| 1. ObjectiveThe experiment by observing the influence of the oblique to an acute eccentric exercise, to explore the mechanism of acupuncture promoting repair of skeletal muscle injury,by measuring the serum CK activity, MPO activity of skeletal muscle and the Ca2+ content of mitochondria and cytoplasm in the rat skeletal muscle injury model.2. MethodsExperimentⅠ: 96 male Wistar rats were randomly divided into 4 groups: control group(C, n=6), exercise group(E, n=30), exercise and acupuncture group(EA, n=30) and exercise, acupuncture and blocking agents group(EAI, n=30). Downhill running 90 min to establish acute eccentric exercise model by-16 degrees. The movement of the acupuncture group was treated by Ashi oblique movement of acupuncture intervention, blocking agent group were treated by acupuncture combined with Gd C l3 injection intervention, quiet group and exercise group without intervention. In 6 h, 24 h respectively after exercise, 48 h, 72 h, 120 h unit four biceps on experimental rats(mainly for the muscle of vastus internus and vastus medialis) damage parts materials, detection of Ca2+ content in skeletal muscle mitochondria; MPO, the content of serum CK detection in skeletal muscle tissue.ExperimentⅡ:24 male Wistar rats were randomly divided into 4 groups: control group(C, n=6), electrical stimulation group(E, n=6), electrical stimulation of acupuncture group(EA, n=6), electrical stimulation of acupuncture blocking agent group(EAI, n=6). By electrical stimulation of rat spintrapezius muscle to establish the model of damage excessive centrifugal movement. Exposure of the spintrapezius muscle in rats in vivo, elongated after electrical stimulation, muscle contraction(process is equivalent to the centrifugal motion). Electric stimulation using wave width 700 ms,8 V,100 Hz, 3 s each time, 50 times(150 s for each set); 5 min resting once, measuring a total of 10 sets.Control group without electrical stimulation. Electrical stimulation group electric stimulation; electric stimulation on after stimulation of acupuncture group(acupuncture needling the spintrapezius muscle, with a diameter of 0.25 mm filiform needle for 4 min); electrical stimulation of acupuncture blocking agent group in after stimulation of acupuncture. All groups were incubated in stimulating reagents and record the changes of muscular tension.Take in the body the spintrapezius muscle shooting Ca2+ fluorescence images of muscle tissue in vitro, removed for determination of spintrapezius muscle mitochondrial Ca2+.3. ResultsExperimentⅠ:3.1 The activity of serum CK in different groups: compared with C, E is increased significantly at 24 h, there is significant difference(P<0.001); 48 h has a very significant difference(P<0.01), 120 h to the quiet group; group EA and 24 h reached the peak. There was significant difference(P<0.05), 48 h to 120 h has been decreased to the level of quiet group; group EAI, 6 hours, there was a significant difference(P<0.01), 24 h, 48 h and 120 h had very significant difference(P<0.001), while 120 h decreased, but higher than that of E and EA gro up. Each time point, compared with EA, EAI and exercise group, 24 h EA and 48 h EA and there was significant difference(P<0.05), 6 h EAI and 48 h EAI and there was significant difference(P<0.05). EAI compared with EA, there were significant differences in 24 h(P<0.05), there was a significant difference in 6h and 120 h(P<0.01), there was a significant difference in 48 h and 72 h(P<0.001). EA and EAI group at each time point, the 24 h had significant difference(P<0.05), there are very significant difference(P<0.01).3.2 Different groups of rat femoral muscle cytosolic activity:compared with C, 6 h EAI, 6 h E, 24 h E, 24 h EAI and 72 h EAI have significant difference(P<0.05); 48 h EAI had very significant difference(P<0.01); 6 h EA had very significant difference(P<0.001). Each time point, the other group and the exercise group compared to 24 h EA and 72 h EAI have significant difference(P<0.05); 6 h EA and 48 h EAI had very significant difference(P<0.01). Each time point, EA compared with EAI, there was a significant difference in 48 h(P<0.01), there was significant difference in 72 h(P<0.05). The group returned to normal level in 120 h.3.3 Mitochondria Free Ca2+ in different groups of rat: compared with the control group, 24 h EA and 24 h EAI have significant difference(P<0.05); 24 h EA and 48 h EAI had very significant difference(P<0.01); 6 h EA had very significant difference(P<0.001). Each time point, the other group and the exercise group compared to the 48 h EAI, there was significant difference(P<0.05); 6 h EA had very significant difference(P<0.001). Each time point, EA compared with EAI, there was a significant difference in 6 h(P<0.01), there was significant difference in 48 h(P<0.05). Each group was first increased and then decreased and recovered to normal level at 120 h,but the peak value of E, EA and EAI were appeared in 24 h, 6 h and 48 h, and the peak value was EA > E > EAI.ExperimentⅡ:3.1 Calcium fluorescence image display: The muscle fiber fluorescence intensity of the control group(C) gray images is weak and unobvious, no fluorescence in the cytoplasm. The electrical stimulation group(E), muscle fibers and cytoplasm fluorescence is very strong, the visible region most of the muscle fibers has much strong fluorescence and scattered spots. Electrical stimulation of acupuncture group(EA), the majority of muscle fiber decreased fluorescence intensity and fluorescence, highlights a lot. The movement of acupuncture plus blockers(EAI), muscle, a muscle fiber strong fluorescence. The average gray value of fluorescence image display, free calcium concentration of rats in different groups in the trapezius muscle cytoplasm, compared with the control group, the E had a very significant difference(P<0.001); compared with E, EA and EAI had very significant difference(P<0.01).3.2 The concentration of free calcium in the spintrapezius muscle mitochondria, compared with C, EA had very significant difference(P<0.01); compared with E, EA has a very significant difference(P<0.01); compared with EA, EAI has a very significant difference(P<0.01).3.3 The spintrapezius muscle force: compared with control group, 90 min after electrical stimulation, E, EA,EAI had a very significant difference(P<0.01); 100 min EAI E, there was significant difference(P<0.05). 100 min, compared with E, EA had significant difference(P<0.05); compared with EA, EAI had significant difference(P<0.05).4 Conclusion4.1 Elevated cytosolic Ca induced by eccentric contractions of skeletal muscle, can also increase the serum CK activity and muscle MPO activity.4.2 Acupuncture can effectively reduce the intracellular Ca overload phenomenon, promote mitochondria to absorb Ca2+, and then induce inflammatory reaction, accelerate the tissue repair damage.4.3 Activation of stretch activated channels by acupuncture, induce mitochondrial calcium uptake and reduce intracellular Ca2+, and then induced a list of inflammatory reaction, thus play a role in the injury and repair. |
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