| The effects and mechanisms of D-ribose supplementation on Sprague-Dawley rats' physiological function were systematically investigated by modem biological techniques, such as blood biochemical detection methods,HPLC,RIA,ELISA,cell culture,TEM and immunofluorescence cytochemistry.The effects of ribose on fluctuation of blood biochemical indicators and the metabolism of high-energy phosphates in heart and skeletal muscle immediately after swimming and 72h-recovery were investigated in current work.Besides, the effects of ribose supplementation on anti-fatigue and anti-hypoxia,and the protection of ribose on cardiac-myocytes damage induced by hypoxia/reoxygenation were also assayed. These results will provide theoretical basis for the application of ribose on medical and health care.The main results are as follows:1.The blood sugar concentration of ribose test group(RTG)was lower than that of swimming control group(SCG)immediately after swimming.The blood sugar concentration of high dose ribose group(HRTG)was lower than that of low dose ribose group(LRTG)and middle dose ribose group(MRTG).Thus,under such circumstances,the blood sugar concentration decreased with the ribose dose increasing.The blood sugar concentration of LRTG,MRTG and HRTG immediately after swimming was lower than that of just negative control.The blood sugar concentration was higher in SCG(p＜0.01)(5.28±1.93 vs. 3.24±0.57mmol/L),LRTG,MRTG(p＜0.05)(4.80±0.64 vs.3.24±0.57mmol/L)and HRTG compared with Normal Control Group(NCG)after 72h-recovery.The concentration of blood sugar after 72h-recovery was higher than that immediately after swimming in SCG, MRTG and HRTG.D-ribose reduced the concentration of blood sugar during swimming,increased the uptake and utilization of glucose and improved the exercise capacity of rats.A certain amount of blood glucose is the foundation of glycogenesis.Therefore the blood sugar raised by ribose after 72h-recovery could increase the uptake of glucose into liver,boost the synthesis of hepatic glycogen and the muscle glycogen,decrease the process of gluconeogenesis and enhanced the sugar reserve after swimming.2.Immediately after swimming,the serum lactic acid concentration was lower in RTG compared with RCG.The serum lactic acid concentration in HRTG was lower than that in LRTG and MRTG.The level of serum lactic acid in SCG was higher than that in NCG after 72h-recovery.The serum lactic acid concentration in RTG was lower compared with in NCG and SCG after 72h-recovery.Compared with in SCG,the serum lactic acid concentration in LRTG and MRTG was significantly lowered(p＜0.05)(3.85±1.03 vs.5.92±1.43;4.33±0.47 vs.5.92±1.43mmol/L,respectively),the level of serum lactic acid in HRTG was lowered markedly(p＜0.01)(3.63±1.44 vs.5.92±1.43mmol/L).D-ribose decreased the concentration of serum lactic acid during swimming,due to boosting the cycle of serum lactic acid and increasing the gluconeogenesis.That would provide more energy for skeletal muscle for swimming.D-ribose accelerated the clear of serum lactic acid after swimming,which was successful against the accumulation of serum lactic acid,and in favor of the recovery of physical function after swimming.3.The level of insulin was higher in RCG compared with NCG.The level of insulin was higher in SCG,LRTG,MRTG and HRTG compared with in NCG immediately after swimming.The level of insulin in HRTG increased significantly(p＜0.05)(86.45±29.34 vs. 33.91±6.64μIU/mL).The level of insulin in RTG was higher than that in SCG.Compared with in SCG,LRTG and MRTG,the level of insulin in HRTG significantly increased(p＜0.05) (86.45±29.34 vs.41.43±4.59;86.45±29.34 vs.45.45±12.16;86.45±29.34 vs. 46.28±11.53μIU/mL,respectively).The level of insulin increased with the ribose increasing.The level of insulin was higher in SCG,LRTG and MRTG after 72h-recovery compared