Management of Diabetic with Combined Therapy of Reducdyn and Metformin in Streptozotocin induced Diabetic Male Rats.

 

Nwauche Kelechi T., Monago Comfort C.* and Onwuka Franscis.

Department of Biochemistry, Faculty of Chemical Sciences, College of Natural and Applied Sciences, University of Port Harcourt, Choba, Rivers State, Nigeria.

 

 

ABSTRACT:

Metformin is a known antidiabetic drug, while reducdyn is a known liver drug. Diabetes mellitus is always associated with liver problem. The present study was designed to investigate the effects of metformin and reducdyn in management of diabetes mellitus and its liver associated problems in male rats. Metformin was administered at a dose of 1.4mg/kg body weight per day for the period of the treatment. This fixed dose of metformin was co-administered differently with reducdyn at different doses of 0.25mg/kg and 0.5mg/kg body weight to groups IV and V rats respectively for 3, 6 and 9 weeks. The statistical analysis was carried out using one way ANOVA followed by post hoc LSD multiple comparison on SPSS 19. Metformin, when administered alone and its co-administration with the different doses of reducdyn showed significant decrease in blood glucose concentration for 3, 6 and 9 weeks of treatment when compared with the diabetic control group. There was no significant (P>0.05) difference in the plasma AST and ALT activities for the first 3, 6 and 9 weeks of treatment in all the groups when compared with the diabetic control group. There was a significant (P˂0.05) decrease in AST activities of groups III and IV (DCR on metformin (1.4mg/kg) and DCR on metformin (1.4mg/kg) + reducdyn (0.25mg/kg) body weight) after 6 weeks of treatment when compared with the Normal control (Group I). After the 9th week of treatment, it was observed that only group III had a significant (P˂0.05) decrease in the plasma AST and ALT activity when compared with the normal control group. It is also noticed that groups III and IV had an appreciable reduction rate in the plasma AST activities when compared with the diabetic and normal control groups. There was a significant (P˂0.05) decrease in the plasma ALP activities in all the groups treated for the first 3 weeks when compared with the normal and diabetic control groups (groups I and II) except for groups III. However, there was no significant (P>0.05) difference in the enzyme activities between the two control groups after 3 weeks of treatment. After 6 weeks groups IV and V had a significant decrease in the plasma ALP activities when compared with the normal and diabetic control groups (groups I and II). Group III had no significant (P˂0.05) difference in the plasma ALP activities when compared with the two control groups. As the treatment progressed to the 9th week, there was no significant (P˂0.05) difference in the plasma ALP activities between the normal and diabetic control groups. Only group V had a significant (P˂0.05) decrease in the plasma ALP activities when compared with the normal control group. Groups III and IV, had no significant (P˂0.05) difference in the plasma ALP activities when compared with the normal control group and diabetic control group (groups I and II).

 


In histological evaluation of the pancreas andhepatic tissue, the islet cells of the Langerhan of the pancreas were repopulated and preserved in all the groups treated with the drugs against the diabetic control group whose islet cells were reduced, while mild inflammation of the hepatic tissue was observed in all the groups throughout the period of treatment, except for those treated with metformin alone whose hepatic tissues were seen to have periportal inflammation after 3 and 6 weeks of treatment.

 

KEY WORDS: Diabetes Mellitus, Liver enzymes. Strepzotocin, Metformin, Reducdyn

 

INTRODUCTION:

The liver helps maintain normal blood glucose concentration in the fasting and postprandial states. Loss of insulin effect on the liver leads to glycogenolysis and an increase in hepatic glucose production. Abnormalities of triglyceride storage and lipolysis in insulin-sensitive tissues such as the liver are an early manifestation of conditions characterized by insulin resistance and are detectable earlier than fasting hyperglycemia. The precise genetic, environmental, metabolic factors and sequence of events that lead to the underlying insulin resistance, however, is not fully understood (Lewis et al., 2002)

