Fig. 1

FT-IR spectrum of Galacan.

Fig. 2

HPLC chromatograms of monosaccharide determination. (a) Chromatogram of an acid-hydrolyzed solution of Galalcan and (b) mixture of standards (glucose:galactose = 1:1).

Fig. 3

HPLC chromatogram of a TSK gel GMPWXL column eluted with 0.05 M NaNO3 supplemented with 0.05% sodium azide. (a) Chromatogram of Galacan solution and (b) chromatogram of solvent (water).

Fig. 4

GC chromatograms of the Smith degradation product of Galacan and acetylized standards. (a–d) Acetylation product chromatograms of glycerol, erythritol, glucose and galactose. (e) Chromatogram of the Smith degradation product.

Fig. 5

The 1H NMR (a), 13C NMR (b), DEPT (c), 1H–1H COSY (d), HSQC (e), and HMBC (f) spectra of low-molecular-weight Galacan (LG) and possible structure in Galacan (g).

Fig. 6

Fluorescence photographs (a) and intensity (b) of labeled L. brevis in zebrafish untreated and treated with different concentrations of Galacan (n = 10). mCTRL: fish only pretreated with L. brevis; POS: pretreated fish were fed with live combined Bacillus subtilis and Enterococcus faecium granules; GAL: pretreated fish were fed with Galacan. Significant differences between the mCTRL group and the other groups were detected (** p < 0.01, *** p < 0.001).

Fig. 7

Antihyperglycemic effects of Galacan evaluated in zebrafish. (a) Blood glucose levels after the zebrafish model of diabetes was established by treatment with metformin or different concentrations of Galacan (n = 10). (b) Fluorescence intensity of pancreatic β cells in zebrafish after treatment with metformin or different concentrations of Galacan (n = 10). (c,d) Fluorescence observation of different zebrafish groups by fluorescence microscopy. CTRL: blank control; STZ: diabetes model control induced with streptozotocin; STZ + MTF: positive control of diabetes model treated with metformin; STZ + GAL: diabetes model treated with Galacan (62.5, 125, and 250 μg/mL). Significant differences between the STZ group and the other groups were detected (* p < 0.05 and *** p < 0.001).

Fig. 8

Antiaging effects of Galacan on zebrafish. (a,b) Fluorescence observation of different zebrafish groups by fluorescence microscopy. (c) Fluorescence intensity of senescence-associated β-galactosidase (SA-β-gal) after aging model zebrafish were treated with catalase and different concentrations of Galacan (n = 10). (d) Telomerase activity in cells of zebrafish embryos after treatment with catalase and Galacan (n = 3). CTRL: blank control; HP: aging model control induced with hydrogen peroxide; HP + CAT: positive control of aging model treated with catalase; HP + GAL: aging model treated with Galacan (125, 250, and 500 μg/mL). Significant differences between the HP group and the other groups were detected (* p < 0.05, ** p < 0.01, and *** p < 0.001).

Acknowledgments
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