EVALUATION OF ANTIDIABETIC EFFICACY OF BROWN ALGA SPATOGLOSSUM ASPERUM J. AGARDH BY ALLOXAN STIMULATED HYPERGLYCEMIA ON WISTAR ALBINO RATSHTML Full Text
EVALUATION OF ANTIDIABETIC EFFICACY OF BROWN ALGA SPATOGLOSSUM ASPERUM J. AGARDH BY ALLOXAN STIMULATED HYPERGLYCEMIA ON WISTAR ALBINO RATS
S. Murugesan 1, M. Pandithurai 1, N. Krithika 2 and V. Sivamurugan * 3
Division of Algal Biotechnology and Bionano Technology, Post Graduate and Research Department of Botany 1, Post Graduate and Research Department of Chemistry 3, Pachaiyappa’s College, Chennai - 600030, Tamil Nadu, India.
Post Graduate and Research Department of Plant Biology and Plant Biotechnology 2, Quaid-E-Millath Government College for Women, Chennai - 600002, Tamil Nadu, India.
ABSTRACT: The present investigation aims to evaluate the effect of methanol extract of Spatoglossum asperum for its anti-hyperglycemic activity against diabetics induced by alloxan in Wistar albino rats. Artificially diabetes was induced in albino rats by the administration through injection of a single dose of alloxan monohydrate (10 mg/kg, I.P). The 10 mg/kg methanol residue of S. asperum was administered as a single oral dose per day to diabetes-induced rats for 28 days. The outcome of algae residue feeding was estimated by various biochemical and hematological parameters such as RBC, WBC, Hb, and platelets and lipid profile. After feeding of algal methanol extract of S. asperum in diabetic rats for 28 days, the blood glucose had significantly decreased, whereas the liver glycogen level had increased. Histopathological examination of the pancreas was done. The methanol residues of the Spatoglossum asperum possess very effective anti-hyperglycemic activity on the diabetic rats as compared to glipizide.
Brown algae, Spatoglossum asperum, Alloxan monohydrate, Diabetes mellitus
INTRODUCTION: Diabetes mellitus is a common disease affecting the citizens of both developed and developing countries. India is one of the leading countries for the number of people with diabetes mellitus, and it is estimated that diabetes would affect approximately 57 million people by the year 2025 in India. The main reasons of morbidity and mortality in the diabetic patients are cardiovascular diseases as patients are 2 to 4 times more susceptible to have heart disease and 5 times more likely to have a stroke 1, 2.
The control and treatment of diabetes and its complications mainly depend on the chemical and biochemical agents. Recently, the medicinal values of various plant extracts have been studied by many scientists in the field of diabetic research, and they contain valuable medicinal properties in different parts of the plant, and the major merits of herbal medicines seem to be their efficacy, low incidence of side-effects and low cost 3, 4.
Algae are a great source of natural compounds which are widely known and consumed in Asian countries 5. Seaweeds are considered as a source of bioactive compounds as they can produce a great variety of secondary metabolites characterized by a broad spectrum of biological activities, especially anti-diabetic activities 6, 7, 8, 9, 10. In this study, we attempted the anti-hyperglycemic effect of methanol extract of brown alga Spatoglossum asperum J. Agardhon alloxan stimulated hyperglycemic Wistar albino rats.
MATERIALS AND METHODS:
Collection of Marine Alga Spatoglossum asperum: Fresh materials of Spatoglossum asperum were obtained from Mandapam, the South East coast of Tamil Nadu and the algae were identified by the standard manual 11. The voucher specimen (PCCACL08) was deposited as Herbarium in the Department of Botany, Pachaiyappa’s College and Chennai- 600030. The salt and sand stuck on the surface of the freshly collected samples were removed by using sterilized seawater.
Extraction of Marine Alga: The shade dried seaweeds were finely powdered using a blender and stirred in with methanol in Soxhlet apparatus. The extract was evaporated to dryness under vacuum and dried in a vacuum desiccator. The crude methanol algal extract was stored at room temperature for further analysis 12, 13.
Selection and Acclimatization of Animals: The Wistar strain of male albino rats weighing between 180-220 g were used for this study. The animals were housed in large spacious cages and fed with commercial pellets and access to water ad libitum. The animals were well acclimatized to the standard environmental condition of temperature (25 ± 2 °C) and humidity (55 ± 5%) and 12 h light/dark cycles throughout the experimental period 12, 13.