with that immediately after swimming,the level of insulin in LRTG significantly increased(p＜0.05)(88.88±68.59 vs.45.45±12.16μIU/mL).The level of insulin in HRTG decreased significantly(p＜0.05)(52.48±11.13 vs.86.45±29.34μIU/mL)after 72h-recovery.After 72h-recovery the level of insulin in SCG,LRTG(p＜0.05)(88.88±68.59 vs. 33.91±6.64μIU/mL),MRTG and HRTG was higher than that in NCG.The level of insulin was increased by D-ribose.Insulin caused glucose reducing and increased the transfer of glucose into skeletal muscle.Then the skeletal muscle would obtain more energy and the ability of exercise was enhanced.The insulin could inhibit the glycogen decomposition and gluconeogenesis and enhance the synthesis of glycogen and the sugar reserve.All of the processes promoted the recovery of physical function after swimming.4.The blood sugar concentration increased and the level of insulin decreased in a dose-dependent manner during swimming.Blood sugar increased and insulin decreased with the ribose increasing from 100mg/100g·body·wt to 600mg/100g·body·wt.5.The levels of norepinephrine and epinephrine were higher in RCG compared with those in NCG and increased significantly in LRCG and HRCG(p＜0.05)(34.20±15.44 vs. 14.28±10.50;39.03±19.70 vs.14.28±10.50pg/mL,respectively).Immediately after swimming,the level of norepinephrine was higher in LRTG and HRTG than that in SCG,and the level of epinephrine was higher in RTG compared with that in SCG.D-ribose increased the release of norepinephrine and epinephrine.Norepinephrine and epinephrine boosted the glycogen decomposition and enhanced gluconeogenesis directly,thus caused the increasing of glucose.The release of insulin was inhibited by norepinephrine and epinephrine,thus the level of insulin decreased and the concentration of blood sugar increased indirectly.6.The functions of insulin,norepinephrine and epinephrine was are to maintain the stability of blood sugar level during exercise and keep the normal physiological function of heart and brain.7.The level of ATP after swimming was lower in SCG,LRTG,MRTG and HRTG in skeletal muscle than that in NCG immediately.It decreased significantly in SCG(p＜0.05) (68.82±5.18 vs.173.56±35.02μg/mL)and LRTG(p＜0.05)(63.28±19.16 vs. 173.56±35.02μg/mL)and significantly decreased in MRTG and HRTG(p＜0.01) (35.04±19.10 vs.173.56±35.02;37.35±31.44 vs.173.56±35.02μg/mL,respectively). Compared with in RCG,the level of ATP immediately after swimming reduced significantly in RTG(p＜0.01)(63.28±19.16 vs.182.81±66.79;35.04±19.10 vs.169.35±16.20; 37.35±31.44 vs.157.73±23.35μg/mL,respectively).The level of ATP increased significantly in LRTG(p＜0.05)(157.65±63.71 vs. 63.28±19.16μg/mL)and in MRTG and HRTG(p＜0.01)(146.60±63.18 vs.35.04±19.10; 142.73±68.44 vs.37.35±31.44μg/mL,respectively)in skeletal muscle after 72h-recovery.The level of ATP in RTG was not different from NCG and RCG.The level of ATP in SCG (18.28±8.61μg/mL)was lower(p＜0.01)than that in NCG(173.56±35.02μg/mL)after 72h-recovery.The synthesis rate of ATP in skeletal muscle was significantly enhanced and the exhausted ATP pool during swimming was completely recovered by D-ribose within 72h. D-ribose accelerated the recovery of energy.8.The level of TAN(AMP+ADP+ATP)immediately after swimming in skeletal muscle was lower in SCG(p＜0.05),LRTG(p＜0.05),MRTG(p＜0.01)and HRTG(p＜0.01) compared with in NCG The level of TAN was higher(p＜0.01)in LRTG,MRTG and HRTG compared with in SCG and not different from NCG after 72h-recovery.Compared with that immediately after swimming,the level of TAN was higher in LRTG(p＜0.05),MRTG(p＜0.01)and HRTG(p＜0.01)after 72h-recovery.The level of TAN was significantly increased within 72h by ribose due to the high synthesis rate of ATP.9.Compared with NCG,the concentration of ATP was higher(p＜0.01)in heart in LRCG, MRCG and HRCG(28.75±3.81 