 

In animal models, chronic hyperinsulinemia is found to predispose the liver to relative resistance to insulin. This is characterized by a failure of insulin to signal an increase in insulin receptor substrate-2. Upregulation of sterol regulatory element-binding protein 1c (SREBP-1c) also occurs, leading to increased lipogenesis (Shimomura et al., 2000). Despite down-regulation of the insulin receptor substrate-2-mediated insulin signaling pathway in insulin-resistant states, the up-regulation of SREBP-1c and subsequent simulation of de novo lipogenesis in the liver leads to increased intracellular availability of triglycerides, promoting fatty liver. This also increases VLDL assembly and secretion (Lewis et al., 2002). Thus, hyperinsulinemia might directly lead to hepatic insulin resistance with associated fatty changes.

 

The excess in free fatty acids found in the insulin-resistant state is known to be directly toxic to hepatocytes. Putative mechanisms include cell membrane disruption at high concentration, mitochondrial dysfunction, toxin formation, and activation and inhibition of key steps in the regulation of metabolism (Neuschwander-Tetri and Caldwell 2003).  Other potential explanations for elevated transaminases in insulin-resistant states include oxidant stress from reactive lipid peroxidation, peroxisomal beta-oxidation, and recruited inflammatory cells. The insulin-resistant state is also characterized by an increase in proinflammatory cytokines such as tumor necrosis factor-α (TNF-α), which may also contribute to hepatocellular injury. In preliminary studies, an increased frequency of specific TNF-α-promoter polymorphism was found in nonalcoholic steatohepatitis (NASH) patients, suggesting a possible genetic link or predisposition to fatty liver found in insulin-resistant states (Grove, 1997)

 

The above theories all attribute elevated transaminitis to direct hepatocyte injury. It is also hypothesized that elevation in ALT, a gluconeogenic enzyme whose gene transcription is suppressed by insulin, could indicate impairment in insulin signaling rather than purely hepatocyte injury (O'Brien and Granner 1991).

 

Individuals with type 2 diabetes have a higher incidence of liver function test abnormalities than individuals who do not have diabetes. Mild chronic elevations of transaminases often reflect underlying insulin resistance. Elevation of transaminases within three times the upper limits of normal is not a contraindication for starting oral antidiabetic or lipid-modifying therapy. In contrast, antidiabetic agents have generally been shown to decrease alanine aminotransferase levels as tighter blood glucose levels are achieved.

 

Liver function tests (LFTs) are commonly used in clinical practice to screen for liver disease, monitor the progression of known disease, and monitor the effects of potentially hepatotoxic drugs.

 

The most common LFTs include the serum aminotransferases and alkaline phosphatase. Aminotransferases, such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST), measure the concentration of intracellular hepatic enzymes that have leaked into the circulation and serve as a marker of hepatocyte injury. Alkaline phosphatase (ALP), γ-glutamyl transpeptidase (GGT), act as markers of biliary function and cholestasis. Chronic mild elevation of transaminases are frequently found in type 2 diabetic patients.

 

This study is designed to evaluate the effect of the combined therapy of metformin and reducdyn on diabetic induced liver problems in male rats.

 

MATERIALS AND METHOD:

Drugs and Equipment.

Glucophage (metformin), and reducdyn were obtained from Dooka Pharmarcy, Ltd opposite the main gate of University of Port Harcourt teaching hospital, Alakahia, Port Harcourt while Streptozotocin was obtained from glaxosmithline ltd Ibadan. All other reagents were of analytical grade. Optical densities were measured using digital spectrophotometer (model 752S).

 

Animals

Adult male wistar albino rats weighing 170-200g were obtained from the animal House of the Department of Biochemistry, University of Port Harcourt, Port Harcourt, Nigeria. The rats were randomly sorted into five groups of nine animals each (these nine animals were subdivided into three groups of three animals each for the 3, 6 and 9 weeks of treatment) so that the average difference was ± 2.0g. The animals were housed in plastic cages. The animals were allowed normal feed i.e guinea growers mash (Port Harcourt Flour Mills, Port Harcourt, Nigeria) and water ad libitum.