Acute Oral Toxicity Study: In order to assess the toxic effects and tolerance limit and to determine a safe dose an acute oral toxicity study was carried out as per the CPCSEA guidelines. The rats fasted for 3-4 h before administration of extracts. All procedures described were reviewed and approved by the Institute Animal Ethical Committee (IAEC) no: IAEC/KMCP/153/FT/ 9599/2013-2014.
Induction of Diabetes Mellitus: Diabetes mellitus was induced in male albino Wistar rats by a single intraperitoneal injection of a freshly prepared solution of alloxan monohydrate (10 mg/kg body weight) in physiological saline after overnight fasting for 12 h 12, 13. The development of hyperglycemia in rats was confirmed by plasma glucose estimation 72 h post alloxan injection. The rats with fasting plasma glucose level of 160-220 mg/dL were used for this experiment.
Experimental Procedure: In the experiment, a total of 24 rats (18 diabetic surviving rats and 6 normal rats), each group with 6 rats was used. Diabetes was induced in rats 3 days before starting the experiment. The rats were divided into 4 groups after the induction of alloxan diabetes 12, 13.
Treatment Protocol: The experimental rats were divided into 4 groups of six rats as follows:
Group I: Normal control rats, received 0.9% saline at a dose of 10 mL/kg/body weight/rat/day for 28 days.
Group II: Alloxan induced diabetic rats, received 10 mg/kg body weight/rat/day of alloxan monohydrate through Intraperitoneal injection for 28 days.
Group III: Diabetic rats treated with glipizide (5 mg/kg/body weight/rat/day) dissolved in aqueous solution orally for 28 days 14.
Group IV: Diabetic rats, treated with methanol extract residue of S. asperum (10 mg/kg/body weight/rat/day) dissolved in aqueous solution orally for 28 days.
Histo-Chemical Parameters: After 28 days of treatment, the blood glucose level and the bodyweight of the experimental rats were measured. Then blood was collected retro-orbitally under mild ether anesthesia using capillary tubes. Blood was collected from the eyes (venous pool) by a sino-ocular puncture in fresh vials containing EDTA as anticoagulant agents and plasma were separated in a T8 electric centrifuge at 3000 rpm for 4 min. Then the animals were sacrificed by euthanasia. Liver and pancreas were immediately dissected out, washed in ice-cold saline to remove the blood and liver was used for estimation of enzyme activity while pancreas was subjected to histopathological studies. Blood, as well as tissue samples, were collected for the assessment of plasma glucose, hepatic glucokinase, hexokinase, glucose 6-phosphate, and phosphofructokinase content along with glycogen content.
Biochemical Analysis: Plasma insulin was determined by the ELISA method using a Boehringer-Mannheim kit with an ES300 Boehringer analyzer (Mannheim, Germany) 14. Hemoglobin was estimated by the method of Drabkin and Austin (1932) 15. Glycosylated hemoglobin (HbA1C) was estimated by following the method of Hongayo (2011). Hepatic glucokinase and hexokinase activity was assayed by means of glucose 6 phosphate-dependent spectrophotometric method Hongayo (2011). The glucose-6-phosphatase activity was estimated by calorimetrically Hongayo (2011). Glycogen content was determined colorimetrically by the method of Hongayo 16.
Parameters: Blood samples were assessed for RBC, WBC, HB, and platelets with an auto-analyzer (MISPA-EXCEL, Japan).
Estimation of Lipid Profile: Total serum cholesterol in the blood was estimated by using reported method 17. Estimation of HDL-C and LDL-C [Zawistowski, 2009] was done based on the following principle:
Phosphotungstate / Mg2+ precipitates chylomicrons, LDL and VLDL fractions. High-density lipoprotein (HDL) fraction remains unaffected in the supernatant. Cholesterol content of HDL fraction was assayed using Autozyme cholesterol. The total phospholipids in the serum were estimated by the method of the Post and Sen (1967) 17. Determination of triglycerides was done based on the principle that glycerol released from hydrolysis of triglycerides by lipoprotein lipase is converted by glycerol kinase into glycerol - 3 - phosphate, which is oxidized by glycerol phosphate oxidase to dihydroxyacetone phosphate and hydrogen peroxide. In the presence of peroxidase, hydrogen peroxide oxidizes phenolic chromogen to a red colored compound.
