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Original article / research
Year : 2023 Month : January-March Volume : 12 Issue : 1 Page : BO06 - BO12

Effect of Rosuvastatin and Glibenclamide in Alone or in Combination on Glucose Homeostasis in Diabetic Male Albino Wistar Rats: An Observer-blinded Experimental Interventional Study


Suganya Ganesan, Balaji More, Selvalaxmi Gnansegaran, Isswarya Anandan, Nitya Selvaraj, Jayaram Mohanasundaram
1. Assistant Professor, Department of Pharmacology, Vinayaka Missions Medical College and Hospital (VMRF-DU), Keezhakasakudy, Puducherry, India. 2. Associate Professor, Department of Pharmacology, Mahatma Gandhi Medical College and Research Institute, Puducherry, India. 3. Assistant Professor, Department of Pharmacology, Vinayaka Missions Medical College and Hospital (VMRF-DU), Keezhakasakudy, Puducherry, India. 4. Associate Professor, Department of Pharmacology, Panimalar Medical College and Research Institute, Varadharajapuram, Poonamallee, Chennai, Tamil Nadu, India. 5. Professor, Department of Pharmacology, Sri Manakula Vinayagar Medical College and Hospital, Madagadipet, Puducherry, India. 6. Professor, Department of Pharmacology, Bhaarath Medical College, Chennai, Tamil Nadu, India.
 
Correspondence Address :
Balaji More,
Mahatma Gandhi Medical College and Research Institute, Puducherry, Puducherry,
Puducherry, India.
E-mail: drbdmore@gmail.com
 
ABSTRACT
: Metabolic Syndrome (MetS) is a complex of metabolic disorders which include obesity, insulin resistance, hyperglycaemia, hypertension along with dyslipidemia. It is not clear whether rosuvastatin is having role in new onset diabetes.

Aim: To assess the effect of rosuvastatin in comparison to glibenclamide on blood sugar levels and insulin resistance in diabetic rats.

Materials and Methods: This experimental study was carried out at Sri Manakula Vinayagar Medical College and Hospital, Kalitheerthalkuppam, Puducherry on 42 adult male Wistar rats, for 8 weeks. Diabetes was created by feeding rats with High Fat and High Sugar (HFHS) diet over a period of 8 weeks. In total, 42 rats were allocated to in 7 groups, consisting of 6 animals per group. Group I -vehicle treated animals (control), group II - HFHS diet animals (Diabetic Control- DC), group III- glibenclimide (G) (5 mg/kg) + HFHS, group IV-rosuvastatin (R) (5 mg/kg) + HFHS, group V-R (10 mg/kg) + HFHS, group VI-G (5 mg/kg) + R (5 mg/kg) + HFHS diet, group VII-G (5 mg/kg) + R (10 mg/kg) + HFHS diet continued for 4 weeks. Rosuvastatin and glibenclamide were dissolved in distilled water and 95% ethanol respectively, and were given orally daily for 4 weeks after induction of diabetes. Data were subjected to one-way ANOVA using Statistical Package for the Social Sciences (SPSS) version 24.0 followed by non parametric test with significance level set for p-value<0.05.

Results: By end of 8th week, on comparison of control group with HFHS diet and glibenclamide treated group, there was significant rise in body weight, serum Low Density Lipoprotein (LDL), in addition to Very Low Density Lipoprotein (VLDL) and Triglycerides (TG) levels. Glibenclamide treated (group III) animals on comparison with groups IV, V, VI, VII revealed reduction in body weight, serum LDL, VLDL and TG levels (p <0.001). Glibenclamide treated (group III) on comparing with groups IV, V, VI, VII, there was significant high serum HDL level (p<0.001). By 8th week, significant decrease in serum insulin and HOMA-IR level was observed when glibenclamide group III was compared with groups IV to VII treated animals (p <0.001).