 

Experimental Design

The rats were divided into five groups of nine rats each after, the induction of streptozotocin diabetes.

Group-I--Normal control rats

Group-II --Diabetic control rats

Group-III-- Diabetic rats received a standard drug, GlucophageTM Metformin (1.4mg/kg)

Group-IV-- Diabetic rats received 1.4mg/kg of GlucophageTM (metformin) and 0.25mg/kg of Reducdyn.

Group-V--Diabetic rats received 1.4mg/kg of GlucophageTM (metformin) and 0.5mg/kg of Reducdyn.

 

Anti-Diabetic Study/Drug Administration

The rats were randomly sorted into five groups of nine animals each (these nine animals were subdivided into three groups of three animals each for the 3, 6 and 9 weeks of treatment) so that the average difference was ± 2.0g. The animals were housed in plastic cages. After a one week acclimatization period on guinea growers mash (Port Harcourt Flour Mills, Port Harcourt, Nigeria), the animals were fasted overnight and diabetes was induced by intraperitoneal injection of freshly prepared solution of streptozotocin (160mg/kg body weight) in distilled water, while the normal control rats (NCR) were injected with distilled water alone. Seven days after administration of streptozotocin, the animals were again fasted and blood collected via tail cutting (Burcelin et al., 1995), for the determination of their fasting glucose levels. Then the rats were kept for 3 days to stabilize the diabetic condition (Jyoty et al., 2002) before commencing treatment, which lasted for 9 weeks.

 

The first and second groups (normal control rats (NCR) and diabetic control rats (DCR) received appropriate volume of water using a gavage via intubation. The third group i.e the first treatment group received daily by intra-gastric gavages, 1.4mg/kg body weight of GlucophageTM (metformin); the fourth group received 1.4mg/kg body weight of  GlucophageTM  (metformin) and 0.25mg/kg body weight of Reducdyn TM  via the same route and the fifth group received 1.4mg/kg body weight of  GlucophageTM  (metformin) and 0.5mg/kg body weight of ReducdynTM  via the same route.

 

The animals were allowed normal feed and water ad libitum. At the end of each treatment period, i.e., 3 week, 6 week, and 9 week, the rats were weighed, fasted overnight and their fasting glucose level estimated by an automatic one touch glucometer. They were anaesthetized by exposure to chloroform. While under anesthesia, they were painlessly sacrificed and blood was collected from each rat into heparin sample bottles. The heparin anti-coagulated blood samples were centrifuged at 1000 x g for 10 min, after which their plasma was collected and stored for subsequent analysis.

 

Liver Function Tests

Aspartate aminotransferase (AST), Alanine aminotransferase (ALT) and Alkaline phosphatase (ALP) activities were determined by enzymatic methods with commercial test kits (Randox Laboratories, Crumlin, England).

 

Histopathology of the pancreas and liver.

On the last day of each of the study period, i.e 3, 6 and 9 weeks, the animals were sacrificed and quickly dissected.  Slices of pancreas and liver of the rats were fixed in 10 % formalin and used for histopathological studies.

 

Statistical Analysis of Data

Dataobtained were analyzed statistically by means of one-way ANOVA and post hoc LSD, on SPSS 19. In all, p<0.05 was considered significant. Data were presented as means.d (standard deviation).

 

RESULTS:

Table 1. below shows the results of the effect of co-administration of metformin and reducdyn onplasma glucose level of normal rats and streptozotocin induced diabetic male rats.

 


 

 

Table 1: Effect of co-administration of metformin and reducdyn on plasma glucose level of normal and streptozotocin induced diabetic male rats.

