg kg-1
bw.;VI- diabetic treated with 250 mg kg-1 bw.; VII- diabetic
treated with 500 mg kg-1 bw. extract. Values are given mean±SEM
for groups of six animals each. ap<0.0001, p<0.001,*p<0.05
(tukey-test), diabetic control was compared with the vehicle control and
extract treated groups were compared with the diabetic control
Diabetes mellitus is possibly the worlds largest growing metabolic
disorder and as the knowledge on the heterogeneity of this disorder is advanced,
the need for more appropriate therapy increases (Soleimani et al., 2007).
Hypoglycemic effect of Equisetum myriochaetum aerial parts has been reported
on streptozotocin diabetic rats (Andrae-Cetto et al., 2000). Similar
findings were reported by Revilla-Monsale el al. (2002) on type 2 diabetic
patients. In the present investigation, the methanolic extract of Equisetum
arvense was investigated for its antidiabetic and renoprotective activity
in streptozotocin-induced diabetic rats. Glibenclamide treatment (5 mg kg-1)
was not as effective in reducing blood glucose in STZ-diabetic rats as in normoglycaemic
rats. It has been reported that glibenclamide was not effective when destruction
of β-cells has occurred and hence more effective in moderate diabetic rats
than in severe diabetic animals (Andrade-Cetto et al., 2000; Hosseinzadeh
et al., 2002). The acute hypoglycaemic effect of glibenclamide results
has been shown from the stimulation of insulin release from the residual β-cells
and inhibition of glucagon secretion (Moller, 2001). The extract might possess
insulin like effect on peripheral tissues either by promoting glucose uptake
and metabolism or inhibiting hepatic gluconeogenesis. The phytochemical studies
of C. pentandra revealed the presence of epicatechin isolated from other
plants has been found to stimulate insulin secretion or possess an insulin-like
effect (Kameswara et al., 2001). In light of the results, present study
indicates that E. arvense have good antidiabetic activity. Methanolic
extract of E. arvense exhibited significant anti-hyperglycemic activities
in streptozotocin-induced hyperglycemic rats with out significant change in
body weight. They can also improve the condition of DB as indicated by parameters
like body weight. Among them methanolic extract produce a hypoglycemic effect
in rats. Similar finding reported by Soleimani et al. (2007). However
the improvement in urinary microalbumin and creatinine excretion with E.
arvense extract. Similar results were obtained with diabetic rabbits treated
with Eugenia jambolana (Kedar and chakrabarti, 1983) and non-diabetic
rats treated with Bauhinia forficate (Pepato et al., 2002). Renoprotective
effect of Terminalia chebula in diabetic rats reported by Nalamolu and
Srinivas in (2006). microalbuminuria and proteinuria typically reflect the presence
of moderate and advanced lesions, respectively in kidney disease, how ever,
the development of diabetic nephropathy is characterized by a progressive increase
in urinary protein particularly albumin and a late decline in glomerular filtration
rate, leading eventually to end-stage renal failure (Salah et al., 2004).
The pathophysiology involves glucose that binds irreversibly to proteins in
the kidney and circulation to form advanced glycosylation end products (AGEs).
AGEs can form complex crosslinks over years of hyperglycemia and can contribute
to renal damage by stimulation of growth and fibrotic factors via receptors
for AGEs. Increased glomerular capillary pressure occurs early in diabetes and
is associated with hyperfiltration at the glomerulus. The glomerular mesangium
expands, initially by cell proliferation and then by cell hypertrophy. Increased
mesangial stretch and pressure can stimulate this expansion, as can high glucose
levels. Mediators of proliferation and expansion include platelet-derived growth
factor and transforming growth factor β (TGF-β). TGF-β are particularly
important in the mediation of expansion and later fibrosis via the stimulation
of collagen and fibronectin. Angiotensi-II (AT-II) also contributes to the progression
of diabetic nephropathy. AT-II preferentially constricts the efferent arteriole
in the glomerulus, leading to higher glomerular capillary pressure. In addition
to its hemodynamic effects, AT-II also stimulates renal growth and fbrosis through
AT-II type1 receptors, which secondarily upregulate TGF-β and other growth
factors. The extract due to its significant hypoglycemic activity may have inhibited
the formation of advanced glycosylation end products. However the extract may
also have effect on the above stated other mechanisms. The possible mechanisms
behind the hypoglycemic activity and the inhibition of incidence of diabetic
nephropathy are yet to be studied.
In conclusion, the present studies indicated significant antidiabetic effects with the methanolic extract of Equisetum arvense (L.) and support its traditional usage in the control of diabetes and its complications. Further investigations to identify the active principle(s) are obviously needed together with a detailed evaluation on the mechanisms involved in the observed activities.
Akira, K., T. Masayuki, U. Saburo and M. Hisateru, 1990. Enhancement of the division of Equisetum arvense protoplasts in culture by activated charcoal and their further development. Plant Cell Physiol., 31: 999-1004.