Conclusion: The study results indicate that the combination therapy of rosuvastatin and glibenclamide demonstrate beneficial effects on weight reduction, lipid profile and insulin resistant.
Keywords : Cholesterol, Diabetes, Insulin resistance, Lipid profile
 
INTRODUCTION
Metabolic syndrome (MetS) consists of metabolic disorders which include obesity, insulin resistance, hyperglycaemia, hypertension along with dyslipidemia which is on a rise worldwide (1). Its prevalence is high especially among Asians including Indians. In India, it ranges from 16.3% to as high as 48.2%. It is critical risk factor accountable for Type 2 Diabetes Mellitus (T2DM) and Cardiovascular Diseases (CVD) which are responsible for increased mortality and morbidity (2).

Most subjects with MetS receive statins to manage dyslipidemia. Statins act by inhibiting of 3-hydroxy-3-methylglutaryl co-enzyme A (HMG-CoA) reductase. They are used to treat the dyslipidemia component of MetS. Additionally they demonstrate several pleiotropic actions like improvement in endothelial function in atherosclerosis (3),(4). Several clinical studies ascertained that statin are beneficial in reduction of atherosclerosis and have favourable influence in diabetic patient (4). On the contrary, statins are also considered to have role in new on-set T2DM, although uncertain by reducing the insulin secretion (5).

A recent meta-analysis revealed amplified risk of development of diabetes with statins intake (5). The proposed mechanism for the augmented risk of diabetes with statins because of enhanced formation of plasma-derived LDL cholesterol as a result of statin-induced antagonism of cholesterol production which results in direct inflammatory changes and oxidation in beta cell, in turn resulting in cellular apoptosis and impaired insulin secretion. Statins also exert influence on glucose metabolism and Insulin Resistance (IR) the probable mechanism could be reduction in insulin secretion (6),(7). The other likely causes of pathogeneses includes the effect of statins in HMG-CoA reductase inhibition, calcium release, isoprenoid synthesis, glucose transport, calcium mediated pancreatic insulin secretion, lowering of different isoprenoids (8). Hence, it is uncertain whether statins can certainly control Blood Sugar Levels (BSL) in diabetic patients, or even in the diabetic model animals.

Since statins are commonly used in prevention of CVD in diabetes and non diabetic patients, it is important to evaluate the effect of statin intake BSL and insulin resistance. Current therapeutic strategies for T2DM are limited and it involves insulin and oral hypoglycaemic drugs. Sulfonyl ureas are rapidly acting class of drugs which stimulate pancreatic insulin secretion e.g., Rosuvastatin, Glibenclimide, Glipizide. Rosuvastatin is commonly used stating to control dyslipidemia and reduce the risk of CVD. Therefore, this experimental study was carried out to evaluate effectiveness of rosuvastatin in controlling the diabetes and IR in diabetic rats as compared to glibenclimide.
 
MATERIAL AND METHODS
This observer-blinded experimental study was carried out at Sri Manakula Vinayagar Medical College and Hospital, Kalitheerthalkuppam, Puducherry, between 2016 to 2019. The study was carried out after the protocol was approved by Institutional Animal Ethics Committee (Approval letter No. VMMCH/2016/IAEC/21). Adult male Wistar rats of age 10-12 weeks and weight 120-180g were obtained from the King’s Institute Guindy, Chennai. Unhealthy animals were excluded from the study.

Procedure

The experiment was carried out in the light hours between 10.00 and 12.00 hours. The animals were cared and maintained as per Committee for the Purpose of Control and Supervision of Experiments on Animals guidelines and Good Laboratory Practice (GLP).

Animals were be given HFHS diet daily for a period for 8 weeks to induce diabetes. Rats were administered high fat by mixing [commercially available Vanaspati ghee (vegetable oil) and coconut oil] in the ratio of 3:1 (v/v). The dose given was 3mL/kg body weight/day oral diet and high sugar diet consisting of 25% fructose (which means that for each 100 mL water, 25 gram of fructose was added) in drinking water and fed over 24 hrs for 8 weeks. Rats were weighed prior, during and at the end of the experimental procedure. Total duration of study was six weeks (9).