Group

Treatment

Serum glucose  level (mmol/l)

3 weeks

6 weeks

9 weeks

I

Normal Control Rats (NCR)

4.2±0.26a,b

3.9±0.85a,b

4.8±0.91a,b

II

Diabetic Control Rats (DCR)

8.1±0.32a,b

7.0±0.40a,b

6.3±0.43a,b

III

DCR on Metformin (1.4mg/kg)

4.6±0.97b

4.2±0.80b

4.5±0.20b

IV

DCR on Metformin (1.4mg/kg)  + Reducdyn (0.25mg/kg)

5.6±0.41a,b

3.8±0.20b

4.2±0.05b

V

DCR on Metformin (1.4mg)  + Reducdyn (0.5mg/kg)

6.3±0.77a,b

4.9±0.98b

4.4±0.98b

Values are expressed as Mean ±SD; n=3, per group/week. Values in the same column with common superscript letters (a,b,…) are significantly different at P>0.05.

Superscript A(a)  represents significant difference when group I (normal control rats) are compared with other groups at P˂0.05.

Superscript B(b)  represents significant difference when group II (diabetic control rats) are compared with other groups at P˂0.05.

Values without superscripts indicates no significant difference when compared with the normal and diabetic control groups (groups I and II)

 

Table 2: Effect of co-administration of metformin and reducdyn on plasma Aspartate transaminase (AST) activities of normal and streptozotocin induced diabetic male rats.

Group

Treatment

Enzyme Activity (Iu/L) (mean ±S.D)

3 weeks

6 weeks

9 weeks

I

Normal Control Rats (NCR)

21.6±2.30

25.6±6.11a

27.0±4.00a

II

Diabetic Control Rats (DCR)

19.0±0.00

21.6±2.30b

23.0±4.00b

III

DCR on Metformin (1.4mg/kg)

20.3±2.30

19.3±2.51a

18.3±5.03a

IV

DCR on Metformin (1.4mg/kg)  + Reducdyn (0.25mg/kg)

17.0±3.46

 18.3±5.03a

19.3±3.51

V

DCR on Metformin (1.4mg/kg)  + Reducdyn (0.5mg/kg)

20.0±9.64

23.0±4.00

27.0±4.00

Values are expressed as Mean ±SD; n=3, per group/week. Values in the same column with common superscript letters (a,b,…) are significantly different at P>0.05.

Superscript A(a)  represents significant difference when group I (normal control rats) are compared with other groups at P˂0.05.

Superscript B(b)  represents significant difference when group II (diabetic control rats) are compared with other groups at P˂0.05.

Values without superscripts indicates no significant difference when compared with the normal and diabetic control groups (groups I and II)

 

Table 3: Effect of co-administration of metformin and reducdyn on plasma Alanine transaminase (ALT) activities of normal and streptozotocin induced diabetic male rats.

Group

Treatment

Enzyme Activity (Iu/L) (mean ±S.D)

3 weeks

6 weeks

9 weeks

I

Normal Control Rats (NCR)

10.0±2.00

11.0±1.73

12.3±0.57a

II

Diabetic Control Rats (DCR)

8.0±2.00

10.0±3.00

11.6±1.52b

III

DCR on Metformin (1.4mg/kg)

7.3±2.30

8.0±1.00

8.6±1.15a

IV

DCR on Metformin (1.4mg/kg)  + Reducdyn (0.25mg/kg)

8.6±1.15

9.0±2.00

9.6±2.51

V

DCR on Metformin (1.4mg/kg)  + Reducdyn (0.5mg/kg)

7.6±4.04

8.6±1.15

9.6±2.08

Values are expressed as Mean ±SD; n=3, per group/week. Values in the same column with common superscript letters (a,b,…) are significantly different at P>0.05.

Superscript A(a)  represents significant difference when group I (normal control rats) are compared with other groups at P˂0.05.

