| Human lifespan is closely related to aging.After the organism matures,the senescence will accelerate with the growth of age and appear the physiological function irreversible decline.It has been found that hydrogen has a variety of biological activities such as scavenging free radicals,anti-oxidation,anti-inflammation and anti-metabolic disorders.In this study,the model organism,Caenorhabditis elegans,was used to construct an oxidative stress model with walnut quinone to explore the anti-aging activity of hydrogen and its mechanism.Specific research contents are as follows:(1)Firstly,wild-type Caenorhabditis elegans were taken as the research object,and the effects of hydrogen on the lifespan of N2 nematodes were investigated by growing at 20?.The results showed that the longest life span of nematode could reach 30 h after hydrogen intervention,and the longest life span of nematode could reach 22 h in the control group.The average and maximum life span of the experimental group were both higher than that of the control group.After 30 hours of hydrogen-rich water treatment,the lifespan of the nematodes was increased by 21.4%and 26.7%,respectively,under the treatment of low and high concentration hydrogen-rich water.Meanwhile,hydrogen could effectively inhibit the accumulation of lipofcin in nematodes.The level of lipofuscin in the body reflects the aging degree of the body.With the increase of life span,the accumulation of lipofuscin in the body gradually increases.The accumulation degree of lipofuscin is evaluated by measuring the optical density value of lipofuscin.The results showed that hydrogen could effectively inhibit the accumulation of lipofuscin in nematode,delay its senescence and prolong its lifespan.(2)The damage caused by neurodegenerative diseases is usually irreversible,and the decline of nervous system function worsens with the aging of the body.Alzheimer's disease,common in older people,is caused by oxygen free radicals produced by inflammation of damaged nerve cells.Cl4176 is A transgenic nematode that expresses human A? peptide in muscle tissue.A? deposited in muscle cells of nematode shows progressive paralysis phenotype.Through this model,the experiment confirmed that the hydrogen in c.elegans has against the toxic effects of amyloid protein,low concentration of hydrogen under the intervention of transgenic c.elegans CL4176 for A beta induced paralysis of no significant difference,high concentrations of hydrogen under the intervention of transgenic c.elegans CL4176 showed A beta induced paralysis delay,shows that high hydrogen can alleviate toxicity induced by beta-amyloid,which shows that the hydrogen with ease geriatrics synaptic loss in alzheimer's disease and subsequent neuron degeneration.Therefore,it is further proved that hydrogen has anti-aging activity.(3)In recent years,some studies have shown that some substances decrease their reproductive ability and physical growth and development ability while prolonging their life span.The results showed that hydrogen had no effect on the total number of eggs laid and body length while prolonging the lifespan of nematodes,indicating that hydrogen had no effect on reproductive capacity and physical growth.The locomotion ability of Caenorhabditis elegans was observed at the same time of longevity experiment.Muscle degeneration due to aging causes the nematode's motor ability to decline.After 15 days of culture in hydrogen environment,the head swing times of nematodes in hydrogen group were more than those in control group,and the difference was significant.The bending times /20 s of the nematode on the seventh day increased from 12 to 16 at most,and there was no significant difference in the later period.There was no significant change in the frequency /min of pharyngeal pump on the third day,but it increased from 150 to 180 on the seventh day,with significant difference.The results indicated that hydrogen had a positive effect on the locomotion ability of nematode,which made nematode have more anti-aging ability.(4)Studies have shown that the lifespan of Caenorhabditis elegans is related to its resistance to external stress.In this study,walnutone was used to simulate endogenous oxidative stress.In this experiment,the survival number of nematodes was detected,and it was found that the survival rate of the control group was 10%,the survival rate of the experimental group was 18% and 20%,respectively,after 13 h survival in the oxidative stress environment with the concentration of 300 ?mol/L walnutone.After treated with hydrogen-rich water for 24 h,the nematode resistance to oxidative damage induced by butrin was significantly improved.The results indicated that hydrogen could significantly improve the resistance of nematode under oxidative stress.The survival curves of insulin signaling pathway daf-2 and gonad signaling pathway GLP-1 were not significantly different from those of the control group under the condition of walnutone oxidative stress,and hydrogen could not improve the antioxidant capacity,which indicated that the antioxidant activity of hydrogen was dependent on insulin and gonad signaling pathway.The survival curves of CLK-1mutants in dietary restriction signaling pathway EAT-2 and mitochondrial signaling pathway were longer than those in the control group,and the antioxidant stress ability was enhanced,which preliminarily indicated that the antioxidant activity of hydrogen was not dependent on dietary restriction and mitochondrial signaling pathway.(5)The ability of nematodes to resist oxidative stress is related to the expression levels of SOD and GSH-Px enzymes,and increasing the activity of enzymes can enhance the ability of nematodes to resist oxidative stress in vivo and thus prolong their life.Therefore,we first studied the effects of hydrogen on the activities of SOD and GSH-Px in nematodes.The results showed that the activity of SOD enzyme in nematode was increased by 24.5% compared with the control group under high concentration of hydrogen-rich water.he preliminary results indicate that the mechanism of hydrogen oxidation may be the activation of SOD enzyme activity.In order to detect whether the increase of SOD activity was due to the change of hydrogen-induced enzyme gene expression,the expression of SOD-3 gene was further explored.The results showed that the higher the hydrogen concentration,the stronger the fluorescence intensity of SOD-3 ::GFP,and the more obvious the difference was.The results showed that hydrogen promoted the up-regulation of SOD-3 gene expression,thus enhanced the activity of antioxidant enzymes in nematode,enhanced the ability of nematode to resist oxidative stress,and prolonged its life span.(6)Daf-16 is a key gene in the IIS signaling pathway,and the effect of hydrogen on the longevity of mutant nematode Daf-16 was evaluated to be related to the IIS signaling pathway.The results showed that different concentration of hydrogen pair could not prolong the life of daf-16 nematode.Moreover,the expression level of daf-16 and daf-2 in nematodes was up-regulated,suggesting that the mechanism of hydrogen longevity extension may be related to the expression levels of key genes daf-16 and daf-2 in IIS signal transduction pathway.SOD-3 gene is a regulatory gene in the oxidative stress signaling pathway,and the expression of SOD-3 gene is up-regulated,indicating that hydrogen may activate the gene expression of daf-16,SOD-3 and daf-2 in the pathway,and thus has the function of regulating the expression of stress-resistant proteins... |
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