دراسة مقارنة لبعض التأثيرات الفسيولوجية والنسيجية المرضية (الميتافورمين والإمباكليفلوزين) لدى الرجال وذكور الجرذان المصابة بالسكري من النوع الثان

A Comparative Study of Some Physiological and Histopathological Effects of (Metformin and Empagliflozin) on Male Humans and Rats Diabetics Type II

Summary

Glycemic control is still the most important aspect of type 2 diabetes mellitus (T2DM) treatment. Sodium-glucose cotransporter-2 (SGLT2) inhibitor, empagliflozin, and metformin therapy are effective and well tolerated in patients with type 2 diabetes mellitus. Metformin is the first of the several recommended treatment options. Since empagliflozin is poorly studied solely or in comparison with metformin, the current study aims to compare these drugs’ influence on glycemic levels individually in patients with type 2 diabetes mellitus (T2DM).

The study was a quantitative experimental investigation (cross-sectional study) that compared the influence of metformin and empagliflozin on the short-term glycemic level, the metformin group included 30 newly diagnosed type 2 diabetic patients who were treated with 1500mg of metformin which was divided 3 times daily and empagliflozin group includes 30 newly diagnosed type 2 diabetic patients who were treated with 10mg of Empagliflozin once time daily. A total of sixty male patients aged (30-70) years were enrolled in this study. This study was performed in the period from May 2022 to April 2023. All patients were already newly diagnosed as diabetic patients with type two Diabetes mellitus according to their medical history, American Diabetes Association (ADA), and World Health Organization (WHO) diagnostic criteria. They were informed about the medication’s dose, probable expected adverse effects, anticipated effects, and what they have to do in relation to their lifestyle and diabetic diary. In essence, patients were instructed to use their home glucose monitor (capillary blood) and to document their glucose readings four times per day. The timings were in accordance with the American Diabetic association and should include a fasting glucose reading and 1-2 hours post-meal readings. Blood and urine samples were analysed for basic and advanced investigations. A complete blood cell count (CBC), Lipid profile, Liver function test, Renal function tests, Thyroid Stimulating Hormone (TSH), and Glycosylated Hemoglobin A1C (HbA1c) test.
Additionally, measurement of the immunological parameter glutamic acid decarboxylase antibody (GAD65).
The results of human samples indicated that the metformin-treated group was shown a significant difference in the mean level of pre-post glucose test when compared to the empagliflozin-consuming group. The level of glucose was decreased significantly (P<0.001) in the group treated with metformin more than in the group treated with empagliflozin.

The study also included animal experiments, forty healthy, Swiss male albino rats of approximately (10–12 weeks old) with weights of (200 -225 g) were used in this research. The Induction of type 2 Diabetes Mellitus was performed through hyperlipidemia and insulin resistant model which was developed using the high fat and high sugar diet. High fat was prepared by mixing vegetarian ghee and oil in the ratio of 3:1 (v/v). it was given to the rats at a dose of 3ml per day. High sugar diet consisted of 25% sucrose which was orally fed to the rats for 8 weeks. Surviving rats after 8 weeks with a blood glucose concentration of 200mg/ml were considered HFHS-food-induced diabetic rats and used as models for further study. After the one-week adaption period, the rats were randomly divided into the Normal group:- the control healthy group (NG,n=10) and the HFHS-feeding rats group (n=30). At the end of 8 weeks, the HFHS-feeding rats were randomly divided into three groups (n=10), Group A rats with diabetes type 2 (DM2), Group B:- rates-DM2 treated with metformin 500mg/kg, Group C:- rats-DM2 treated with empagliflozin 10mg/kg. Both medications were administered to the rats-DM2 in experimental groups, and oral gavage for three weeks was applied. For all rats groups, the experimental rats were sacrificed and collected blood samples for assessment of hematological, immunological, and basic biochemical indicators in treatments, as well as, studying the histological changes in pancreas, liver and kidneys.
The results of the animal experiment showed that the RBCs number were decreased significantly (P<0.05) in rats-DM II group C that treated with empagliflozin compared to the groups A & B, the RBCs decreased by 18% in group C, and 8% in Group B respectively. The blood sugar level decreased significantly (P<0.05) in rats-DM II group B treated with metformin and in group C treated with empagliflozin compared to group A who was untreated, sugar levels decreased by 43% in Group B, and 35 % in group C respectively. The liver function test also showed a significant difference in the mean level of Alkaline Phosphatase (ALP) & Total Serum Bilirubin (TSB). Both markers were decreased significantly (P<0.001) in group B rates-DM2 treated with metformin and Group C rats-DM2 treated with empagliflozin. The mean level of both markers was decreased by about (44%).
Results also showed a significant decrease (P<0.05) in triglyceride concentration was observed in both treated diabetic rats’ groups (B&C) compared; the triglyceride level in the non-treated group (A) and the normal group was, however, reduced by 44% in the diabetic treated with metformin and 40% in the diabetic treated with empagliflozin. A significantly high(P<0.001) concentration of the mean level of immunity tests C-Reactive Protein (CRP) and Interlukein-6 (IL6) was observed in the untreated diabetic rats compared to the treated groups. Metformin treated group showed a low level of CRP and IL6 compared to the empagliflozin-treated group.

When pancreas, kidneys, and liver histological sections examined under a microscope, the DM2-rat pancreases which were shown increasing size islets Langerhans, hypertrophy cells of islets, and vascular congestion. The effects of the metformin in group B (DM2-rat) were clearly visible and included, normal size of islets Langerhans and no histological changes, While, the effect of empagliflozin in the pancreas of group C DM2-rat was shown still increasing size of islets Langerhans and mild congestion. The histological examination of DM2-rats liver indicated congestion with mild chronic inflammatory cell infiltration at the portal tract and lipid steatosis. The effect of metformin on the liver of DM2-rats shows the normal arrangement of hepatic cells with mild lipid steatosis, mild congestion, and no significant inflammation. Moreover, the effect of empagliflozin on the liver of group C included mild congestion and no inflammation. In addition, the DM2-rat kidney demonstrated congestion with mild atrophy of glomeruli and tubules with mild hyaline cast in some renal tubules with mild tubular epithelial cell degeneration. Among the histopathological consequences, using Metformin was shown to normal size of glomeruli mild congestion, mild hyaline cast in some renal tubules, and no inflammation or other pathology, but the effect of empagliflozin on the kidney of DM2-rat group C has included congestion, tubular epithelial cell degeneration and wide bowman space of glomerulus.