Histopathological Examination of Pancreas: The pancreas tissue section was fixed in 4 g/L formaldehyde and embedded in paraffin. Paraffin section was then stained with hematoxylin-eosin 17. Each sample was observed at 400 X magnification and scored according to the injuries.
Statistical Analysis: All the experimental values in this investigation were expressed as mean ± SD. All pair-wise multiple comparison procedures were attempted by Student-Newman-Keuls procedure using One-way ANOVA followed by SPSS 17.0. P<0.01 was considered to be significant. Triplicate assays were performed for each set of test conditions.
RESULTS: The in-vivo anti-diabetic activity of the methanol extract of marine brown alga S. asperumon alloxan-induced diabetes showed significant results.
Effect of Methanol Extract of S. asperumon Body Weight of Normal and Experimental Animals: Table 1 illustrates the bodyweight of the Group I to IV. The mean body weight of diabetic rats is 198.60 ± 2.80g which had significantly decreased to 156.40 ± 4.20 g, as compared to normal animal body weight (231.30 ± 5.80 g). The positive control, i.e. drug (Glipizide 5 mg/kg body weight) treated animals showed a recovery from bodyweight loss.
Treatment with methanol extract of S. asperum at a dose of 10 mg/kg body weight, exhibited an excellent recovery to keep the bodyweight comparable to that of normal control animals (Group I).
TABLE 1: EFFICIENCY OF THE METHANOL EXTRACT RESIDUE OF S. ASPERUM ON BODY WEIGHT OF NORMAL AND EXPERIMENTAL ANIMALS
|Groups||Initial body weight (g)||Final body weight (g)|
|Normal control - (Group I)||227.50 ± 4.50||231.30 ± 5.80|
|Toxic control - Alloxan monohydrate (Group II)||198.60 ± 2.80||156.40 ± 4.20(*a)|
|Positive control - Diabetic + Glipizide (Group III)||213.50 ± 2.50||215.30 ± 4.40(*b)|
|Treatment control – Diabetic + Methanol extract of S. asperum (Group IV)||226.35 ± 3.95||231.56 ± 3.68(*b)|
Effect of Methanol Extract of S. asperum on Blood Glucose Levels: Before Alloxan administration, the basal level of plasma glucose of the rats were not significantly higher in the rats selected for the study. In contrast, non-diabetic control (Group I) remained steadily euglycemic throughout the study. Alloxan induced diabetic animals (Group II) recorded more than twofold higher glucose (230.10 ± 5.70 mg/dL) as compared to that of normal animals (92.00 ± 2.30 mg/dL). At the end of the experiment, the diabetic induced animals had 138.1% of glucose than normal animals. Both the group III and IV, levels of glucose were restored to normal Table 2.
The methanol extract of S. asperum treated group IV (10 mg/kg body weight), showed a significant anti-hyperglycemic (P<0.001) effect at the end of the 14th day; the decrease in blood sugar was maximum on completion of the 28th day. Similarly, in group III animals, it also restored the blood sugar level nearly to that of the normal range.
TABLE 2: EFFICIENCY OF 4 WEEKS TREATMENT WITH METHANOL EXTRACT RESIDUE OF S. ASPERUM ON GLUCOSE LEVELS IN ALLOXAN DIABETIC RATS
|Groups||0 Day||14th Day||28th Day|
|Normal control - (Group I)||88.20 ± 5.20||93.50 ± 4.80||92.00 ± 2.30|
|Toxic control - Alloxon monohydrate (Group II)||160.20 ± 5.20||188.30 ± 4.70(*a)||230.10 ± 5.70(*a)|
|Positive control - Diabetic+Glipizide (Group III)||184.50 ± 4.20||154.80 ± 3.80(*b)||140.50 ± 2.30(*b)|
|Treatment control - Diabetic+Methanol extract of S. asperum (Group IV)||194.36 ± 4.23||162.30 ± 3.75(*b)||153.85 ± 2.55(*b)|
Unit: mg, No. of animals in each group (n) = 6, Values are expressed as Mean ± SEM. Values were found out by using One Way ANOVA followed by Newman Keul’s multiple range tests. (*a) Values were significantly different from normal control (GI) at P<0.001. (*b) Values were significantly different from diabetic control (GII) at P<0.001.
Effect of Methanol Extract on Glycogen Content: Glycogen content of liver tissue estimated on the 28th day in all the four groups of experimental animals Table 3 showed that in Group II, the liver glycogen content decreased significantly by 33.60% as compared to non-diabetic control animals. Whereas in group III and IV the liver glycogen level increased by 21.90 and 21.06% respectively.
