Abstract
Objective To evaluate the in vitro anti-diabetic effects of Bryonia dioica roots extracts,in-cluding water-acetone extracts and their ethyl acetate and butanol fractions,and chloroform-methanol extracts.
Methods The total phenolic,flavonoid,flavonol,and saponin contents in the Bryonia dioica root extracts(chloroform-methanol extracts,water-acetone extracts and their ethyl acetate and butanol fractions)were determined using colorimetric methods with Folin-Ciocalteu,aluminum trichloride,and vanillin reagents,respectively.The in vitro anti-diabetic activity was evaluated by measuring the half-maximal inhibitory concentration(IC50)values of these root extracts against α-amylase and α-glucosidase activities,evaluating their effects on α-amy-lase kinetics,quantifying the inhibition of bovine serum albumin(BSA)glycation using fluo-rometry to assess advanced glycation end products(AGE)production,and determining glu-cose uptake by isolated rat hemidiaphragm.Additionally,molecular docking analysis was conducted to investigate the binding affinity and interaction types between Bryonia dioica lig-ands(cucurbitacin B,bryogénin,vitexin,and isovitexin)and target enzymes,and a phyto-chemical-targets interaction network was constructed.
Results For α-amylase inhibition,ethyl acetate fraction demonstrated the most potent activi-ty(IC50=145.95 µg/mL),followed by chloroform-methanol extract(IC50=300.86 µg/mL).Water-acetone root extracts and their ethyl acetate and butanol fractions inhibited the α-glucosidase activity with IC50 values ranging from 562.88 to 583.90 µg/mL.Both ethyl acetate and butanol fractions strongly inhibited non-enzymatic BSA glycation(IC50=318.26 and 323.12 µg/mL,respectively).The incubation of isolated rat hemidiaphragms with the ethyl acetate fraction(5 mg/mL)significantly increased glucose uptake(35.16%;P<0.000 1),exceeding the effects of insulin(29.27%),chloroform-methanol extract(24.07%),and catechin(15.27%).Molecular docking revealed that cucurbitacin B exhibited the strongest docking scores against α-amylase(-16.4 kcal/mol),and α-glucosidase(-14.2 kcal/mol).Compared with other ligands,isovitexin formed the maximum number of hydrogen bonds with the α-amylase active site residues(Asp300,Asp197,and Glu233),α-glucosidase residues(Ser13,Arg44,Met86,Gly10,Asp39,and Tyr131)and other residues(Arg195,Trp59,His299,and Tyr62).Network analysis identified 36 overlapping targets between Bryonia dioica phyto-chemicals and type 2 diabetes mellitus-associated genes,with cucurbitacins and polyphenols interacting with α-amylase,α-glucosidase,and Glut4 translocation pathway targets.
Conclusion Bryonia dioica root extracts demonstrated promising in vitro anti-diabetic activi-ty through multiple mechanisms,including the inhibitory effect on digestive enzymes,pro-tein antiglycation potential,and enhancement of glucose uptake,suggesting their potential as a source for anti-diabetic drugs development.关键词
泻根/葡萄糖摄取/蛋白质糖基化/α-淀粉酶/α-葡萄糖苷酶/分子对接/相互作用网络Key words
Bryonia dioica/Glucose uptake/Protein glycation/α-Amylase/α-Glucosidase/Molecular docking/Interaction network
