Dillenia indica L. attenuates diabetic nephropathy via inhibition of advanced glycation end products accumulation in STZ-nicotinamide induced diabetic rats

Abstract

The present study was aimed to evaluate advanced glycation end products (AGEs) inhibitory activity of alcohol and hydro-alcohol extract (DAE and DHE) of Dillenia indica L. (Family: Dilleniaceae) and its potential in treatment of diabetic nephropathy by targeting markers of oxidative stress. D. indica was evaluated for its in vitro inhibitory activity against formation of AGEs by using bovine serum albumin. Diabetes was induced in male Wistar rats by streptozotocin (65 mg/kg i.p.) 15 min after nicotinamide (230 mg/kg, i.p.) administration. Diabetic rats were treated with different doses of extracts (100, 200 and 400 mg/kg) to analyze their nephroprotective effect. Tissue antioxidant enzymes level was measured along with the formation of AGEs in kidney to assess the effect of D. indica in ameliorating oxidative stress. D. indica showed significant inhibition of AGEs formation in vitro. D. indica produced significant attenuation in the glycemic status, renal parameter, lipid profile and level of antioxidant enzymes proving efficacy in diabetic nephropathy. Moreover, D. indica produced significant reduction in the formation of AGEs in kidneys. The present study concludes that D. indica as a possible therapeutic agent against diabetic nephropathy.

Keywords: Advanced glycation end products; Betulinic acid; Hyperglycemia; Oxidative stress.

Graphical abstract

Fig. 1

HPLC chromatogram for DAE (A), DHE (B) standard Betulinic acid (C). Arrow indicates peak for betulinic acid.

Fig. 2

. a: Effect of DAE and DHE on body weight (g). Values are represented as Mean ± SEM (n = 6). Data was analyzed by using One-way ANOVA followed by Tukey's multiple test; a vs control; b vs Diabetic control; cD. indica extract 100 mg/kg; e vs Glimepride 10 mg/kg. p < 0.05. b: Effect of DAE and DHE on food intake (gm/rat/day). Values are represented as Mean ± SEM (n = 6). Data was analyzed by using One-way ANOVA followed by Tukey's multiple test; a vs control; b vs Diabetic control; cD. indica extract 100 mg/kg; e vs Glimepride 10 mg/kg. p < 0.05. c: Effect of DAE and DHE on water intake (ml/rat/day). Values are represented as Mean ± SEM (n = 6). Data was analyzed by using One-way ANOVA followed by Tukey's multiple test; a vs control; b vs Diabetic control; cD. indica extract 100 mg/kg; e vs Glimepride 10 mg/kg. p < 0.05.

Fig. 3

a: Effect of DAE and DHE on blood glucose level (mg/dl). Values are represented as Mean ± SEM (n = 6). Data was analyzed by using One-way ANOVA followed by Tukey's multiple test; a vs control; b vs Diabetic control; cD. indica extract 100 mg/kg; d vs D. indica extract 200 mg/kg, e vs Glimepride 10 mg/kg. p < 0.05. b: Effect of DAE and DHE on serum insulin level (μIU/ml). Values are represented as Mean ± SEM (n = 6). Data was analyzed by using One-way ANOVA followed by Tukey's multiple test; a vs control; b vs Diabetic control; cD. indica extract 100 mg/kg; d vs D. indica extract 200 mg/kg, e vs Glimepride 10 mg/kg. p < 0.05. c: Effect of DAE and DHE on AGEs in kidney (RFU/mg protein). Values are represented as Mean ± SEM (n = 6). Data was analyzed by using One-way ANOVA followed by Tukey's multiple test; a vs control; b vs Diabetic control; c vs D. indica extract 100 mg/kg; d vs D. indica extract 200 mg/kg; e vs Glimepride 10 mg/kg. p < 0.05.

Fig. 4

a: Histopathological changes in Kidney of normal and treated rats (H & E 10×). “a” shows the structure of glomerulus. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated. b: Histopathological changes in Liver of normal and treated rats (H & E 10×). “a” shows the structure of central vein in liver. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated. c: Histopathological changes in pancreatic islet of normal and treated rats (H & E 10×). “a” shows the structure of β-cells. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated.

Fig. 4

a: Histopathological changes in Kidney of normal and treated rats (H & E 10×). “a” shows the structure of glomerulus. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated. b: Histopathological changes in Liver of normal and treated rats (H & E 10×). “a” shows the structure of central vein in liver. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated. c: Histopathological changes in pancreatic islet of normal and treated rats (H & E 10×). “a” shows the structure of β-cells. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated.

Fig. 4

a: Histopathological changes in Kidney of normal and treated rats (H & E 10×). “a” shows the structure of glomerulus. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated. b: Histopathological changes in Liver of normal and treated rats (H & E 10×). “a” shows the structure of central vein in liver. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated. c: Histopathological changes in pancreatic islet of normal and treated rats (H & E 10×). “a” shows the structure of β-cells. (A) Normal, (B) diabetic nephropathy control, (C) Glimepride 10 mg/kg, (D) DAE 100 mg/kg treated, (E) DAE 200 mg/kg, (F) DAE 400 mg/kg treated, (G), DHE 100 mg/kg treated, (H) DHE 200 mg/kg treated and (I) DHE 400 mg/kg treated.