Effects And Investigation On Relative Mechanism Of Insulin-Like Growth Factor-1 On Cystationine-β-Synthase And Hydrogen Sulfide

MethodsAfter PC12 cells were treated with 20, 40 and 80 ng/ml IGF-1 for 24 h ,Western blot were used to analyze the proteins levels of CBS,fluorecence quantitative PCR were used to analyze the gene levels of CBS,sensitive sulphur electrode were used to analyze the gas levels of H2S .For inhibition studies, 25 or 50μmol/L PD 98059, a specific MEK inhibitor which is on the up stream of ERK, was administered 30 minutes prior to 80 ng/ml IGF-1 treatment,Western blot were used to analyze the proteins levels of ERK/MAPK,Then the above tests were repeated.Results(1)IGF-1 20ng/mL,40 ng/mL,80 ng/mL groups increased the gene expression of CBS in comparison with normal group(1±0.185 ;2.422±0.185;3.504±0.185;4.881±0.185,P≤0.05), with the increasing concentration of IGF-1, and this effect is more obvious,the change of protain of CBS was in accordance with gene. (2)IGF-1 20ng/mL,40 ng/mL,80 ng/mL groups increased the gene expression of H2S in comparison with normal group(3.380±0.282;6.408±0.282;11.468±0.282; 21.711±0.282, P均≤0.05), with the increasing concentration of IGF-1, and this effect is more obvious.(3)IGF-1 80ng/ml group increased the protain of pERK in comparison with normal group(1.074±0.0180;0.738±0.0180,P≤0.05),PD98059 groups(25,50 uM)reduced the protain of pERK in comparison with IGF-1 80ng/ml group(0.563±0.0180;0.260±0.0180;1.074±0.0180,P≤0.05), 50 uM group had more effect than 25 uM group ( 0.260±0.0180 ;0.563±0.0180,P≤0.05),and there was no obvious change in total ERK (F=0.105,P=0.955,P>0.05).(4)PD98059 groups ( 25,50 uM ) reduced the protain of CBS( 0.244±0.009 ; 0.149±0.009 ; 0.980±0.009 , P≤0.05 ) and H2S (4.979±0.341;2.614±0.341;24.342±0.341,P≤0.05)in comparison with IGF-1 80ng/ml group, 50 uM group had more effect than 25 uM group(0.149±0.009;0.244±0.009,P≤0.05),the same with H2S(2.614±0.341;4.979±0.341,P≤0.05).ConclusionIGF-1 increased the expression of CBS/H2S ,this effect may achieved ERK/MAPK signaling pathway .