| Aim:Study of that whether the hydrogen-water can reduce the rat hippocampal oxidative stress injury results from intensive and exhaustion training and its possible mechanism which will provide a scientific basis for the new antioxidant supplements.Method:There were all together56male SD rats aged of3months old with180-220g weight and were divided into7groups randomly which were defined as Group A (rats with exhaustion training and hydrogen-water), Group B (rats with exhaustion training and VE), Group C (rats with exhaustion training), Group D (rats with intensive training and hydrogen-water), Group E (rats with intensive training and VE), Group F (rats with intensive training) and Group J (control group). All these rats were fed by national standard rodents dry feed. After1week adaptive training, Group A, B, C, D, E and F were trained for4weeks. For Group A, B, C, the rats first received3weeks intensive training and then got1week exhaustion training while the Group D, E, F only underwent4weeks exhaustion training. The intensive training program included speed of15m/min in the first5min, speed of20m/min in the next5min and speed of25m/min in the last50min. For the exhaustion training plan, the speeds of Group A, B and C were15m/min in5min,20m/min in5min,25m/min in50min,30m/min in5min and25m/min in50min, however, the speeds of Group D, E and F were15m/min in5min,20m/min in5min and25m/min in50min.30min before the training, Group A, D were given hydrogen-water4ml by gavage, Group B, E were given VE4ml (20mg) by gavage, and Group B, E were given pure water4ml by gavage. The drawing materials time was chosen as24h after the last week training, in which time the rat hippocampus tissues were taken out and histologic sections were prepared in order to observe the changes of ultrastructures in the hippocampus. After that, the Hb, SDH and CK in blood were studied, the antioxidative indexes consisted of MDA, SOD and T-AOC are studied and the expresstion of autophagy proteins (mTOR, pmTOR and LC3B) was measured using Western-Blot.Results:1Compared with control group, Hb content, serum SDH and CK activity in the intensive training rats plasma rise sharply (p<0.01). Compared with intensive training group, plasma Hb content, serum SDH and CK levels are low in the group with VE supplement,but no significant;Compared with intensive training group, the group with hydrogen water supplement, plasma Hb content, serum CK SDH and was obviously lower (P<0.05);and, Compared intensive training with VE supplement group, the intensive training group with hydrogen water supplement group, plasma Hb content, serum SDH decr3ease, but no significant change, serum CK content significantly reduced (p<0.01).2Compared with control group, Hb content, serum SDH and CK activity in the exhaustion training rats plasma rise sharply (p<0.01). Compared with exhaustion training group,exhaustion training with VE supplement group, plasma Hb content, serum SDH and CK levels are low, but no significant; Compared with exhaustion training group, exhaustion training with hydrogen water supplement group, plasma Hb content, serum CK SDH and was obviously lower (p<0.05); and, Compared exhaustion training with VE supplement group, the exhaustion training group with hydrogen water supplement, serum CK and SDH decrease, but no significant change, plasma Hb content significantly reduced (p<0.01).3Compared with control group, the exhaustion time of exhaustion training with hydrogen water supplement group is much longer (p<0.05); Compared exhaustion training with VE supplement group, the exhaustion training group with hydrogen water supplement, rats, exhaustion time increases, but no significant differences.4From the SEM images, it can be seen that, in the normal rat hippocampus tissues, the structure of cell nucleus is normal, the graphs of nuclear pore complex and organelles included of endoplasmic reticulum, Golgi complex and mitochondria are clear, the chromatin is well distributed and autophagic vacuoles are inexistent. For the intensive training rats, the hippocampal neuron, endoplasmic reticulum, and mitochondria are swelling. And for the intensive training rats with VE supplement, the level of swelling is slightly lowered. Compared with the groups discussed above, for the groups with hydrogen-water supplement, the swelling of the hippocampal neuron, endoplasmic reticulum, and mitochondria are sharply reduced and the ultrastructures head towards normality. This phenomenon indicates that hydrogen-water can reduce the damage results from the intensive training and its effect is better than