| BackgroundWith the development of global aging society,aging and age-related diseases are not only common issues in clinical medicine and scientific researches,but also have impacts on politics and economics.Commonly defined as gradually functional decline in the time-dependent manner,aging is an inevitable process of most living organisms.Characterized by a progressive loss of physiological integrity,it is always accompanied by the risks of many human age-related diseases such as neurodegenerative disorders,cardiovascular diseases,type 2 diabetes and various cancers.Therefore,how to delay the process of aging and eliminate the potential risk factors for the age-related diseases seem to be urgently required,which explains why humankind pays much attention to this attractive and fascinating subject since ancient times.Taken incomparable advantages into account,the roundworm C.eleganshas been considered to be an excellent system for studying molecular mechanisms in regulating animal aging and longevity.And the genetic pathways and biochemical processes that modulate aging and longevity are well conserved from budding yeast to the nematode worm Caenorhabditeselegans and mammals.Here,we will try to unveil the mechanisms ofthe gene dve-1 in aging and longevity in C.elegansso that it could give clues to the further studies for human aging and longevity.The homologous of DVE-1 in humans is SATB1/SATB2.,which has been reported to be associated with human digestive diseases,especially in colon cancer.In addition,SATB1 also contributes to the modulation of lifespan after DR treatment according to the previous data.C.elegans dve-1 plays an important role in the development of embryonic and participates in the signal pathway that responses to mitochondrial UPR.dve-1 has also been reported to be involved in aging and longevity,but the mechanism is still unknown.purposeIn this work we will try to find the mechanism of dve-1 in lifespan in C.elegans,which would give clues to unveil the function of human satb1/satb2 in aging and longevity and might also provide more implications to delay or prevent aging and age-dependent diseases.Methods1.Determine the lifespan and fecundity phenotypes afterdve-1RNAi treatment.And testing the lifespan and fecundity of dve-1 RNAi on dve-1::gfp to determine whether it could restore the phenotype of dve-1 RNAi.Testing the lifespan and fecundity of dve-1(tm4803).The dve-1(tm4803)and dve-1::gfp hybridization experiments were performed to obtain tm4803;dve-1 :: gfp homozygote,and its lifespan and fecundity were measured to detect whether the mutant dve-1(tm4803).2.daf-2;tm4803 lifespan and fecundity were not consistent with the level of a single mutant,but in the middle of two single mutant levels;daf-18;tm4803 had a lifespan below the two mutant levels;daf-16;tm4803 lifespan and fecundity at the level of two single mutants.3.hsp-6p::gfp,hsp-60p::gfp were treated with spg-7 RNAi and dve-1 & spg-7 RNAi respectively,and its fluorescence intensity was measured to detect the effect of dve-1 RNAi on spg-7-induced mitochondrial UPR response.Next tm4803;hsp-6p::gfp and tm4803;hsp-60p::gfp were obtained by hybridization experiments with hsp-6p::gfp/hsp-60p::gfp and mutant dve-1(tm4803),Respectively spg-7 RNAi on hsp-6p::gfp,tm4803;hsp-6p::gfp and hsp-60p::gfp,tm4803;hsp-60p::gfp.The effect of dve-1(tm4803)on spg-7-induced mitochondrial UPR response was analyzed.Finally co UPRehensive analysis of dve-1 on the regul ation of lifespan and mitochondrial UPR response.4.Extracting RNA from C.elegans N2 and dve-1(tm4803)of L4 for 2 days,Reversaled.act-3 was used as internal reference.The m RNA expression levels of sod-1,sod-2,sod-3 and ctl-1 and ctl-3 in N2 and dve-1(tm4803)were measured by RT-PCR.Results1.dve-1 RNAi will significantly reduce the lifespan and fecundity;overexpression of dve-1can not restore its level of lifespan and fecundity.dve-1(tm4803)will significantly reduce the lifespan and fecundity;Overexpression of dve-1 can restore its lifespan and fecundity levels.2.The lifespan and fecundity of daf-2;tm4803 were not consistent with the level of single mutant,but in the middle of two single mutant levels;The lifespan of daf-18;tm4803 at the below of two single mutants level;The lifespan and fecundity of daf-16;tm4803 level at the below of two single mutants.3.spg-7 RNAi on hsp-6p / hsp-60 p significantly enhanced the fluorescence intensity,dve-1& spg-7 RNAi weakened the fluorescence.However,the fluorescence intensity of spg-7 RNAi tm4803;hsp-6p::gfp?tm4803;hsp-60p::gfp was close to that of the control group.4.Compared with N2,the level of m RNA expression of sod-1,sod-2,sod-3 and ctl-1,ctl-3in dve-1(tm4803)decreased.Conclusions1.dve-1(tm4803)mutants and dve-1 RNAi worms show decreased lifespan compared with that in wild type worms.2.dve-1 may function in paralle with the classic IIS pathway.3.dve-1 modulation lifespan may not through its role in mitochondrial UPR.4.dve-1 involved in lifespan may be related to its function in oxidative stress. |
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