Following chemicals and drugs were used as high fat (mixing vegetable oil + coconut oil of 3:1 ratio)- 3 mL/kg and high sugar; Diet (25% fructose for 24 hrs along with water), Glibenclimide (G) (5 mg/kg/day), Rosuvastatin (R) (5 and 10mg/kg/day), 95% ethanol, distilled water and appropriate vehicle will be used for control animals.

Six mice per group are sufficient for both the statistical significance and adherence to the rule of investing mice in experimentation. In total, 42 animals are required for test and they were divided into 7 groups consisting 6 animals as follows:

Group I: Vehicle treated animals (control)
Group II: HFHS diet animals - DC
Group III: G (5 mg/kg) +HFHS
Group IV: R (5 mg/kg) +HFHS
Group V: R (10 mg/kg) +HFHS
Group VI: G (5 mg/kg) +R (5mg/kg) +HFHS diet
Group VII: G (5 mg/kg) +R (10mg/kg) +HFHS diet

Rosuvastatin and glibenclamide were dissolved in distilled water and 95% ethanol respectively, and were given orally daily for 4 weeks after induction of diabetes and were continued for 4 weeks weeks.

Experimental procedure: The drugs and diet were given for a period of 8 weeks. On 4th and 8th week of experiment after overnight fasting, rats were given anaesthesia with small quantity of ether. Blood was obtained through retro-orbital puncture technique. Blood glucose, lipid profile and insulin levels were measured using the blood sample (10).

Blood glucose was measured with Accu-Chek (R) glucometer (by Roche Diabetes Care, Inc.,) after the collection of blood sample from the all night (12-15 hr) fasted rats on every week of experiment. Body weight gain or loss in each experimental rat were measured and recorded on every week with triple balance (9). Following lipid parameters were measured and recorded on 4th and 8th week

i. Serum Total Cholesterol (TC)
ii. Serum Triglycerides (TG)
iii. Serum High Density Lipoproteins (HDL)
iv. Serum Low Density Lipoproteins (LDL)
v. Serum Very Low Density Lipoproteins (VLDL)

Fasting serum insulin measurement and Homeostasis Model Assessment-estimated Insulin Resistance (HOMA-IR) were calculated.

Statistical Analysis

Data were subjected to one-way ANOVA which was followed with Post-Hoc Turkey’s test using SPSS software version 24.0 followed by non parametric test with significance level set for p-value <0.05.
 
RESULTS
The effect of rosuvastatin and glibenclamide (alone and in combination) on body weight in euglycemic and diabetes induced male albino Wistar rats are summarised in (Table/Fig 1).

On 0th week and at end of 1st week significant differences was not observed in weight of rats on comparison of control with different groups. By end of 8th week, when control (group I) was compared with HFHS diet (group II) and glibenclamide treated (group III) there was significant rise in body weight (p<0.001). When glibenclamide (5 mg/kg) treated animals (group III) compared with groups IV, V, VI, VII showed significant decrease in body weight (p <0.001).

The effect of rosuvastatin and glibenclamide (alone and in combination) on serum Fasting Blood Sugar (FBS) level in euglycemic and diabetes induced male albino Wistar rats have been summarised in (Table/Fig 2).

At 0th week and at the end of 1st week, significant changes were not observed in serum fasting BSLs. At end of 8th week when on comparison of normal control (group I) with group- II,III,IV,V,VI,VII significant rise in FBS level (p<0.001) was observed. There was statistical decrease in blood glucose level on comparing HFHS diet animals (group II) with groups III, IV, V (p<0.001, p<0.01, p<0.001). However, on comparison of glibenclamide (5 mg/kg) group of animals with groups IV, V, VI, and VII there was statistical reduction in serum FBS level was observed (p<0.05, p<0.05, p<0.01, p<0.001) respectively.

The effect of rosuvastatin and glibenclamide (alone and in combination) on lipid profile in euglycemic and diabetes induced male albino Wistar rats is summarised in (Table/Fig 3).

