Preparation Of Hydrogen-rich Water And Its Mechanism Of Life Extension Of C.elegans

Aging is a physiological phenomenon in which a tissue,cell,or organ changes over time.Life spanis the time from birth to death.As a tool to measure the aging process,life span is invaluable in analyzing genes that regulate aging.The terms"life span"and"aging"are often intertwined,but they are not equivalent.The theory of aging induced by free radicals suggests that aging is due to normal oxidative metabolism byproducts caused by the accumulation of molecular damage.With age,the spontaneous decline of the body function will lead to the repair effect,the anti-oxidant effect will be weakened,and the accumulation of free radicals will further increase the harm of injury.However,the presence of a certain concentration of antioxidants in the body will help to prolong the life span.Studies have found that many substances with antioxidant activity often have the effect of prolonging the life span of Caenorhabditis elegans?C.elegans?.Hydrogen molecules,as the single-reducing gas molecule with the simplest structure formula,can selectively neutralize hydroxyl radicals?·OH?and peroxynitrite anions?ONOO-?in vitro and show good antioxidant effects.However,the current research on hydrogen is focused on its effects on various diseases,and there are relatively few studies on the preparation of hydrogen molecules and their impact on life span.Therefore,this study mainly discusses the preparation of hydrogen molecules from the effects of different solute solutions on the hydrogen production rate of hydrogen rods,and studies the effects of hydrogen molecules on aging and life span of C.elegans.The aim is to provide reference for the rational application of hydrogen production in clinical and daily life,and to reveal the mechanism of hydrogen's role in delaying body aging.The main findings are as follows:1.Preparation of hydrogen rich solution and its influencing factorsObjective:To compare the hydrogen bar on the hydrogen production rate in different solute solutions,and the correlation of hydrogen and oxygen in clear solution,the relationship between hydrogen and oxygen reduction potential and the hydrogen dissolved in M9 solution escape velocity.Methods:The hydrogen containing magnesium rods were used to produce the hydrogen solutions through chemical reaction with water.The concrete can be divided into:NaCl,Na₂SO₃,Na₂SO₄,CH₃COOH and CH₃COONa solutions group,preparation respectively0%,0.2%,0.9%,1%,3%of the different concentration of the solution and 1×M9 buffer,and respectively in 0,2,4,6,8,10 h after the determine solution content of hydrogen,oxygen and oxidation reduction potential and pH value,and stop into a point in time after hydrogen content in the M9 buffer.Results:The higher the concentration of Na Cl was,the higher the hydrogen production rate was.The content of hydrogen and oxygen in the solution was significantly negative related?R²=0.9306?,and the higher the hydrogen content was,the lower the oxygen content was.With the increase of hydrogen content in solution,the redox potential decreased gradually,while the pH value increased gradually.The hydrogen production rate of hydrogen bars in different solute solutions is Na₂SO₃>NaCl>Na₂SO₄?p<0.05 or p<0.01?.In acetic acid solution,hydrogen production rate was significantly higher than that of sodium salt solution?p<0.05?,and the hydrogen content increased gradually with time.The oxygen concentration was significantly lower than that of sodium salt solution?p<0.05?,and the oxygen content decreased gradually over time.In the 1×M9 buffer solution,the concentration of dissolved hydrogen?DH?has gradually increased while there is no significant difference on the pH value with the increase of hydrogen production time.In the 1×M9 buffer solution,the hydrogen content with the extension of time first increased when prepared with the machine to input the hydrogen gas directly and then no longer raise,after then the hydrogen content is decreased with the extension of time and gradually decline.Conclusion:We demonstrated that the hydrogen rods have different hydrogen production capabilities in different solute solutions,and the oxygen content in the solution has a negative correlation with the hydrogen production capacity of hydrogen rods.The buffer solution can prevent the pH changes of the hydrogen in the hydrogen rods from increasing.2.Mechanism of hydrogen delaying the aging of C.elegansObjective:The effect of hydrogen on the life span of C.elegans in different strains was clarified.Through the deep research on the physiological indexes of nematodes and the genes related to aging,we further revealed and elucidated the mechanism of hydrogen delaying aging.Methods:Hydrogen interfered with C.elegans in different strains.The life span of C.elegans in hydrogen group and OP50 group was compared.The daily survival of nematode was calculated and the survival curve was made.The reproductive?oviposition?ability,activity?oscillating frequency?and the level of reactive oxygen species?ROS?were evaluated;detection of natural aging aging-longevity-related gene expression changes.Results:Hydrogen can significantly prolong the life span of the N2 and sod-5nematodes.There was no significant difference in the number of eggs laid between the hydrogen group and the control group,but the head swing frequency was significantly increased.With the increase of age,the level of ROS in nematodes continued to increase,and the levels of ROS in N2 and sod-5 nematodes in hydrogen group were lower than those in control group.The aging-related genes age-1 and let-363 in the hydrogen group decreased,and the expression of longevity-related gene ins-18 was increased.Conclusion:We showed that hydrogen prolonging the life span of C.elegans has nothing to do with its reproductive ability,and may be related to the level of ROS in vivo.3.The protective effect of hydrogen on the damage ofC.elegans induced by paraquat?PQ?Objective:To establish a PQ oxidative stress injury model,and to clarify the protective effect of hydrogen on the oxidative damage caused by C.elegans in PQ.To establish a model of nematode PQ oxidative stress injury,and to clarify the protective effect of hydrogen on oxidative damage caused by nematode under PQ stress,further revealing that hydrogen anti-aging plays an anti-oxidative stress role.Methods:Hydrogen intervenes with different concentrations of PQ?0.5mM and 5mM?prepared C.elegans oxidative damage model.The life span of C.elegans in hydrogen group and PQ group was compared and analyzed.The daily survival of nematode was calculated and the survival curve was made.The body length,adult rate,fecundity and changes of reactive oxygen species in vivo were compared between groups.Results:The life of nematode worm can be prolonged under the condition of 0.5mM PQ,and the life of C.elegans is close to normal.The length of N2 and sod-5 C.elegans in the PQ group were significantly smaller than that in the control group,while the C.elegans in the H₂ group were significantly lower than that in the control group.The adult and spawning numbers of N2 andsod-5C.eleganswere lower than that of the hydrogen group.The ROS levels in the N2and sod-5C.elegansin the hydrogen group were lower than that in the PQ group.Conclusion:We demonstrated that the hydrogen gas protects C.elegans by reducing the level of ROS in C.elegans,thereby prolonging life span of C.elegans.