Direct Link |
Andrade-Cetto, A. and H. Wiedenfeld, 2001. Hypoglycemic effect of Cecropia obtusifolia on streptozotocin diabetic rats. J. Ethnopharmacol., 78: 145-149.
Andrade-Cetto, A. and H. Wiedenfeld, 2004. Hypoglycemic effect of Acosmium panamense bark on streptozotocin diabetic rats. J. Ethnopharmacol., 90: 217-220.
Andrade-Cetto, A., H. Wiedenfeld, M.C. Revilla and I.A. Sergio, 2000. Hypoglycemic effect of Equisetum myriochaetum aerial parts on streptozotocin diabetic rats. J. Ethnopharmacol., 72: 129-133.
CrossRef | Direct Link |
Fabre, B., B. Geay and P. Beaufils, 1993. Thiaminase activity in Equisetum arvense and its extracts. Plant Med. Phytother., 26: 190-197.
Direct Link |
Graefe, E.U. and M. Veit, 1999. Urinary metabolites of flavonoids and hydroxycinnamic acids in humans after application of a crude extract from Equisetum arvense. Phytomedicine, 6: 239-246.
Direct Link |
Hosseinzadeh, H., M. Ramezani and A.R. Danaei, 2002. Antihyperglycaemic effect and acute toxicity of Securigera Securidaca L. seed extracts in mice. Phytoter. Res., 16: 745-747.
CrossRef | Direct Link |
Katikova, O.I., I. Kostin and V.S. Tishkin, 2002. Hepatoprotective effect of plant preparations. Eksp Klin Farmakol., 65: 41-43.
Direct Link |
Kedar, P. and C.H. Chakrabarti, 1983. Effects of jambolan seed treatment on blood sugar, lipids and urea in streptozotocin induced diabetes in rabbits. Indian J. Physiol. Pharmacol., 27: 135-140.
Masateru, I., M. Sonoo, A. Michitsune and H. Hiromichi, 2005. Nuclear factor-κB activation in diabetic rat kidney: evidence for involvement of P-selectin in diabetic nephropathy. Tohoku J. Exp. Med., 206: 163-171.
CrossRef | Direct Link |
Moller, D.E., 2001. New drug targets for type 2 diabetes and the metabolic syndrome. Nature, 414: 821-827.
CrossRef | Direct Link |
Nalamolu, K.R. and N. Srinivas, 2006. Anti-diabetic and renoprotective effects of the chloroform extract of Terminaria chebula Retz. seeds in STZ-induced diabetic rats. Biomed. J. BMC. Com., 14: 222-231.
Direct Link |
Oh, H., D.H. Kim, J.H. Cho and Y.C. Kim, 2004. Hepatoprotective and free radical scavenging activities of phenolic petrosins and flavonoids isolated from Equisetum arvense. J. Ethnopharmacol., 95: 421-424.
CrossRef | Direct Link |
Pallavi, V.L., N. Raghuram and S. Sivakami, 2003. Oxidative stress and genes expression of antioxidant enzymes in the renal cortex of streptozotocin-induced diabetic rats. Mol. Cellular Biochemist., 243: 147-152.
Direct Link |
Pascal, H.L., J. Sun, K. Davish and J. William, 2004. Langer-association of renal TGF-B and hypertrophy in female rats with diabetes mellitus. AJP. Renal Physiol., 287: F1011-F1020.
Pepato, M.T., E.H. Keller, A.M. Baviera, I.C. Ketellhut, R.C. Vendramini and I.L. Brunetti, 2002. Anti-diabetic activity of Bauhinia forficata decoction in streptozotocin-diabetic rats. J. Ethanopharmacol., 81: 191-197.
Rao, B.K., R Giri, M.M. Kesavulu and C. Apparao, 2001. Effect of oral administration of bark extracts of Pterocarpus santalinus L. on blood glucose level in experimental animals. J. Ethnopharmacol., 74: 69-74.
Revilla, M.C., A. Andrade-Cetto and S. Islas, 2002. Hypoglycemic effect of Equisetum Myriochaetum aerial parts on type 2 diabetic patients. J. Ethnopharmacol., 81: 117-120.
Salah, R., P. Pajica and M. Zliko, 2004. Microalbuminuria and Diabetes mellitus. Diabetologia Croat., 33: 209-220.
Shinde, U.A. and R.K. Goyal, 2003. Effect of chromium picolinate on histopathological alterations in STZ and neonatal STZ diabetic rats. J. Cell. Mol. Med., 7: 322-329.
Soleimani, S., F. Fathiazar Baijani, V. Nejati, H. Nazemiyeh and H.S. Sadee, 2007. Antidiabetic effect of Equisetum arvense L (Equisetaceae) in streptozotocin-induced diabetic in male rats. Pak. J. Biol. Sci., 10: 1661-1666.
WHO., 1999. Definition, diagnosis and classification of diabetes mellitus and its complications: Report of WHO consultation, Part 1: Diagnosis and classification of diabetes mellitus. WHO/NCD/NCS/99.2, World Health Organization, Geneva, Switzerland, pp: 1-59.