Serum Total Cholesterol level (TC): By end of 4th week, significant rise in serum TC level was observed on comparison of normal control (group I) with groups II to VII (p<0.001).

By end of 8th week, on comparison of normal control (group I) with groups II, III, IV, V, VI, VII treated rats there was significant rise in levels of serum TC (p<0.001). However, significant reduction in serum TC level was observed in groups III, IV, V, VI, VII of animals (p<0.001) when compared with groups II animals. On comparison of glibenclamide (5 mg/kg) treated (group III) with other groups IV, V, VI, VII treated animals there was significant reduction in TC level (p<0.001).

Serum Triglyceride level (TG): By end of 4th week, significant rise in serum TG level was observed in normal control (group I) compared to groups II, III, IV, V, VI, VII treated animals (p <0.001). Comparison of HFHS diet (group II) with group-III, IV, V, VII showed significant increased levels of serum TG (p<0.01, p<0.01, p<0.05, p<0.01) respectively, however, when glibenclamide (5 mg/kg) (group III) was compared with groups V, VI, VII showed significant increase in serum TG level (p<0.05, p<0.01, p<0.001) respectively.

On 8th week, when normal group compared with groups II, III, IV, V, VI, VII treated animals shows statistically significant increase in serum TG level (p<0.001). However, HFHS diet (group II) when compared with groups III, IV, V, VI, VII there was significant decrease levels of serum TG level (p <0.001). Glibenclamide (5 mg/kg) treated (group III) on comparing groups IV, V, VI, VII treated animals there was significant reduction in serum TG level.

Serum HDL level: On 4th week, observed a significant decrease in serum HDL level when compared normal (group I), with groups II to VII treated animals. Glibenclamide (5 mg/kg) on comparing with group V treated animals there was slight increase in the HDL level (p<0.05).

On 8th week, when normal group compared with group- II, III, IV treated animals shows statistically significant decrease (p<0.001) and groups V, VII showing an increase in serum HDL level (p<0.001). However, HFHS diet when compared with groups III to VII there was significant increased levels of serum HDL level (p<0.001). Glibenclamide (5 mg/kg) treated (group III) on comparing groups IV to VII treated animals there was significant high serum HDL level (p<0.001).

Serum VLDL: At the end of 4th week there was significant rise in VLDL level when compared control (group I) with groups II, III, IV, V, VI, and VII treated animals (p <0.001). On comparing HFHS diet group animals with groups III, V, and VII observed significant increase in serum VLDL level (p<0.05, p<0.001, p<0.05). Also observed significant increased levels of VLDL when compared glibenclamide (5 mg/kg) with groups VI, VII (p<0.001).

At the end of 8th week, VLDL level was significantly increased groups II, III, IV, V, VI, and VII treated animals when compared with normal group (p<0.001). But observed a significant reduced levels of VLDL on comparison of HFHS diet (group II) with groups III, IV, V, VI, VII treated animals (p<0.001). Also there was significant decrease in VLDL level with groups IV, V, VI, and VII when compared with glibenclamide (5mg/kg) treated group (p<0.001).

Serum LDL level: At the end of 4th week there was significant increase in LDL level when compared control with groups II, III, IV, V, VI, VII treated animals (p<0.001). HFHS diet (group III) animals on comparing with groups III, IV, VII there was significant raised levels of serum LDL (p<0.01, p<0.01, p<0.001). When compared glibenclamide group (5 mg/kg) with groups V, and VII treated animals there was significant increase in the LDL level (p<0.001).

At the end of 8th week, when serum LDL level was compared between control with groups II, III, IV, V, VI, VII there was significant rise in serum LDL value (p<0.001). But observed a significant reduction in LDL level on comparing group II with groups III, IV, V, VI, and VII (p<0.001). Also there was significant decrease in LDL level with groups IV, V, VI, and VII when compared with glibenclamide (5mg/kg) treated group (p<0.001).

