| Aging is an unavoidable natural law,and anti-aging is one of the important contents of medical research.Studies have shown that aging is closely related to body oxidation.The theory of free radical aging believes that the degenerative change in aging is caused by free radicals,especially reactive oxygen species?ROS?,which cause oxidative damage to cells and tissues,and with aging,free radicals keep accumulating.Excessive free radicals oxidize the body,damage cell membranes,affect the normal function of macromolecular substances such as proteins,and even cause DNA damage,resulting in mutations and inducing tumor formation.More and more researches show that antioxidant is an important step to prevent aging.Enzymes in free radicals or oxidants can break down tissues,affect metabolic functions,and cause various health problems.Antioxidant system is a self-protection mechanism formed by the body in order to protect it from excessive ROS.However,with the increase of age,the body will produce many reactive oxygen free radicals,resulting in redox imbalance.Hydrogen,as a reducing gas,can neutralize reactive oxygen species?free radicals?in blood and cells,and has strong anti-oxidant,anti-inflammatory and anti-apoptotic effects.Studies have shown that long-term drinking of hydrogen-rich water can delay aging,prevent depression,and protect the nervous system.Other studies have found that hydrogen can keep flowers and fruits and other plants fresh,and can delay the aging of rats'kidneys,gums and other parts.Bacterial fermentation is currently an important hydrogen production pathway.Clostridium butyricum?C.butyricum?is a common hydrogen-producing bacterium that can adjust the intestinal flora balance,promote the proliferation of beneficial intestinal flora,enhance immune function,and prevent tumorigenesis,but its mechanism of action is unclear.Therefore,this paper mainly explores the efficient preparation of hydrogen-rich water and the protective mechanism of C.butyricum on the body,in order to provide a reference for the application of hydrogen molecular medicine in clinical and daily life.1. The effect of ultrasonic deaeration treatment on the preparation of hydrogen-rich waterObjective:To detect the effects of ultrasonic waves on gas in water and the effects of ultrasonic water on the hydrogen production capacity of hydrogen rods,and to analyze the interaction between dissolved hydrogen,dissolved oxygen,redox potential,and p H value.Method:Distilled water was sonicated for 0,1,2,4,8,10,12,16 and 20 hours.The dissolved oxygen content,redox potential,and p H value were measured immediately.The hydrogen rod was placed in 12h ultrasonic water and distilled water.In a plastic bottle,the amount of dissolved hydrogen,dissolved oxygen,redox potential,and p H were measured immediately after 0,2,4,6,8,10,12,16,20 and 24 hours.Results:With the extension of the ultrasonic time,the oxygen content in the ultrasonic water gradually decreased,which was significantly lower than the oxygen content in distilled water at the same time point?p<0.01?.The amount of dissolved oxygen in the water reached the minimum value after 12 hours of ultrasonic treatment.The oxygen content in the water can gradually recover,and the change of the oxygen content conforms to the function law of y=A2+?A1-A2?/?1+?x/x0?^p??R2=0.94885,0.98391?.Compared with the ultrasonic water group and the distilled water group,the hydrogen production capacity is significantly different?p<0.01?,and the ultrasonic water group is better than the distilled water group.The amount of dissolved oxygen in hydrogen-rich water decreases with the increase of the amount of dissolved hydrogen,and the amount of dissolved hydrogen has a negative correlation with the amount of dissolved oxygen and the redox potential.Conclusion:Ultrasound can remove most of the oxygen in water.Ultrasonic water can significantly increase the hydrogen production rate of hydrogen rods in a short time and quickly prepare high-quality hydrogen-rich water.The study also produces references for increasing the hydrogen production rate of hydrogen rods,drinking hydrogen-rich water and clinical applications.2. Study on the mechanism of hydrogen-producing bacteria to delay the senescence of C.elegansObjective:To explore the hydrogen-producing ability of C.butyricum.Established a model of C.elegans cultivated with C.butyricum to clarify the effects of C.butyricum on the lifespan of different strains of C.elegans,and to investigate whether C.butyricum has beneficial effects on C.elegans through physiological,biochemical,and molecular indicators testing.And whether the beneficial effect of C.butyricum is through improving the antioxidant capacity of C.elegans.Methods:The hydrogen-producing ability of C.butyricum was tested.The N2 strains,sod-3 and sod-5 mutant strains were cultured with active and 60Co-inactivated C.butyricum.Compare and analyze lifespan and exercise ability?head swing,body flexion,throat pump vibration frequency?,respectively,detect changes in oxygen free radicals,SOD,CAT enzyme activity,and MDA content in different groups of C.elegans at 7 and15 days.The expressions of senescence and antioxidant-related genes were detected at 7and 15 days in C.elegans.Results:C.butyricum had strong hydrogen-producing ability and had normal activity in C.elegans.C.butyricum can prolong the lifespan of the C.elegans of N2 strains,sod-3and sod-5 mutant strains?p<0.05?,increase the frequency of head swing?p<0.01?,the number of bending of the body?p<0.01?and the frequency of pharyngeal pump vibration?p<0.05?.With the increase of culture time,ROS in C.elegans gradually increased,and ROS in C.elegans in the same age group were lower than those in the control group?p<0.05?.The SOD and CAT enzymes gradually decreased with the extension of the culture time.At the same age,the N2 and sod-3 strains of the C.elegans in C.butyricum group were higher than the control group?p<0.01?.The MDA content gradually increased with the extension of the culture time.In the same days,the N2 strain and sod-3 mutant strain of the C.elegans in C.butyricum group were lower than that in the control group?p<0.05?.At the transcription level,the age-1 gene expression increased with the increase of the culture time,and the C.butyricum group was lower than the control group?p<0.05?.With the increase of culture time,the expressions of sod-3,sod-5,daf-16,sir-1 and hsp-16.1 decreased,and the same age group in C.butyricum was higher than the control group?p<0.05?.Conclusion:C.butyricum has higher hydrogen production ability.C.butyricum can significantly reduce the body's ROS level,improve the body's own antioxidant capacity,and prolong the lifespan of C.elegans.3. Protective effect of C.butyricum on paraquat?PQ?induced oxidative damage in C.elegansObjective:To investigate the protective effect of C.butyricum on C.elegans under PQ oxidative damage,and to further clarify whether C.butyricum delays the senescence of C.elegans by improving its antioxidant capacity.Methods:C.butyricum and E.coli were used to cultivate N2 strain,sod-3,sod-5 mutant strains of C.elegans.Observe the changes of lifespan,body length,reproductive capacity and ROS.Results:The 0.5m M PQ exposure could significantly shorten the lifespan of C.elegans in N2 strains,sod-3 and sod-5 mutant strains,retarded growth and development,reduced egg production,and increased ROS accumulation in the body.C.butyricum prolongs its lifespan?p<0.05?,promotes growth and development,and restores part of its oviposition function?p<0.05?.The ROS accumulation in C.elegans increased with the increase of culture time.Under the same culture time,the ROS in C.butyricum group was significantly lower than that in PQ group?p<0.05?.Conclusion:The PQ exposure can cause C.elegans under oxidative stress.C.butyricum can improve the PQ damage and prolong C.elegans'lifespan.It may be because C.butyricum produces a large amount of hydrogen,which improves the antioxidant capacity of C.elegans.However,the specific protection mechanism needs further research. |
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