TABLE 3: EFFICIENCY OF ADMINISTRATION OF METHANOL EXTRACT RESIDUE OF S. ASPERUM ON GLYCOGEN CONTENT OF LIVER TISSUE OF RATS
|Groups||Liver tissue glycogen content (mg/g)|
|Normal control - (Group I)||48.20 ± 2.80|
|Toxic control -
Alloxan monohydrate (Group II))
|14.60 ± 2.50(*a)|
|Positive control -
Diabetic + Glipizide (Group III)
|36.50 ± 5.20(*b)|
|Treatment control - Diabetic + extract of S. asperum (Group IV)||35.66 ± 1.34(*b)|
Effect of Methanol Extract on Hepatic Enzymes: To establish the diabetic condition, plasma glucose and glycogen content were determined 72 h after alloxan administration and the hepatic enzymes hexokinase, glucokinase and glucose-6-phosphate were estimated in all the groups (I to IV), 28 days after induction of diabetes Table 4. Non-diabetic control animals recorded notable values for enzyme hexokinase (0.252 ± 0.01), glucose-6-phosphate (0.143 ± 0.00) and glucokinase (30.50 ± 0.60).
Diabetic induced animals showed reduced levels of the enzymes hexokinase, glucose-6-phosphate, and glucokinase in their liver tissue as 0.100 ± 0.08, 0.146 ± 0.01 and 6.24 ± 0.11 respectively when compared to non-diabetic control. In group III and IV, the values are increased with respect to all the parameters. The observed levels between these two groups of animals were comparable with each other Table 4. However, they could not be restored to the levels observed in non-diabetic controls.
TABLE 4: EFFICIENCY OF ADMINISTRATION OF THE METHANOL EXTRACT RESIDUE OF S. ASPERUM ON HEPATIC ENZYMES INVOLVED IN CARBOHYDRATE METABOLISM IN RATS
|Normal control – (Group I)||0.252 ± 0.01||0.143 ± 0.00||30.50 ± 0.60|
|Toxic control - Alloxon monohydrate (Group II)||0.100 ± 0.08(*a)||0.146 ± 0.01(*a)||6.24 ± 0.11(*a)|
|Positive control - Diabetic + Glipizide (Group III)||0.167 ± 0.00(*b)||0.330 ± 0.00(*b)||21.24 ± 0.43(*b)|
|Treatment control - Diabetic + Extract of S. asperum (Group IV)||0.153 ± 0.00(*b)||0.297 ± 0.00(*b)||17.30 ± 0.38(*b)|
Unit = one unit is as 50% inhibition of NBT/mg protein. Unit = μmoles of H2O2 utilized/min/mg protein. Unit = μg/100 mg tissue. N = 6 in each group, values are expressed as Mean ± SEM. Values were found out by using One Way ANOVA followed by Newman Keul’s multiple range tests. (*a) Values were significantly different from normal control (GI) at P<0.001. (*b) Values were significantly different from diabetic control (GII) at P<0.001. Values with different superscripts in the same row differ significantly at 5% level P<0.05.
Effect of Methanol Extract of S. asperum on Haematological Parameters: The effect of methanol extract residue on parameters in diabetic induced rats (Group II) are shown in Table 5. At the end of the study period, there was a significant reduction of white blood corpuscles (9.10 ± 0.40 103 µL), red blood corpuscles (5.20 ± 0.14 106 µL), platelet counts (315.40 ± 20.14 103 µL) and slightly an increase in the haemoglobin (13.24 ± 0.60 gm/dL) level of the diabetic induced rats, compared to that of non-diabetic animals (Group I). No significant differences were seen in the mean value of WBC, RBC, HB and platelet counts in the positive control (Group III) and in the treatment control (Group IV) as compared to the non-diabetic animals.