On 4th week there was significant rise in serum insulin level and HOMA-IR on comparison of normal control with groups II to VII treated animals (p<0.001). HFHS diet group animals when compared with groups VII observed significant increase in serum insulin level (p<0.01). However, HOMA-IR level were increased with groups IV, V, VII (p<0.001, p<0.05, p<0.01). Glibenclamide (5 mg/kg) group when compared with group VI and group IV there was significant increase in the insulin [(p<0.01) and (p<0.05), respectively] and HOMA-IR (Table/Fig 4).

By end of 8th week, there was significant increased levels of insulin and HOMA-IR level when compared control with groups II to VII treated animals (p<0.001). When compared HFHS diet animals with groups III to VII treated animals there was significant reduction in insulin level and HOMA-IR (group I) Significant decrease in serum insulin and HOMA-IR level was observed when glibenclamide (5 mg/kg) was compared with groups IV to VII treated animals (p<0.001) (Table/Fig 4).
 
DISCUSSION
Metabolic syndrome has become a global epidemic. The threat diabetic complications can be reduced substantially if properly managed (10). The association of dyslipidemia and heart disease is well established. Moreover reduction of levels of harmful lipids satisfactorily decreases morbidity and mortality in CHDs. The concomitant treatment with glibenclamide and rosuvastatin is a rationale approach in the management of diabetes and reducing associated risk factors especially in management of CVD (4). Hence this research project was conducted to determine the effect of rosuvastatin on BSL and IR in HFHS diet induced diabetic male albino Wistar rats.

In this study, MetS like state was induced in rats by administration of HFHS prepared with mixture of vegetable oil with coconut oil. These ingredients are commonly used by a large population to cook food, which could be responsible for developing hyperlipidemia. High sugar diet was given to rats as 25% fructose. As compared to glucose, dietary fructose is stronger inducer of insulin resistance, impairment of glucose tolerance, high insulin levels, abnormal lipid levels, hypertriglyceridemia, and increased blood pressure (11). Animal model with hyperlipidemic and insulin resistant states that imitated the pathogenesis of clinical effects as observed in patients as a result of imbalanced diet intake was developed by feeding HFHS diet to rats (12). In this study, animal model was developed by feeding HFHS diet for 8 weeks to rats, which showed a significant rise in their injurious serum lipid and lowering of high density lipoproteins cholesterol. Additionally high Fasting Blood Glucose (FBG), insulin and HOMA-IR values was also seen observed representing a MetS.

Effect on Blood Sugar levels: The HFHS (DC) group has demonstrated a rise in the FBG. There was a significant rise in FBG at the end of 8th week in comparison with control group was the proof of induced hyperglycaemia as in MetS. All the treated groups demonstrated a decline in BSL indicating their blood glucose lowering action.

There was significant raise in blood glucose level in all treated groups up to four weeks when compared with control group. This study results are in congruence with results of Munshi RP et al., indicating that we could achieve the development of diabetic rat model by feeding HFHS (12).

At the end of 8th week there was significant rise in FBG in DC animals when compared with other (groups –III, IV, and V). The FBG level increased in both the doses of rosuvastatin treated groups as monotherapy when compared to glibenclamide (5mg/kg) treated animals (p<0.05). As expected there was statistical increase in FBG in glibenclamide group as compared to DC (p<0.001). Our results are in congruence with findings of study conducted by Sokolovska J et al., wherein they found glibenclamide lowered the BSL and raised the insulin in similar rat models (13). In a previous study by George AV et al., long-term dose of glibenclamide lowered BSL, liver glycogen and protein. It also raised hepatic and serum lipids and liver organic phosphates in normal rats. Accumulation of lipids result in significant rise in weight of glibenclamide treated rats (14).