Superscript B(b)  represents significant difference when group II (diabetic control rats) are compared with other groups at P˂0.05.

Values without superscripts indicates no significant difference when compared with the normal and diabetic control groups (groups I and II)

 

Table 4: Effect of co-administration of metformin and reducdyn on plasma Alkaline phosphatase (ALP) activities of normal and streptozotocin induced diabetic male rats.

Group

Treatment

Enzyme Activity (Iu/L) (mean ±S.D)

3 weeks

6 weeks

9 weeks

I

Normal Control Rats (NCR)

40.0±6.00a

66.0±10.00a

79.3±1.15a

II

Diabetic Control Rats (DCR)

40.0±2.00b

64.0±7.21b

76.6±4.16b

III

DCR on Metformin (1.4mg/kg)

32.6±2.08

56.6±5.03

69.3±4.61

IV

DCR on Metformin (1.4mg/kg)  + Reducdyn (0.25mg/kg)

28.3±4.61a,b

42.6±3.05a,b

64.0±7.21

V

DCR on Metformin (1.4mg/kg)  + Reducdyn (0.5mg/kg)

30.3±6.42a,b

45.3±1.15a,b

61.3±13.61a

Values are expressed as Mean ±SD; n=3, per group/week. Values in the same column with common superscript letters (a,b,…) are significantly different at P>0.05.

Superscript A(a)  represents significant difference when group I (normal control rats) are compared with other groups at P˂0.05.

Superscript B(b)  represents significant difference when group II (diabetic control rats) are compared with other groups at P˂0.05.

Values without superscripts indicates no significant difference when compared with the normal and diabetic control groups (groups I and II)

 

Fig. 1 Histopathological studies of the pancreas of normal and diabetic control rats after 3 weeks of treatment.

 

Fig. 2 Histopathological studies of the pancreas after 3 weeks of treatment with metformin alone and its combination with reducdyn.

 

 

Normal Control Rats after 6 weeks of treatment

 

Diabetic Control Rats After 6 Weeks of treatment  

SLID Slide Shows Reduced Islet

 

DCR treated with Metformin 1.4mg for 6 weeks Slide shows repopulation of islet cell

DCR treated with Metformin 1.4mg/kg and Reducdyn 0.25mg/kg for 6 weeks Slide shows increased islet cell mass

 

DCR treated with Metformin 1.4mg/kg and Reducdyn 0.5mg/kg for  6 weeksSlide shows repopulation of islet cells

 

Fig. 3: Histopathological studies of the pancreas after 6 weeks of treatment with metformin alone and its combination with reducdyn.

 

 

After 9 weeks of treatment

 

Normal control Rats after 9 weeks of treatment No Histologic change seen

Diabetic Control Rats After 9 Weeks of treatment SLID Slide Shows Reduced Islet cell mass

 

DCR treated with Metformin 1.4mg for 9 weeks Slide shows no change in islet cell mass

DCR treated with Metformin 1.4mg/kg and Reducdyn 0.25mg/kg  for  9 weeks Slide shows no histologic change in the islet cell mass

 

DCR treated with Metformin 1.4mg/kg and Reducdyn 0.5mg/kg for 9 weeks Slide shows normal islet cells

 

Fig. 4 Histopathological studies of the pancreas after 9 weeks of treatment with metformin alone and its combination with reducdyn.

 

 

Normal Control Rats after 3 weeks of treatment

 

DCR after 3 weeks of treatment Slide shows mild inflammatory changes of the hepatocytes

DCR treated with Metformin 1.4mg for 3 weeks Slide shows inflammatory change.

 

DCR treated with Metformin 1.4mg and reducdyn 0.25mg for 3 weeks Slide shows mild inflammatory change.

DCR treated with Metformin 1.4mg and reducdyn 0.5mg for 3 weeks Slide shows mild inflammatory change.

Fig. 5  Histopathological studies of the liver of normal and diabetic control rats after 3 weeks of treatment with metformin alone and its combination with reducdyn.