VE. 5For the exhaustion training rats, the hippocampal neuron and endoplasmic reticulum are swelling seriously, the cell permeability changes, the checked mitochondria cannot be seen clearly, the quantities of organelles decreases and bright areas appear in the cell nucleus. For the exhaustion training rats with VE supplement, the level of disorder in checked mitochondria is slightly lowered and for the groups with hydrogen-water supplement, the swelling of the hippocampal neuron, endoplasmic reticulum, and mitochondria are sharply reduced and the ultrastructures head towards normality. This reveals that hydrogen-water can also reduce the injury results from the exhaustion training and its effect is better than VE.6Compared with control group, the SOD activity, the content of MDA, and the level of T-AOC (p<0.05) in the intensive training rats hippocampus tissues rise sharply. The SOD activity and the level of T-AOC in the group with VE supplement increase more than that of intensive training rats, while the content of MDA is less than that of intensive training rats. The increases of SOD activity and the level of T-AOC (p<0.05) in the group with hydrogen-water supplement are much more and the the content of MDA decreases much less.7Compared with control group, the SOD activity, the content of MDA, and the level of T-AOC in the exhaustion training rats hippocampus tissues rise sharply (p<0.01). The SOD activity and the level of T-AOC (p<0.05) in the group with VE supplement increase more than that of exhaustion training rats, while the content of MDA is less than that of exhaustion training rats. The increases of SOD activity (p<0.05) and the level of T-AOC (p<0.01) in the group with hydrogen-water supplement are much more and the the content of MDA (p<0.05) decreases much less.8Western Blot results show that, compared with control group, the mTOR, PmTOR and LC3B express significantly of rat hippocampal of intensity training group are higher (p<0.01); Compared with intensive training group, expression of PmTOR, mTOR, LC3B protein are higher than those in the group with VE supplement,but no significant difference; Compared with intensive training group, the group with hydrogen water supplement, mTOR and PmTOR protein expression aer higher, but no significant differences, LC3B protein expression significantly is higher than intensive training group (p<0.05).9Compared with control group, the mTOR and LC3B express significantly of rat hippocampal of exhaustion training group are extremely higher (p<0.01), PmTOR express significantly of rat hippocampal of exhaustion training group are higher (p<0.05); Compared with exhaustion training group, exhaustion training with VE supplement group, mTOR, pmTOR, LC3B protein expression are increase, but no significant difference;Compared with exhaustion training group, exhaustion training with hydrogen water supplement group, mTOR, pmTOR, LC3B protein expression are increase (p<0.05); compared exhaustion training with VE supplement group, exhaustion training with hydrogen water supplement group, pmTOR mTOR and protein expression (p<0.05), LC3B protein expression have rising trend.Conclusion:1Added hydrogen water can effectively reduce damage of ultrastructure of organization structure in neurons in the hippocampus,which caused by the intensity training and exhaustion training,reduce the mitochondria swelling degree, protect the cell membrane and the mitochondrial form the structural integrity.2Added hydrogen water can dramatically reduce water force strength and dried rat plasma Hb training content, serum CK and SDH activity and improve the body anti-oxidant ability; Extend the rat exhaustion sports time, to promote the machine body movement ability.Hydrogen can be clearing parts of water free radicals, make the hippocampus organization the concentration of the free radicals is low. Hydrogen water can significantly reduce the MDA content in the hippocampus with intensity training group, which also can improve the SOD activity and total antioxidant ability in the hippocampus; Hydrogen water can significantly increase SOD, reduce the MDA content, extremely significant increase T-AOC level in the hippocampus with exhaustion training group, hydrogen-water can also reduce oxidative stress damage effectively.3Hydrogen water can greatly improve the protein expression of mTOR and pmTOR, reduce the protein expression of LC3B, in rat hippocampal of intensity and exhaustion training. Hydrogen water can protect the mitochondrial function, by the inhibition of cell of autophagy phenomenon... |
PDF Full Text Request