This study demonstrated that glibenclamide caused significant fall in BSL in rats with HFHS induced diabetes. Rosuvastatin did not cause any significant change of this parameter. Moreover combination treatment more effective for controlling BSL as compared to glibenclamide alone in rats with long-term HFHS induced diabetic. Nevertheless, more studies are required to elucidate the actual mechanism of this synergistic effect with this combination of drugs. Our results are in congruence with results of the earlier study results (8),(11),(13). Statins did not alter the BSL and plasma insulin induced as a result of glucose administration. There was no improvement in glucose intolerance observed during Oral Glucose Tolerance Test (OGTT) in long-term treatments of GK rats with statins in a previous literature research (15).

Statins impacts glucose metabolism in several ways. They activate Endogenous Glucose Production (EGP) by up-regulation of gluconeogenic genes hepatic cells. 16 statins up regulate the cytoplamic X receptor (PXR) which stimulates expression of proteins responsible for hepatic glucose and lipid metabolism (16),(17). However, in-vivo experimentation to assess the effectiveness of statin on glucose metabolism in type 2 diabetes patients demonstrated insignificant action on EGP. There was no change in basal EGP in type 2 diabetics in atorvastatin (10 mg for 12 weeks) or simvastatin (80 mg/day for 8 weeks) therapy (18),(19).

In an experimental study conducted in Wister rats, high BSL were produced injecting intraperitoneal low dose (25 mg/kg) streptozotocin. Rosuvastatin produced less impairment of glucose tolerance as compared to atorvastatin. Therefore, in patients with high risk with diabetes or having impaired glucose tolerance rosuvastatin could be a better choice for preventing and treating dyslipidemia (20). There was no effect seen with rosuvastatin on rise in BSL and loss of body weight. However, one of the previous studies showed that treatment with pravastatin and olmesartan resulted in synergistic improvement in glucose intolerance by improving increased glucose uptake by the tissue. The effect seems to be produced by improvement in insulin sensitivity by reducing oxidative stress (21).

In patients with diabetes, statin produced a modest increase in HbA1c (22). As a result for diabetic patients on statin should receive greater intensification of treatment. Nevertheless, statins with lower potency have been less pronounced effect on HBA1c (23).

Genetic research is required to identify the role gene variants in the target genes for statins which could be responsible for increasing the risk of T2DM. As compared to clinical trials, population-based studies indicate relatively increased incidence of T2DM in subjects treated with statin. Emerging newer data indicates that pravastatin is the least diabetogenic among statins. Even though statins has diabetogenic potential, the consensus is in favour of use of statins as lowering cardiovascular events definitely outweigh the risk of diabetes (24),(25). As the potential for diabetogenicity varies among statins, it is advisable to personalise statin treatment by categorising patients who are less risk of T2DM due to statins with lower diabetogenic potential (26).

Effect on insulin levels: By the end of 4 weeks there was significant rise in serum insulin and HOMA-IR level in all groups compared to control group, whereas at the end of weeks significant decrease in serum insulin and HOMA-IR level was observed with group- [IV, V, VI, VII] treated rats (p<0.001), in comparison to glibenclamide (5 mg/kg). Animals which were administered glibenclamide showed group significantly reduced HOMA-IR values (p<0.001). All groups treated with sulphonyl urea drugs showed increase insulin secretion in diabetic rats which was in congruence with earlier studies (27).

In one of the experimental studies, rosuvastatin treatment groups have increased insulin sensitivity in whole body and peripheral tissue by enhancing cellular insulin signal transduction (28). In another study with rats models, demonstrated that pravastatin and atorvastatin had paradoxical effects on insulin sensitivity and vitamin D3 levels. Pravastatin increased insulin sensitivity by elevation of 1,25-(OH)(2)-D3 whereas atorvastatin produced decrease in insulin sensitivity which was independent of 25-OH-D3 levels (29). In another study with mice model atorvastatin produced improvement in glucose metabolism by enhancing insulin sensitivity. Northern blot testing showed reduction in levels of mRNA of sterol regulatory element binding protein-1 (SREBP-1) and glucose-6-phosphatase (G6Pase). This may be responsible for improving glucose metabolism and insulin sensitivity (30).