.

After 6 weeks of treatment.

 

 

 

DCR after 6 weeks of treatment Slide shows mild inflammatory changes of the hepatocytes

DCR treated with Metformin 1.4mg for 6 weeks Slide shows inflammatory change.

 

 

DCR treated with Metformin 1.4mg and reducdyn 0.25mg for 6 weeks Slide shows mild inflammatory change

DCR treated with Metformin 1.4mg and reducdyn 0.5mg for 6 weeks Slide shows mild inflammatory change.

 

Fig. 6: Histopathological studies of the liver of normal and diabetic control rats after 6  weeks of treatment with metformin alone and its combination with reducdyn

 

After 9 weeks of treatment                         

 

Normal Control Rats after 9 weeks of treatment Slide shows periportal inflammation

Diabetic Control Rats after 9 weeks of treatment Slide shows inflammatory changes

 

 

DCR treated with Metformin 1.4mg for 9 weeks Slide shows inflammatory change.

DCR treated with Metformin 1.4mg and reducdyn 0.25mg for 9 weeksSlide shows mild inflammatory change.

 

 

DCR treated with Metformin 1.4mg and reducdyn 0.5mg for 9 weeks Slide shows mild inflammatory change.

 

 

Fig. 7: Histopathological studies of the liver of normal and diabetic control rats after  9 weeks of treatment with metformin alone and its combination with reducdyn


 

DISCUSSION:

The success recorded in the use of streptozotocin (STZ) for the induction of diabetes mellitus through the administration of 160mg/kg body weight can be attributed to the work of Ferreira et al., 2002. This achievement was confirmed by evaluation of fasting blood glucose concentration.

 

Normal control rats maintained a fairly stable level of glucose throughout the study period with the values 4.2±0.26, 3.9±0.85 and 4.8±0.91 for 3, 6 and 9 weeks of treatment respectively. There was however significant (0.05) increase in the level of glucose concentration for the diabetic control rats reaching a hyperglycemic level of 8.1±0.32, 7.0±0.40 and 6.3±0.43 for 3, 6 and 9 weeks of treatment respectively (see table 1).

 

Group III treated with standard drug (metformin 1.4mg/kg) showed a significant (0.05) decrease in serum glucose levels on the 3rd, 6th and 9th week of treatment when compared with the diabetic control group (group II) as shown in table 1.

 

When metformin 1.4mg/kg was co-administered with different doses of reducdyn, there was a significant (0.05) decrease in the level of glucose at the 6th and 9th week of treatment than the first 3 weeks of treatment.

 

Aspartate amino transferase (AST), an enzyme involved in the transfer of amino group from aspartate to alpha keto acid is an established marker of hepatic function (Varley et al., 1980). Elevated level of this enzyme is usually interpreted as indicative of hepatic damage/destruction (Varley et al., 1980). From table 2 above, it will be observed that there was no significant (0.05) difference in the plasma AST activities in the first 3 weeks of treatment in all the groups when compared with the normal and diabetic control groups (group I and II). As the treatment continued to the 6th week, there was a significant (0.05) decrease of the AST activities of groups III and IV (DCR on metformin (1.4mg/kg) and  DCR on metformin (1.4mg/kg) + reducdyn (0.25mg/kg) body weight) when compared with the normal control group (group I). After the 9th week of treatment, it was observed that only group III had a significant (0.05) decrease in the plasma AST activity when compared with the normal control group. It is also noticed that groups III and IV had an appreciable reduction rate in the plasma AST activities when compared with the diabetic and normal control groups.

