Potential preventive and protective perspectives of different spice powders and their mixtures in rat model

Abstract

Background: The spices based dietary interventions are in lime light among the scientific community owing to their promising therapeutic perspective. The bioactive components in spices can be used to exert various health promoting functions in human body such as prompting weight loss, inhibit diet-induced obesity, hypercholesterolemia, hyperglycemia, allergies and various other maladies. In current study extraction and in vitro characterization of coriander seed (CS), black cumin seed (BCS) and fenugreek seed (FS) polyphenols was conducted for further development of dietary intervention against lipid and glycemia related abnormalities in experimental Sprague Dowley rats fed with control and different spice powder supplemented diets.

Methods: Purposely, extraction of Coriander (CS), Black cumin (BCS) and Fenugreek seeds (FS) were carried out by using water and aqueous methanol (70:30 v/v). Afterwards, the resultant extracts were thoroughly investigated for their antioxidant potential through different indices like TPC, TFC, FRAP and β Carotene Bleaching Assay and ABTS. Furthermore, HPLC quantification were also conducted with special reference to thymoquinone, disogenin, chlorogenic acid, caffeic acid and kaempferol alongside in vitro pancreatic lipase inhibitory activity estimation. Bio-evaluation trial was consisting of three modules i.e. study-I (normal diet), study-II (high cholesterol diet) and study-III (high sucrose diet). Furthermore, rats were sub-divided in five groups in each module on the basis of diet provision including T0 (control), T1 (Diet containing CS), T2 (Diet containing BCS), T3 (Diet containing FS) and T4 (Diet containing CSP + BCSP + FSP). At the beginning of trial, some rats were dissected to evaluate the baseline values whilst rest of the rats was killed at the termination (56th day). Feed and drink intakes were quantified on daily bases whereas, body weight was calculated weekly. Cholesterol level, serum low density lipoproteins (LDL), high density lipoproteins (HDL), triglycerides, glucose concentration and insulin level of collected sera was measured by standard procedures.

Results: The in vitro characterization showed better extraction of spices antioxidant through aqueous methanol as compared to water. Among the spices, Black cumin seed alone or in combination revealed highest antioxidant activity in T2 (BCS) followed by T4 (CS + BCS), T7 (CS + BCS + FS), T1 (CS), T6 (BCS + FS), T5 (CS + FS) and lowest in T3 (FS). Likewise, the HPLC characterization showed the presence of thymoquinone in BCS, Dosignienin FGS and chlorogenic acid, caffeic acid and kaempferol in the other treatments. Furthermore, all the treatments showed dose dependent inhibition in Pancreatic lipase activity and order of inhibition was BCS > CS + BCS > CS + BCS + FS > CS > BCS + FC > CS + FS > FS. The maximum feed intake, drink intake and weight gain was observed in T0 (control) trailed by T1, T2, T3 and T4 group in experimental study I, II and III, respectively. The resultant diet T4 enhanced the high density lipoprotein from T0 (58.58 ± 2.51) to 61.71 ± 1.62 (T4) in hypercholesterolemia rats whereas in hyperglycaemia rats the HDL was varied from 38.77 ± 1.2 to 40.02 ± 0.99 in T0 and T4, respectively. Similarly, T2 significantly lowered the low density lipoprotein from 62.53 ± 1.22 (T1) & 46.53 ± 0.99 to 54.88 ± 0.52 & 40.94 ± 1.99 (T2) in hypercholesteraemic and diabetic rats. Moreover, T4 treatment showed maximum reduction as 10.01 & 11.53% in respective studies.

Conclusions: The diet prepared from the different combination of spices has been proven effective against Oxidative stress related physiological malfunctioning.

Keywords: Black cumin seeds; Efficacy trial; Fenugreek seeds; Hyperglycemia; Lipid profile; Physical parameters.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1

a, b: Absolute values for different antioxidant indices carried out of water and methanolic extracts of different spices in alone and in combination. Two way ANOVA was applied to check the overall behaviour of the study parameter to elaborate the effect of treatments and solvent on different antioxidant indices . To evaluate the differences among the mean LSD test was (p ≤ 0. 05)

Fig. 2

a, b: HPLC chromatograms of Disogenin and Thymoquinone

Fig. 3

Pancreatic lipase inhibitory activity (%) of methanolic and water extracts of spices alone or in combination was evaluated on dose dependent manner by varying the concentration from 0 to 600 microgram/mL. Values are mean ± SEM (n = 03)

Fig. 4

a Feed intake by the experimental rats (g/rat/day) (A: fed with normal diet; B: fed with high cholesterol diet; C: fed with high sucrose diet). b Water intake by the experimental rats (mL/rat/day) (A: fed with normal diet; B: fed with high cholesterol diet; C: fed with high sucrose diet). c Weight gain by the experimental rats (g/rat) (A: fed with normal diet; B: fed with high cholesterol diet; C: fed with high sucrose diet)

Fig. 5

Effect of treatments on Absolute Average values for Hematological indices (Red blood cell indices and White blood cell indices) of all studies at the termination of trial (56th days). The experimental diets T0 Group rely on respective Experimental Diet without active ingredient), T1 (Group rely on respective Experimental Diet+ Coriander seed supplementation @ 1 g/Kg B.W) T2 (Group rely on respective Experimental Diet+ Black Cumin seed supplementation @ 1 g/Kg B.W), T3 (Group rely on respective Experimental Diet+ Fenugreek seed supplementation @ 1 g/Kg B.W) & T4 (Group rely on respective Experimental Diet+ Coriander Seed+ Black cumin seed + Fenugreek seed supplementation @ 0.333 + 0.333 + 0.333 g/Kg B.W) were given through entire study period. Values are mean ± SEM (n = 10) and level of significance were determined at (p ≤ 0. 05)