| Objective:Calcium is a critical signal of intracellular injury and repair. By Observing how the eccentric exercise and acupuncture affect intracellular calcium concentration and calcium channel function, this study will investigate the trait of calcium signal which induces skeletal muscle injury and repair.Methods:Experiment one, rats were divided into 4 groups:3 weeks exereise(3E),3 weeks exereise and acupuncture(3EA),3 weeks acupuncture(3A) and 3 weeks control(3C).3E and 3EA were submitted to 3 weeks of eccentric exercise on a treadmill down a-16° decline.3EA and 3A were submitted acupuncture after exercise.Concentration of cAMP, SLN were measured by Elisa, protein expression of PDE4D3, FKBP12, PLB, PLB-ser16 were measured by western blot. Experiment two, Rat were divided into 4 groups:exereise(E), exereise and acupuncture(EA), acupuncture(A) and control(C), and were submitted one-time eccentric exercise. In 0-72 hours after exercise, the morphology of skeletal muscle cell were observed by transmission electron microscopy, intracellular calcium concentration were measured by x-ray EDS analysis, the measurement methods cAMP、SLN、PDE4D3、FKBP12、PLB、 PLB-serl6 is same to experiment one. Experiment three, rat muscle strips were perfused in vitro, and were made to exhausted by electrical stimulation. These muscle strips were divided into 4 groups, GC(control), GEA(acupuncture after electrical stimulation), PKA(perfused with PKA inhibitor H89)and NCX (perfused with NCX blocker CB-DMB). Cell morphology and calcium concentration were measured, methods is same to experiment one.Results:cAMP of 3E elevated(p<0.05), and there are rise trend in 3EA and 3A. SLN of 3E tend to rise, PLB and ser16/PLB of 3E elevated(p<0.05). PDE4D3 of 3E and FKBP12 of 3E and 3EA declined(p<0.05). The sarcomere of E became chaos in 6 hours after exercise, and the structure had not completely restored in 72h after exercise. The sarcomere of EA became chaos in 12 hours after exercise, and restored in 24h-48h. Intracellular Calcium of E rise in 0 hour after exercise (p<0.05). There was a rise trend in 12h-48h and Calcium rise in 72 hours(p<0.05). Calcium of EA tended to rise in 0 hour after exercise, and rise in 12 hours(p<0.01), from then it declined gradually. PDE4D3 of E rised in6-72 hours, and PDE4D3 of EA rised in 12-48 hours (p<0.05). FKBP12 of E rised in 0h-12h after exercise, and declined in 72 hours(p<0.05). FKBP12 of EA rised in 0h-48h. PLB of E rised in 6h-48h, PLB of EA rised in 0h-6h(p<0.05). Serl6/PLB of E declined in 0h-48h, serl6/PLB of EA rised in 12h-48h(p<0.05). SLN of EA rised in 6 hours after exercise(p<0.05). cAMP of E rised in 6h-12h and 48h-72h, cAMP of EA rised in Oh and 12h-48h(p<0.05). Level of injury in GC group was very serious, and the intracellular calcium concentration of GC was higher than GEA(p<0.05). Injury of GEA cell relieved greatly. Injury in PKA group was more serious than GEA, but less than GC. Intracellular calcium of PKA tended to rise. The Level of injury and intracellular calcium in NCX was roughly equal to GC group.Conclusion:Muscle cells appear calcium overload at instantly after eccentric exercise and it can induce damage. In 12h-72h after exercise, calcium oscillation signal of repair started. Acupuncture can decline calcium overload at instantly after exercise, and can promote tissue repair by strengthening calcium oscillations. Long term eccentric exercise can cause calcium-leak of RyR, and induces cell damage. Acupuncture can decline calcium overload and damage accumulation by PLB phosphorylation and maintaining PDE4D3 protein expression. At instantly after exercise, acupuncture can decline calcium overload mainly by activating NCX. Activation of cAMP-PKA by acupuncture also plays a part of calcium overload declining. |
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