Anti-aging Effect Of Trilobatin On Caenorhabditis Elegans

Objective: To investigate the effect of trilobatinon prolongevity in Caenorhabditis elegans,and further to explore its potential mechanisms.Methods: All age-synchronized strains were cultivated for 2-3 generations on fresh NGM plates at 20 °C before used for experiments.The effects of different concentrations of TLB(62.5,125,250 ?M)on the lifespan of C.elegans were determined and further the optimal concentration was determined under normal culture conditions.Since 250 ?M TLB exhibited the best prolonged effect on lifespan,it was accepted as the optimal concentration in the following experiments.C.elegans were randomly divided into control group and TLB(250?M)group,and the effects of TLB on normal lifespan-associative markers in Caenorhabditis elegans were evaluated by measuring progeny production,body movement,lipofuscin and pharynx-pumping rate.Next,C.elegans were shifted onto plates with or without TLB(250?M)for 72 h at 20 °C.Thereafter,C.elegans were incubated for 2 h at 35 °C for thermo-tolerance assay or NGM plates containing paraquat at 20 °C for oxidative stress tolerance assay.The levels of ROS and MDA,the activities of GSH,SOD(SOD1,SOD2 and SOD3),CAT and GSH-Px in C.elegans were determined by ELISA assay.Furthermore,the fluorescence intensities of CF1139(DAF16::GFP),CL2166(GST4::GFP),KN259(SOD3::GFP),LD1008(SKN1::GFP)and TJ375(HSP16.2::GFP)were detected by fluorescence microscope.Finally,the effects of TLB on the lifespan of mutant C.elegans deficient in daf16,sir2.3,skn1,sod3,sod2 and gst4 were observed.Results: The results showed that TLB markedly prolonged the lifespan of wild-type C.elegans as evidenced by natural lifespan experiments;TLB observably heightened C.elegans resistance to thermal and oxidative stress tolerance;TLB attenuated ROS,MDA levels and increased antioxidant enzyme activities of SOD2,SOD3 and CAT,but not GSH,GSH-Px of C.elegans;TLB also significantly augmented the fluorescence intensity of DAF16::GFP,SKN1::GFP,SOD3::GFP,HSP16.2::GFP and GST4::GFP;Furthermore,longevity-enhancing effects of TLB were almost abolished in DAF16,Sir2.3,SKN1,SOD2,SOD3 and GST4 mutant C.elegans.Conclusion: Under this experimental condition,TLB effectively prolongs the lifespan of C.elegans,and its underlying mechanism is related to the regulation of SKN1/SIRT3/DAF16 signaling pathway and thereby increased the antioxidant capacity of C.elegans.