| Royal jelly(RJ)is a natural functional product which has kinds of pharmacological activities,such as immunomodulatory,antitumor,antioxidant and anti-aging.These pharmacological activities were contributed to protein components in RJ.Our teamwork found that 4%of the RJ is the optimal dose to extend the lifespan of workers,a group of water soluble RJ proteins(RJP60)obtained by precipitation contained the main component for worker bees longevity extension.In this study,the water soluble proteins RJP60 were further separated to study their longevity prolonging effect on workers and its aging related genes expression.The further separation,purification and mass spectrum identification of RJP60.In this paper,ammonium sulfate with different saturation were separately used to precipitate RJP60.The components seperated were named RJP50 and RJP70.The results identified by LC-MS/MS were RJP50 mainly contained MRJP1,MRJP3,MRJP2 and some glucose oxidase,MRJP9;RJP70 contained MRJP3、Arp1 and a little MRJP7,MRJP5.The effects of RJ protein components on workers lifespan.Worker bees(Apis mellifera)were fed with 50%sucrose solution as the basic food,two protein groups(equal quantity water soluble RJ protein to RJ)as supplementary to compare the effects of different protein components on the lifespan of workers.Grouped as follows,BSA(0.016 g BSA+4.984 g 50%sucrose solution);4%RJ(0.2 g RJ+4.8 g 50%sucrose solution);RJP60(0.016 g RJP60+4.984 g 50%sucrose solution);RJP50(0.016 g RJP50+4.984 g 50%sucrose solution);RJP70(0.016g RJP70+4.984 g 50%sucrose solution).The results showed that 4%RJ and the groups with RJ protein highly significantly improved the survival rate of workers than BSA(P<0.01).The effect of RJP50 on workers lifespan was highly significant better than 4%RJ,RJP60 and RJP70(P<0.01).The average life of RJP50 group was 70.74%higher than that of BSA group,which was 18.46%higher than the RJ group.Results demonstrated that RJP50 had significant effect on workers longevity extension.Real time-PCR(two reference genes)analysis the effect of RJ and separated fractions on aging related genes expression in head.RT-PCR was performed to determine expressions of Vg,AmILP1,AmILP2,and AmAKH genes in the head of the workers.Results showed that,the workers fed with other groups significantly down-regulated the relative expression of AmAKH compared with the BSA group.There was no significant difference in the relative expression of insulin-like peptide ILP1 and ILP2 among all these treatments.Vg gene expressed in the RJP50 group,the RJP60 group and the RJ group were significantly higher for about 8-11 times,but the RJP70 group was not significantly different compared with BSA group.The results showed that the royal jelly,RJP60 and RJP50 could significantly affect the expression of AKH and Vg genes in the head of workers to prolong its lifespan. |