TABLE 5: EFFICIENCY OF ADMINISTRATION OF THE METHANOL EXTRACT RESIDUE OF S. ASPERUM ON HAEMATOLOGICAL PARAMETERS
|Normal control - (Group I)||9.50 ± 0.60||5.30 ± 0.40||10.52 ± 0.46||312.20 ± 35.00|
|Toxic control - Alloxon monohydrate (Group II)||9.10 ± 0.40||5.20 ± 0.14||13.24 ± 0.60||315.40 ± 20.14|
|Positive control – Diabetic + Glipizide (Group III)||8.20 ± 0.40||5.20 ± 0.20||14.02 ± 0.40||291.40 ± 20.00|
|Treatment control – Diabetic + Extract of S. asperum (Group IV)||9.05 ± 0.70||5.15 ± 0.32||11.78 ± 0.32||292.25 ± 26.35|
Unit: For WBC -103/µL, RBC-106/µL, HB-gm/dL, Platelet-103/mL
Effect of Methanol Extract of S. asperum on Lipid Profile: The lipid profile Table 6 showed a significant increase in total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein (LDL) and phospholipids level, with a decrease in HDL level in alloxan-induced diabetic rats as compared to normal rats. Treatment of normal and alloxan-induced diabetic rats with the methanol extract residue of S. asperum at a dose of 10 mg/kg weight for 28 days resulted in a marked decrease in total cholesterol (117.50 ± 3.33 mg/dL), triglycerides (97.22 ± 2.55 mg/dL), low density lipoprotein (24.40 ±1.75 mg/dL) and phospholipids level (155.25 ± 3.92 mg/dL) and increase in HDL (41.50 ± 1.61 mg/dL) levels as compared to alloxan-induced diabetic rats that were equal to the positive control (Group III).
TABLE 6: EFFICIENCY OF ADMINISTRATION OF THE METHANOL EXTRACT RESIDUE OF S. ASPERUM ON CHOLESTEROL, TRIGLYCERIDE, HDL-C, LDL-C AND PHOSPHOLIPIDS
|Groups||Total Cholesterol (mg/dL)||Triglyceride
|High density lipoprotein (HDL-C (mg/dL)||Low density lipoprotein (LDL (mg/dL)||Phospholipids (mg/dL)|
|Normal control - (Group I)||83.20 ± 2.53||86.20 ± 1.75||48.92 ± 2.70||15.20 ± 3.20||122.50 ± 2.10|
|Toxic control - Alloxon monohydrate (Group II)||229.50 ±
|203.70 ± 5.80(**a)|
|Positive control - Diabetic + Glipizide (Group III)||103.90 ±
|146.80 ± 4.50(**b)|
|Treatment control - Diabetic + Extract of S. asperum (Group IV)||117.50 ±
|155.25 ± 3.92(**b)|
Values are expressed as mean ± SEM. Values were compared by using analysis of variance (ANOVA) followed by Newman-Keul's multiple range tests. (** a) Values are significantly different from toxic control G II at P<0.01. (** b) Values are significantly different from toxic control G II at P<0.01.
Histopathological Study: Histopathological observation Fig. 1a showed the normal acini and normal cellular population in the islets of Langerhans in the pancreas of normal rats (Group I). Extensive damage of islets of Langerhans in the pancreas of diabetic rats (Group II) and their reduction was observed Fig. 1b. The presence of lymphocytic infiltrates in the islet of Langerhans is characteristic of certain types of autoimmune disease.
The diabetic mice treated with glipizide showed restoration of the normal cellular population size of Islets with hyperplasia (Group III) Fig. 1c. Most of the nuclei were darkly stained than the surrounding acinar cells. The partial restoration of normal cellular population and enlarged size of β-cells with hyperplasia (Group IV) were observed Fig. 1d.
DISCUSSION: The present study was carried out to assess the anti-hyperglycemic property of the methanol extract of S. asperum in the control of diabetes on Wistar albino rats. Experimentally induced diabetes was used to analyze the biochemical, hormonal and morphological parameters during severe insulin deficiency or even during death.
In the present study, diabetes mellitus was induced by alloxan monohydrate at a dose of 10 mg/kg body weight. A significant increase in the plasma insulin level was observed when diabetic rats were treated with the methanol extract of S. asperum at a dose of 10 mg/kg body weight. In the case of uncontrolled diabetes rats, there is an increased number of proteins including hemoglobin and α-crystalline of the lens 18. Glycosylated hemoglobin (HbA1C) was found to increase in patients with diabetes mellitus to approximately 16%. 16 During diabetes, the excess glucose present in the blood reacts with hemoglobin. Therefore, the total level decreases in alloxan-induced diabetic rats 14, 16. Administration of the methanol extract of S. asperum at a dose of 10 mg/kg bodyweight for 28 days prevents a significant elevation in glycosylated, thereby increasing the level of total in diabetic rats. This could be due to the result of improved glycemic control produced by the extract of S. asperum. In the diabetes mellitus affected rats, proteins are used as an energy source; this leads to a decrease in protein storage, which in turn reduces the bodyweight 16. The decrease in body weight was observed in alloxan-induced diabetic rats 14. The administration of methanol extract of S. asperum at a concentration of 10 mg/kg body weight, led to an increase in the body weight in diabetic rats.
