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Figure S1 HPLC assay of TCE. Two saponins were controls in TCE: 5β-furost-Δ25 (27) –en-1β,2β, 3β, 4β, 5β, 6β, 7α, 22ζ, 26-nonaol-26-O-β-D-glucopyranoside (saponin 1) and 5β-furost-Δ25(27)-en-1β, 2β, 3β, 4β, 5β, 7α, 22ζ, 26-octaol-6-one-26-O-β-D-glucopyranoside (saponin 2). HPLC analysis was applied on a Shimadzu 20A series HPLC system consisting of two pumps (LC-20A Solvent Delivery Unit), a column oven (CTO-10ASVP), a SPD detector (SPD-M20AV Photodiode Array Detector) and an LC solution Work Station. TCE was applied to himadzu VP-ODS column (5μm, 150 × 4.6 mm; Shimadzu, Japan) and detected at 203 nm. Column temperature was set up at 25°C and the flow rate was 1 ml/min. The mobile phase was acetonitrile – water two-phase gradient elution.

Figure S2 Effects of TCE on carbontetrachloride (CCl4) -induced hepatitis. Mice were given orally with TCE (75, 150, 300 mg/kg) or Bifendatatum (200 mg/kg) in PBS for five days, respectively. One hour after the final administration, Mice were intraperitoneally injected with CCl4 (0.12%, v/v, dissolved in olive oil, 10 ml/kg bodyweight) to induce acute liver injury, while the mice in the normal group were administered with equal volume of olive oil. The animals were sacrificed 24 h after CCl4 intoxication. Serum was obtained by bleeding for the measurement of alanine transaminase (ALT) and aspartate transaminase (AST) activities respectively. The data indicated the mean ± SEM of experimental animals (n = 10). One-way ANOVA revealed a significant difference at P < 0.01. *P < 0.05, **P < 0.01, vs Con A control; ##P < 0.01, vs normal (Dunnett's test).

Figure S3 Effects of TCE on picryl chloride (PCl)-induced contact hypersensitivity in mice. Mice were sensitized by painting 0.1 ml of 1% PCl in ethanol onto the shaved skin of their abdomen on day 0. Five days after sensitization, they were challenged on right ears with 30 μl 1% PCl in olive oil. At the time of challenge and 5 h later, TCE (75, 150, 300 mg/kg) were given orally twice. (A) Twenty hours after challenge, the ear swelling was evaluated. Each column represents the mean ± s.e.m of 12 mice. (B) Representative microphotographs showing ear histopathologic changes with hematoxylin–eosin staining (original magnification ×200). (C) Microscopic scores of ears (n = 12). One-way ANOVA revealed a significant difference at P < 0.01. *P < 0.05, **P < 0.01, vs Con A control; ##P < 0.01, vs normal (Dunnett's test).

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