vs.13.29±2.01;32.35±2.07 vs.13.29±2.01;34.74±4.56 vs. 13.29±2.01μg/mL,respectively).The level of ATP in heart immediately after swimming was higher(p＜0.01)in LRTG,MRTG and HRTG(32.36±4.48 vs.21.24±6.20;40.26±4.18 vs. 21.24±6.20;36.12±2.42 vs.21.24±6.20μg/mL,respectively)than that in SCG.D-ribose significantly enhanced the of ATP concentration heart,so that ensured the energy supply to heart and kept the normal state of heart during swimming.10.The results showed that the synthesis rate of ATP in heart and skeletal muscle was accelerated significantly by low dose ribose(100mg/100g·bw·day),middle dose ribose (300mg/100g·bw·day)and high dose ribose(600mg/100g·bw·day).11.ATP consumed during swimming was replenished timely.The energy consumption was reduced by ribose and the clearance of serum lactic acid was accelerated to keep the pH of blood.The level of insulin was raised during swimming by ribose to increase the gluconeogenesis and maintain the stability of blood glucose.Thus the normal physiological functions of heart and brain were maintained.The time of rats swimming was prolonged by 50.21%(115.54±51.72 vs.76.92±41.19min)with ribose supplementation.The result showed that ribose could have the ability of anti-fatigue.12.D-ribose converted to glucose through phosphate pentose pathway.The cycle of serum lactic acid was accelerated and the gluconeogenesis was enhanced by ribose to raise the concentration of glucose,thus the normal physiological function of heart and brain was maintained.The synthesis rate of ATP was accelerated by ribose and the consumption of energy was slowed down.Therefore,the survival time under hypoxia was significantly prolonged(p＜0.05),27.73±5.37min in RTG and 25.49±2.11min in CG.The accumulation oxygen consumption was increased with ribose supplementation.It showed that ribose has the ability of anti-hypoxia.13.The activity of SOD was enhanced and the concentration of MDA was decreased by ribose.After 3h-hypoxia/2h-reoxygenation,the activity of SOD was lower(p＜0.05)in H/R than that in NC(20.44±4.38 vs.57.73±8.33U/mL).The activity of SOD was higher(p＜0.05) in H/R+Rib1,H/R+Rib2 and H/R+Rib3 than that in H/R(50.20±5.30 vs.20.44±4.38; 43.56±7.33 vs.20.44±4.38;38.92±4.80 vs.20.44±4.38U/mL,respectively),and similar with NC.After 3h-hypoxia/2h-reoxygenation,the concentration of MDA was higher(p＜0.05)in H/R compared with in NC(29.32±3.23 vs.14.11±1.03nmol/mL).The concentration of MDA was lower(p＜0.05)in H/R+Rib1 and H/R+Rib2 than that in H/R(15.22±2.07 vs. 29.32±3.23;18.37±2.09 vs.29.32±3.23nmol/mL,respectively),and similar with NC.The results showed that ribose could eliminate the excess free radicals,inhibit lipid peroxidation of membrane and keep the integrated structure of cell membrane.14.D-ribose could generate NADPH,enhance the activity of oxidase,accelerate the elimination of free radical,increase the concentration of GSH,inhibit lipid peroxidation of cell membrane and keep the integrated structure of cell membrane.Ribose boosted the generation of PRPP directly,thus accelerated the recovery of purine nucleotides and ATP pool. The increased ATP could lighten the cell edema,and Ca2+overload intracellular induced by hypoxia/reoxygenation.Therefore,ribose could increase the viability of cultured cardiac-myocytes and reduce the release of LDH after hypoxia/reoxygenation.The viability of cardiac-myocytes in H/R,H/R+Rib1,H/R+Rib2 and H/R+Rib3 was 59.9%,88.7%,86.8% and 79.7%,respectively after 3h-hypoxia/2h-reoxygenation.The concentration of LDH was lower(p＜0.05)in H/R+Rib1 and H/R+Rib2 than that in H/R(52.62±5.13 vs.70.28±7.97; 61.83±7.14 vs.70.28±7.97U/L,respectively).The releasing of LDH was significantly reduced by middle and high dose ribose.The results showed that ribose played an important role in cell protection.
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