FIG. 1: PHOTOMICROGRAPH OF A CROSS-SECTION OF PANCREAS STAINED WITHHAEMATOXYLIN AND EOSIN (MAGNIFICATION × 400X). a) Normal Control (Group I); b) Toxic Control (Alloxan monohydrate) (Group II); c) Positive Control (Alloxan monohydrate + Glipizide) (Group III); d) Treatment group (Alloxan monohydrate + extract of S. asperum) (Group IV)
In the alloxan-induced diabetes mellitus rats, the rise in blood glucose was accompanied by an increase in serum cholesterol and triglycerides. The levels of cholesterol, triglycerides, and low-density lipoprotein (LDL) levels seemed to be nearly normal when treated with the methanol extract of S. asperum at a dose of 10 mg/kg body weight in the diabetic rats. Normalization of increased blood glucose by S. asperum extract suggests that it may enhance glucose transport across the cell membranes and stimulate glycogen synthesis or enhance glycolysis. Similarly, anti-hyperglycemic effects have been observed by the restoration of blood glucose level to normal in diabetic rats through different solvent extracts of brown alga Cystoseira moniliformis, 16 Padina arborescens 19 and Lobophora variegata 13.
There was an improvement in S. asperum extract and glipizide treated rats on liver protein levels, which might be due to the marked changes in circulating amino acid level, hepatic amino acid uptake and muscle output of amino acid concentrations 19. The increased hepatic glucose output in diabetes may be derived from glucogenesis 20, 21. The effect of methanol extract of S. asperum at a dose of 10 mg/kg on diabetic hypertriglyceridemia could be through its control of hyperglycemia. The improved glycemic control following sulfonylurea therapy decreases the levels of serum VLDL and total triglycerides 22.
The reduction in serum total cholesterol levels and dietary cholesterol absorption in the treated groups (fed with the algal extracts and drug) relative to the hypercholesterolemic control group was a likely explanation for the observed reduction in LDL-C levels 22. The elevated level of LDL-C got significantly reduced in the algal treated rats, which may be due to the antioxidant property of the extracts, which are capable of inhibiting the LDL-C peroxidation 23. The HDL-C is a free radical scavenger and prevents peroxidation of beta lipoproteins 24. Rats treated with algal extracts showed improved high levels of HDL-C, which may be due to the ability of the extract to accelerate the decomposition of free radical species generated during cholesterol administration 25, 26. The liver enzymes are normally found in circulation in small amounts because of the hepatic growth and repair. ALT, AST and ALP activities were elevated in hypercholesterolemic groups, meanwhile, the treated groups (IV) showed an obvious enhancement. The antihypocholesterolemic activity was due to the reduced cholesterol absorption in the gut 17.
The result of the present study clearly showed that the level of HDL-C was increased in alloxan-induced diabetic rats when treated with methanol extract residue of S. asperum at a dose of 10 mg/kg. These results confirm the reduction of intestinal glucose transport in-vivo and may be due to increased insulin sensitivity as observed in previous studies 27. Hexokinase is the prime enzyme catalyzing glucose phosphorylation 28. In the present study, hexokinase activity was found to be decreased in diabetic rats which may be due to insulin deficiency. Treatment with S. asperum elevated the activity of the hexokinase in liver and kidney. The S. asperum may stimulate insulin secretion, which in turn activates hexokinase, thereby increasing the utilization of glucose leading to decreased blood sugar levels. Seaweeds contain numerous bioactive substances that have been shown to lower cholesterol and blood pressure and promote healthy digestion and antioxidant activity 15. The presence of the bioactive metabolites in the marine brown alga S. asperum may explain the normalization of the FBS (Fasting blood sugar) level in diabetic rats. Dietary fucoxanthin also significantly reduced hepatic liver lipids and decrease in the lipid substrates for oxidation could result in the lower lipid hydroperoxide levels of the liver 8. The biochemical and parameters of Wistar albino rats after feeding with the methanol extract of S. asperum clearly indicated that it had not brought about any toxic impact on the rats. This was further confirmed by the quantification of total RBC, WBC and hemoglobin contents.
