TSG (2,3,5,4'-Tetrahydroxystilbene-2-O- β -D-glucoside) from the Chinese Herb Polygonum multiflorum Increases Life Span and Stress Resistance of Caenorhabditis elegans

Abstract

2,3,5,4'-Tetrahydroxystilbene-2-O-β-D-glucoside (TSG) was isolated from Polygonum multiflorum, a plant which is traditionally used as an anti-ageing drug. We have analysed ageing-related effects of TSG in the model organism C. elegans in comparison to resveratrol. TSG exerted a high antioxidative capacity both in a cell-free assay and in the nematode. The antioxidative capacity was even higher compared to resveratrol. Presumably due to its antioxidative effects, treatment with TSG decreased the juglone-mediated induction of the antioxidative enzyme SOD-3; the induction of the GST-4 by juglone was diminished slightly. TSG increased the resistance of C. elegans against lethal thermal stress more prominently than resveratrol (50 μM TSG increased mean survival by 22.2%). The level of the ageing pigment lipofuscin was decreased after incubation with the compound. TSG prolongs the mean, median, and maximum adult life span of C. elegans by 23.5%, 29.4%, and 7.2%, respectively, comparable to the effects of resveratrol. TSG-mediated extension of life span was not abolished in a DAF-16 loss-of-function mutant strain showing that this ageing-related transcription factor is not involved in the effects of TSG. Our data show that TSG possesses a potent antioxidative capacity, enhances the stress resistance, and increases the life span of the nematode C. elegans.

Figures

Figure 1

Radical scavenging properties (TEAC assay) of TSG. Chemical structures of resveratrol, quercetin, and TSG are shown in (a); the antioxidative capacity of the substances measured by the TEAC assay is shown in (b). The decolourisation of the radical-solution was detected spectrophotometrically at 734 nm (mean values ± SD, n = 3, *P < 0.05 versus control value, Student's t-test).

Figure 2

Modulation of ROS accumulation in C. elegans by TSG. Modulation of ROS accumulation in wild type nematodes at 37°C after incubation with TSG, quercetin, and resveratrol (50, 100 μM). The fluorescence intensity of DCF (rfu) was used as a marker for intracellular ROS; mean ± SEM, 5–7 independent experiments; at least 80 animals per concentration were analysed; *P < 0.05 versus DMSO value, one-way ANOVA with Dunnett's post hoc test.

Figure 3

Modulation of SOD-3 expression by TSG. The images show GFP fluorescence of the head and anterior part of the intestine of CF1553 transgenic nematodes pretreated with 100 μM of the compounds without (DMSO) or with oxidative stress (150 μM juglone; 3 h). Data represent mean values ± SEM, n = 3, *P < 0.05 versus corresponding DMSO value, one-way ANOVA with Dunnett's post hoc test. At least 10 animals per group and experiment were analysed.

Figure 4

Modulation of GST-4 expression by TSG. The images show GFP fluorescence of the head and the anterior part of the intestine of CL2166 transgenic nematodes pretreated with 100 μM of the compounds followed by an incubation under physiological conditions or under conditions of oxidative stress (150 μM Juglone; 3 h). Data represent mean values ± SEM, n = 3, *P < 0.05 versus corresponding DMSO value, one-way ANOVA with Dunnett's post hoc test. At least 10 animals per group and experiment were analysed.

Figure 5

Increased resistance against lethal heat-stress by treatment with TSG. TSG treatment increases the resistance against thermal stress at 50 μM and 100 μM (a). Resveratrol treatment (b) and quercetin treatment effectively increased the survival only at 100 μM. Survival curves represent the data of 4 independent experiments with a total of 48 animals per group (Kaplan-Meier survival analysis); corresponding data are summarised in Table 1.

Figure 6

Prolongation of life span by treatment with TSG. (a) Pretreatment with 100 μM of TSG, resveratrol, and quercetin reduced the accumulation of the ageing marker lipofuscin (mean values ± SEM, n = 3, *P < 0.05 versus control value, Student's t-test). 20 animals per group and experiment were analysed. (b) All compounds (100 μM) induced a prolongevity effect during the complete adult life (Kaplan-Meier survival analysis of three independent experiments with at least 30 animals per group/experiment; survival data are summarised in Table 2).

Figure 7

The TSG-mediated prolongation of life span is independent of DAF-16. Treatment with the compounds (100 μM) during the complete adult life induced a prolongevity effect in the daf-16 loss-of-function mutant strain CF1038 (Kaplan-Meier survival analysis of two independent experiments with at least 30 animals per group/experiment; survival data are summarised in Table 2).