 

Alanine amino transferase (ALT), another enzyme used as marker of hepatic function had no significant (0.05) difference in its activity among the groups treated for 3 and 6 weeks respectively. As the treatment continued to the 9th week, it was observed that group III (DCR on metformin, 1.4mg/kg) decreased significantly (0.05) in plasma ALT activity when compared with the normal control group. From the above discussion, it will be observed that Glucophage (metformin), 1.4mg/kg body weight was observed to significantly lower the plasma enzyme activities of AST and ALT, an effect that appear quite confusing apparently. This is due to the fact that some oral hypoglycemic drugs, in addition to their established therapeutic effects, also possibly elicit some deleterious effects on the hepatocytes. This is in line with the suggestion of Katzung (1982) “that drugs in addition to their desired clinical effects also elicit some other effects, deleterious or otherwise on some metabolic/physiological system of the host.

 

However, it is possible that the drug, Glucophage may have possed these effects on the enzyme activity by having direct competitive effect on the enzyme.

 

Alkaline phosphatase (ALP), a hydrolase enzyme responsible for removing phosphate group from many types of molecules including nucleotides, proteins and alkaloids is another specific enzyme used as a marker of hepatic function. Elevation of the plasma level of this enzyme is also indicative or confirmatory of hepatic damage (Varley et al., 1980). There was a significant (0.05) decrease in the plasma ALP activities in all the groups treated for the first 3 weeks when compared with the normal and diabetic control groups (groups I and II) except for groups III. However, there was no significant (0.05) difference in the enzyme activities between the two control groups after 3 weeks of treatment. After 6 weeks groups IV and V had a significant decrease in the plasma ALP activities when compared with the normal and diabetic control groups (groups I and II). Group III had no significant (0.05) difference in the plasma ALP activities when compared with the two control groups. As the treatment progressed to the 9th week, there was no significant (0.05) difference in the plasma ALP activities between the normal and diabetic control groups. Only group V had a significant (0.05) decrease in the plasma ALP activities when compared with the normal control group. Groups III and IV, had no significant (0.05) difference in the plasma ALP activities when compared with the normal control group and diabetic control group (groups I and II).

 

From table 3, 4 above, it will be observed that the reference drug Glucophage (metformin, 1.4mg/kg) increased the plasma enzyme activities of ALP significantly (0.05) when compared with other groups throughout the period of treatment and this may be due to the adverse effect elicited by the drug on the liver.

 

Histological evaluation of the pancreas (fig.1-7) revealed a remarkable reduction in the islet cell mass of Langerhan of the streptozotocin induced diabetic rats throughout the period of treatment. As they were treated with metformin and its co-administration with reducdyn, the islet cells were repopulated and preserved throughout the period of treatment. Evaluation of the hepatic tissue after 3, 6 and 9 weeks of treatment revealed normal histology, mild inflammation and periportal inflammation of the normal control rat liver after 3, 6 and weeks of treatment respectively. The liver of the diabetic control rats was seen to have mild inflammation and tissue fragmentation after 3, 6 and 9 weeks of treatment. The administration of metformin alone induced periportal inflammation after 3 and 6 weeks and mild inflammation after the 9th week of treatment. Mild inflammatory change was observed on the liver of those treated with the combination of metformin and different doses of reducdyn throughout the period of treatment except for the 9th week where mild periportal and intraparenchymal inflammation was observed on the liver of those treated with metformin and the higher dose of redudyn (0.5mg/kg).

 

REFERENCES:

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7.        O'Brien, R. M. and Granner, D. K.(1991). Regulation of gene expression by insulin. Biochem J278: 609-619.

8.        Shimomura, I., Matsuda, M., Hammer, R. E., Bashmakov, Y., Brown, M. S. and Goldstein, J. L.(2000). Decreased IRS-2 and increased SREBP-1c lead to mixed insulin resistance and sensitivity in livers of lipodystrophic and ab/ab mice. Mol Cell6 : 77-86.

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Received on 25.08.2013

Modified on 10.09.2013

Accepted on 15.09.2013

© A&V Publication all right reserved

Research Journal of Pharmacognosy and Phytochemistry. 5(5): September – October 2013, 224-235