CONCLUSION: In the present study, the methanol extract of S. asperum at a dose of (10 mg/kg body weight) exhibited significant anti-hyperglycemic activity in alloxan-induced diabetic rats (Group IV). The extract showed improvement in the parameters like body weight, liver glycogen content and carbohydrate metabolizing enzymes as well as regeneration of β-cells of the pancreas in diabetes treatment. However, further pharmacological and biochemical investigations would elucidate the mechanism of action. The level of liver marker enzymes was also increased. The restoration of transaminases to their normal levels after the treatment also indicated a revival of insulin secretion. Histopathological examination reinforces the healing of the pancreas, by S. asperum methanol extracts, as a possible mechanism of their anti-diabetic activity.
ACKNOWLEDGEMENT: This work was financially supported by the University Grants Commission (UGC) New Delhi in the form of the minor research project.
AUTHORS’ CONTRIBUTION: All authors have contributed to the completion of this research work.
CONFLICT OF INTEREST: Authors have none to declare.
- Husni A: Therapeutic potential of seaweed polysaccharides for diabetes mellitus, seaweed biomaterials, Sabyasachi Maiti. Intechopen 2018; 27-45. doi: 10.5772/intechopen.76570.
- Radhika D and Priya: Assessment of anti-diabetic activity of some selected seaweeds: European Journal of Biomedical and Pharmaceutical Sciences 2015; 2(6); 151-54.
- Bakri R: Diabetes Epidemic in Malaysia, Second National Health and Morbidity Survey -Diabetes. Diabetes mellitus among adults aged 30 years and above. Public Health Institute 2007; 9: 7.
- Aguilera FJA, Ramos RR, Gutierrez SP, Contreras AA, Weber CCC and Saenz CCC: Study of the anti-hyperglycemic effect of plants used as anti-diabetic: J Ethno Pharma 2008; 61: 101-10.
- Lordan S, Ross RP and Stanton C: Marine bioactivities as functional food ingredients, potential to reduce the incidence of chronic diseases. Mar Drugs 2011; 9: 1056-00.
- Chew Y, Lim Y, Omar M and Khoo KS: Antioxidant activity of three edible seaweeds from two areas in South East Asia.LWT – Food Sci Technol 2008; 41: 1067-72.
- Thennarasan S, Murugesan S, Chidambaranathan N and Sivamurugan V: Analgesic, anti-inflammatory and anti-pyretic activity of the methanol extracts of brown alga variegata (J.V. Lamouroux) Womersley ex E.C. Oliveir. American J Phytomed Clinical Ther 2016; 4(2): 042-057.
- Woo MN, Jeon SM and Kim HJ: Fucoxanthin supplementation improves plasma and hepatic lipid metabolism and blood glucose Concentration in high-fat fed C57BL/6N mice. Chem Bio Inter 2010; 186: 316-22.
- Koneri R, Jha DK and Mubasheera MG: An investigation on the type I anti-diabetic activity of methanolic extract of marine algae, Gracilaria edulis & Sargassum polycystum. Int J Pharm Sci & Res 2018; 9(7): 2952-59.
- Rajeshkumar S, Divya, B and Malarkodi, C: Anti-diabetic activity of novel algal formulations ( tetrastromatica and Turbinaria conoides) against streptozotocin-induced diabetes in Albino rats. International Journal of Pharmaceutical Research 2017; 9(2): 54-58.
- Krishnamurthy V and Baluswami M: Phaeophyceae of India and neighborhood, Krishnamurthy institute of Algalogy, Chennai 2010; 1: 193.
- Murugesan S, Bhuvaneswari S and Sivamurugan V: In-vivo anti-diabetic efficacy of red algae Portieria hornemannii and Spyridia fusiformis methanol extract on alloxan stimulated hyperglycemic activity in Wistar albino rats. Saudi J Med Pharm Sci 2016; 2(10): 282-90.
- Thennarasan S, Murugesan S and Sivamurugan V: Anti-diabetic efficacy of methanol extract brown alga variegata on alloxan stimulated hyperglycemic Wistar albino rats. Asian J Pharm Tech 2017; 7(3): 157-65.
