Combination therapy with Hordeum vulgare, Elettaria cardamomum, and Cicer arietinum exhibited anti-diabetic potential through modulation of oxidative stress and proinflammatory cytokines

Abstract

Poly-herbal therapies for chronic diseases like diabetes mellitus (DM) have been practiced in south Asia for centuries. One of such therapies comprises of Hordeum vulgare, Elettaria cardamomum and Cicer arietinum that have shown encouraging therapeutic potential in the treatment of diabetes and obesity. Therefore, poly-herbal granules (PHGs) of this formula were developed and investigated for their anti-diabetic and anti-obesity potential in obese-diabetic rats. The developed PHGs were chemical characterized and the virtual molecular docking was performed by Discovery studio visualizer (DSV) software. For in-vivo experiment, obesity in rats was induced with high-fat high-sugar diet. After that, diabetes was induced by alloxan monohydrate 150 mg/kg i.p. injection. The diseased rats were treated with PHGs at 250, 500 and 750 mg/kg/day for four weeks. GC-MS analysis of PHGs demonstrated the presence of 1,3-Benzenedicarboxylic acid bis(2-ethylhexyl) ester and 1,2-Benzenedicarboxylic acid di-isooctyl ester and phenol, 2,4-bis(1,1-dimethylethyl). Molecular docking of these compounds demonstrated higher binding energies with receptor than metformin against α-amylase and α-glucosidase. PHGs exhibited a decline in body weight, HbA1c, hyperlipidemia, hyperglycemia, and insulin resistance in diseased rats. The histopathological examination revealed that PHGs improved the alloxan-induced damage to the pancreas. Furthermore, PHGs increased the SOD, CAT and GSH while and the decreased the level of MDA in the liver, kidney and pancreas of diseased rats. Additionally, the PHGs had significantly downregulated the TNF-α and NF-κB while upregulated the expression of NrF-2. The current study demonstrated that the PHGs exhibited anti-diabetic and anti-obesity potential through amelioration of oxidative stress, NF-κB, TNF-α, and NrF-2 due to the presence of different phytochemicals.

Keywords: Diabetes; Inflammation; Obesity; Oxidative stress; Poly-herbal therapy.

Fig. 1

The effect of Polyherbal granules on body weight, glucose tolerance, and insulin level in diabetic obese rats. (A) body weight changes (B) Oral glucose tolerance test, (C) Time versus Glucose Level, and (D) insulin level in obese-diabetic rats. Data were displayed as mean ± SD: (n = 6). Body weight and blood glucose were analyzed by two-way ANOVA whereas insulin level was analyzed by one-way ANOVA followed by Tukey's test. ****P < 0:0001, ***P < 0:001, and **P < 0:01, and *P < 0.05 showed significant differences in comparison with the normal control group while ####P < 0:0001, ###P < 0:001, ##P < 0.01 and #P < 0.05 in comparison to the DC group.

Fig. 2

The effect of Polyherbal granules on glycated hemoglobin and lipid profile in diabetic obese rats. (A) HbA1c (B) cholesterol (C) triglycerides (D) HDL (E) LDL, (F) VLDL (G) cholesterol-HDL ratio and (H) lipids in obese-diabetic rats. Data were displayed as mean ± SD: (n = 6) and analyzed by one-way ANOVA followed by Tukey's test. ****P < 0:0001 and *P < 0.05 showed significant difference in comparison with the NC group while ####P < 0:0001, ##P < 0.01 and #P < 0.05 in comparison to DC group and ^^^^ P < 0:0001 and ^^P < 0:01, ^P < 0.05 in comparison to SC group.

Fig. 3

The effect of Polyherbal granules on oxidative biomarkers of the liver in diabetic obese rats. (A) Superoxide dismutase (SOD) in liver (B) Catalase (CAT) in the liver (C) reduced glutathione (GSH) in the liver (D) Malondialdehyde (MDA) in the liver; (E) SOD in kidney (F) CAT in kidney (G) GSH in kidney (H) MDA in kidney and (I) SOD in pancreas (J) CAT in the pancreas (K) GSH in the pancreas (L) MDA in the pancreas of obese-diabetic rats. Data were shown as mean ± SD: (n = 6) and analyzed by one-way ANOVA followed by Tukey's test. ****P < 0:0001, ***P < 0:001, and **P < 0:01, and *P < 0.05 showed significant difference in comparison with the NC group while ####P < 0:0001, ###P < 0:001, ##P < 0.01 and #P < 0.05 in comparison to DC group and ^^^^ P < 0:0001, ^^^P < 0:001, and ^^P < 0:01, ^P < 0.05 in comparison to SC group.

Fig. 4

Effect of treatment with polyherbal granules on the histology of the pancreas in diabetic obese rats at 40X magnification. A; Normal Control B; Disease Control (Diabetic), C; Standard Control (Metformin) pancreas, D; G-1 (PHGs 250 mg/kg) shows normal morphology of pancreas, E; G-2 (PHGs 500 mg/kg), F; G-3 (PHGs 750 mg/kg) in which orange arrow indicates islet cells and blue arrow indicates exocrine pancreas.

Fig. 5

The effect of PHGs on pro-inflammatory mediator in diabetic obese rats. (A) Tumor necrosis factor alpha; TNF-α (B) Nuclear factor kappa-B; NF-кB and anti-inflammatory mediator (C) nuclear factor erythroid-2; NrF-2 in obese-diabetic rats. Data were displayed as mean ± SD: (n = 6) and analyzed by one-way ANOVA followed by Tukey's test. ****P < 0:0001, and **P < 0:01 showed significant difference in comparison with the normal control group while ####P < 0:0001 and ##P < 0.01 in comparison to DC group and ^^^^ P < 0:0001, ^^^P < 0:001, and ^P < 0.05 in comparison to SC group.

Fig. 6

The 2D and 3D binding interactions of the compounds within the active pocket of an α-Amylase enzyme.

Fig. 7

The 2D and 3D binding interactions of the compounds within the active pocket of an α-Glucosidase enzyme.