Another study reveals that rosuvastatin improves insulin sensitivity rats fed with HFD. The mechanisms involved are reduction of leptin and enhancement of Sirtuin 1, PPAR-γ and GLUT-4 expression in white adipose tissue. Sirtuin 1 is a mediator of rosuvastatin on insulin sensitivity in overweight rats induced by diet (31).

Effect on body weight: The treatment with combination of rosuvastatin and glibenclamide has significantly reduced body weight in comparison with HFHS control (DC) group than either of them as monotherapy. Similar reduction in body weight with glibenclamide was observed in clinical study by Martin S et al., (22).

Effect on lipid profile: By the end of 4th week significant decrease in serum total TC, TG, LDL and VLDL level was observed in control group when compared other groups of animals. At the end of 8th weeks, on comparing glibenclamide (5 mg/kg) treated group with other groups [IV, V, VI, VII], there was significant reduction in TC, LDL, VLDL,TG and increase in HDL levels which correlates with previous study results by Asad M et al., (23). Rosuvastatin has significant pleiotropic actions which are secondary to inhibition of oxidative stress and reduction in advanced glycation end products (AGEs) accumulation. These actions may provide potential benefits in apart from the lipid lowering in the management of diabetes (24).

Under different physiological conditions the factors that affect metabolism of glucose may also have action metabolism on lipid. It is found that cholesterol and triglyceride levels are also increased significantly in T2DM (32),(33). Our findings suggest that rosuvastatin alone as monotherapy improved lipid profile while glibenclamide did not have effect on lipid profile even by 8 weeks which is in congruence with results of earlier study (34).Previous studies have shown high levels of triglycerides observed in long duration diabetic rats was because to disturbance in the triglyceride removal mechanism. Simvastatin decreased plasma triglyceride levels in rats both by enhancing triglyceride removal and by decreasing its entry into the blood (34).

Ali H A et al., has described majoralteration in the lipid parameters with reduction in TC, LDL VLDL cholesterol and TG levels and increase in HDL cholesterol on treatment with both atorvastatin and pioglitazone (11). In our study also the glibenclamide, rosuvastatin and their combination also demonstrated their property of correcting hyperlipidemic and improving insulin sensitivity. Maximum benefit was seen with high dose rosuvastatin (10 mg/kg) as monotherapy alone followed by combination along with glibenclamide.

An experimental study in Wistar rats with streptozotocin induced hyperglycemia revealed that hydrophilic rosuvastatin caused minor impairment in glucose tolerance. Therefore, rosuvastatin would be a preferred choice than lipophilic atorvastatin for management dyslipidemia especially in subjects with impaired glucose tolerance or at risk of developing diabetes (20).

Limitations

Chronic intake of HFHS diet produced peroxidation of lipids and abnormalities in antioxidant mechanisms in the tissues. It also causes slowing in gastric transit time, so food remains in stomach for longer duration than required for otherwise healthy digestion. Antioxidant activities are produced by enzymatic and nonenzymatic mechanisms which were not evaluated in this study. The effects of combination therapy (glibenclamide and rosuvastatin) on the liver function parameters were not studied. High blood glucose produces increased oxidative reactions and release of free radical along with increase in alanine transaminase and aspartate transaminase levels. By studying the effects of combination therapy (glibenclamide and rosuvastatin) on the liver function parameters more information could have been found on the hepatoprotective actions. Only two doses were studied. Duration of the study was short. Hypertension parameter was not explored which is major criteria in MetS. No diabetic limitations were set in this study before giving treatment, but the groups treated with HFHS diet showed increase in blood glucose of diabetic level as in previous study limits.
 
CONCLUSION
High fat high sugar diet induced diabetes in rats is well-accepted animal model. The combination therapy of rosuvastatin and glibenclamide demonstrated significant weight reduction, antihyperglycemic and anti lipidemic activity which represents to be an effective drug combination for adequate glycemic control and prevention of complications in the management of diabetes.
 
ACKNOWLEDGEMENT
We would all the technician and animal house staff for taking care of the animals.
 
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