Evaluation of In Vitro, In Vivo and In Silico Antidiabetic Potential of Ethylacetate Extract of Amaranthus hybridus Leaf
- Prof. G.O. AJAYI’s supervision, and Dr F.J FEMI-OLABISI
- MOUNTAIN TOP UNIVERSITY BIOCHEMISTRY 2023
- 159P
Diabetes Mellitus (DM) has become a global endemic with severe consequences such as cardiovascular diseases and liver damage. Amaranthus hybridus has been reported ethnobotanically to be used in the treatment of DM. It is therefore necessary to give scientific validation to the acclaimed antidiabetic effect of A.hybridus. This study evaluated the phytochemical analysis and antidiabetic potential of ethylacetate extract of Amaranthus hybridus leaf (EAHL), using STZinduced diabetic wistar rats (in vivo) and inhibition of alpha glucosidase and alpha amylase activity (in vitro). In silico analysis was also carried out. For in vivo, forty-two male wistar rats weighing 200g to 250g were assigned into 6 groups (A- F), n= 7. Group A, Control; B, STZ (DM); C, DM + 300mg/kg BW of metformin; DM + 150mg/kg BW of EAHL; DM + 300mg/kg BW of EAHL; F, 300mg/kg BW of EAHL for 14 days. DM was induced with 60mg/kg BW of single dose STZ intraperitoneally. Blood glucose was checked using glucometer before and on the 2nd, 7th and 14th day of treatment. On the 14th day, the rats were kept in diethylether fume chamber, blood sample ere taken from the heart after cervical dislocation to estimate antioxidant parameters (SOD, MDA and CAT), liver function parameters (ALP, ALT and AST), Lipid profile, insulin assay, histopathology of liver and pancreas. The phytochemical screening of EAHL shows the presence of flavonoids, alkaloids, glycosides, terpenes while tannins, saponins and phlobatannins were absent. GC-MS results revealed bicycle [4.2.0] octa-1,3,5-triene (79%) and 1-heptafluorobutyryloxy-3-phenylpr (16.54%) as major bioactive constituents. Blood glucose level on day 14 was significantly lower (P<0.05) in Group A, C and E, when compared with Group B. Insulin was higher (p<0.05) in Control), DMEAHL, and EAHL when compared with DM. SOD was higher (p<0.05) in Control, DMMET, DMEAHL and EAHL when compared with DM. CAT was also higher (p<0.05) in Control, DMMET, and EAHL when compared with Group B. MDA was significantly higher in DM when compared with control and xii EAHL. The activities of ALP, ALT and AST were significantly higher (p<0.05) in the DM groups when compared to control, however, treatment with 300mg/kg of EAHL ameliorate this condition. The administration of EAHL at 300mg/kg significantly reduced (p>0.05) the concentration of total cholesterol, triglyceride, low density lipoprotein when compared to the control group, and also increase high density lipoprotein. For alpha-glucosidase EAHL showed 40.87% inhibition at 20 µg/ml, and this increased progressively with the concentration, reaching up to 81.80% inhibition at 100 µg/ml. For comparison, the standard acarbose showed greater inhibition across all concentrations, with 51.83% inhibition at 20 µg/ml and 88.08% at 100 µg/ml. Similarly, for alphaamylase, EAHL showed inhibitory activity starting from 38.78% at 20 µg/ml, increasing to 75.76% at 100 µg/ml. Again, acarbose demonstrated higher inhibition at all concentrations. This suggests that the ethylacetate extract of Amaranthus hybridus leaf could potentially have antidiabetic properties, given that both alpha-glucosidase and alpha-amylase are key enzymes in carbohydrate digestion and their inhibition is a common strategy in the management of postprandial hyperglycemia in diabetes. In silico test showed ligands with molecular weight not up to 500, none of the bioactive compounds appear to have better binding affinities for the receptor than the native ligand, particularly when NOE and metal constraints. The H and E stained section of the pancreasof group E shows pancreatic acinar cells display moderately pleomorphic nuclei but no striking mitotic figures. The results demonstrated that ethylacetate extract of Amaranthus hybridus possessed antioxidant and antidiabetic activities.Keywords: Diabetes mellitus, Ethylacetate Amaranthus hybridus Leaf