- Akbarzadeh S, Gholampour H, Farzadinia P, Daneshi A, Ramavandi B, Moazzeni A, Keshavarz M and Bargahi A: Anti-diabetic effects of Sargassum oligocystum on a streptozotocin-induced diabetic rat. Iranian Journal of Basic Medical Sciences 2018; 21: 342-46.
- Mohapatra L, Bhattamisra SK, Panigrahy RC and Parida SK: Evaluation of the antioxidant, hypoglycemic and anti-diabetic activities of some seaweed collected from the East Coast of India. Biomed & Pharmacol J 2016; 9(1): 365-75.
- Hongayo MC: The effect of brown alga Cystoseira moniliformis (KüTzing) Hauck extract on the blood glucose level of alloxan-induced hyperglycemic albino mice (Mus musculus Linne, 1758). J Phy Chem Sol 2011; 1: 1-17.
- Zawistowski J, Kopec A and Kitts DD: Effects of a black rice extract (Oryza sativa indica) on cholesterol levels and plasma lipid parameters in Wistar Kyoto rats. J Funct Foods 2009; 1(1): 50-56.
- Ponda MP: Moderate kidney disease inhibits atherosclerosis regression. Atherosclerosis 2010; 210(1): 57-62.
- Park MH and Han JS: Hypoglycemic Effect of Padina arborescens extract in streptozotocin-induced diabetic mice. Prevent Nutri Food Sci 2012; 17: 239-44.
- Klusonova P, Lenka P, Peter E, Ivan M, Josef Z, Jaroslav K and Jiri P: Local metabolism of glucocorticoids in Prague hereditary hypertriglyceridemic rats - effect of hypertriglyceridemia and gender. Steroids 2011; 76(12): 1252-59.
- Devaki T, Sathivel A and Raghavendran HRB: Stabilization of mitochondrial and microsomal function by polysaccharide of Ulva lactuca on D-Galactosamine induced hepatitis in rats. Chem Bio Interact 2009; 177(2): 83-88.
- Li-Qin T, Wei W, Li-Ming C and Sheng L: Effects of berberine on diabetes induced by alloxan and a high-fat/high-cholesterol diet in rats. J Ethno Pharmacol 2006; 108: 109-15.
- Yang C, Chen Y, Chen M, Jia R, Liu B and Zhao C: The anti-diabetic activity of brown seaweed Sargassum confusum polysaccharide hydrolysates in insulin resistance HepG2 cells in-vitro. Research Journal of Biotechnology 2017; 12(8): 1-10.
- Mohapatra L, Bhattamishra SK, Panigrahy RC, Parida S and Pati P: Anti-diabetic effect of Sargassum wightii and Ulva fasciata in high fat diet and multi low dose streptozotocin induced type 2 diabetic mice. UK Journal of Pharmaceutical and Biosciences 2016; 4(2): 13-23.
- Tam J: Peripheral CB1 cannabinoid receptor blockade improves cardiometabolic risk in mouse models of obesity. J Clin Invest 2010; 120(8): 2953-96.
- Sellamuthu PS, Muniappan BP, Perumal SM and Kandasamy M: Anti-hyperglycemic effect of mangiferin in streptozotocin-induced diabetic rats. J Health Sci 2009; 55: 206-14.
- Chanda M, Rajkumar V and Debidas G: Antidiabetogenic effects of separate and composite extract of seed of Jamun (Eugenia jambolana) and root of Kadali (Musa paradisiaca) in the streptozotocin-induced diabetic male albino rat. A comparative study. International J Pharmacol 2006; 2: 492-03.
- Akpan HB, Adefule AK, Fakoya FA and Caxton Martins EA: Evaluation of LDH and G6- PDH activities in auditory relay centers of streptozotocin-induced diabetic Wistar rats. J Analyt Sci 2007; 1: 21-25.
How to cite this article:
Murugesan S, Pandithurai M, Krithika N and Sivamurugan V: Evaluation of antidiabetic efficacy of brown alga Spatoglossum asperum J. Agardh by alloxan stimulated hyperglycemia on Wistar albino rats. Int J Pharm Sci & Res 2019; 10(9): 4322-29. doi: 10.13040/IJPSR. 0975-8232.10(9).4322-29.
All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
S. Murugesan, M. Pandithurai, N. Krithika and V. Sivamurugan *
Post Graduate and Research Department of Chemistry, Pachaiyappa’s College, Chennai, Tamil Nadu, India.
31 December 2018
18 April 2019
13 August 2019
01 September 2019