| Objective To observe the efficacy of acupuncture and cyclosporine A (CsA) on behavior, damage serum marker enzymes, histomorphology, insulin-like growth factor-1(IGF-1) mRNA and protein expression in the reparative process of rabbit soft tissue after acute contusion. To explore the mechanism of the reparative process of acute blunt muscle being effectively promoted with acupuncture.Methods30rabbits (weight2.0-2.5kg) were divided into5groups:normal control group, model group, acupuncture group, cyclosporine A (CsA) group and acupuncture combined cyclosporine A (CsA) group,6in each. The method of free falling weight hitting was used to make the model of acute soft tissue contusion by hitting20consecutive times. Obliquely puncturing Ashi point was used in acupuncture group, CsA injection in CsA group, obliquely puncturing Ashi point and CsA injection in acupuncture combined CsA group, non-intervention in model group, and no hitting or any medicine injection in normal control group. The injured part of gastrocnemius muscle was withdrawn after3days of injury in each group. The expression amount of mRNA and protein was detected with methods of Real-time PCR and Western blot.Results1.Behavioral observation:(1) Weight:at the same time window (before building model,24h after building the model,72h after building the model), no statistical difference of weight was observed among groups (P>0.05); in the same group, no statistical difference of weight was observed among time windows (P>0.05).(2) The perimeter of gastrocnemius:at the same time window (before building model,24h after building the model,72h after building the model), compared with the normal control group, the perimeter of gastrocnemius of each model group and intervention groups increased significantly (P<0.05); compared with the model group, the perimeter of gastrocnemius of each intervention group decreased significantly, but no statistical difference was observed(P>0.05);,no statistical difference of the perimeter of gastrocnemius was observed among intervention groups (P>0.05).(3) The hardness of gastrocnemius:before building the model, no statistical difference was observed among groups. At the time window (24h after building the model,48h after building the model), compared with the normal control group, the hardness of gastrocnemius of each model group and intervention group increased significantly (P<0.05); compared with the model group, the hardness of gastrocnemius of each intervention group decreased significantly, but no statistical difference was observed(P>0.05); no statistical difference of the hardness of gastrocnemius was observed among each intervention group (P>0.05). At the window time of72h after building the model, compared with the normal control group, the hardness of gastrocnemius of the model group, CsA group and acupuncture combined CsA group increased significantly (P＜0.05); Compared with the model group, the hardness of gastrocnemius of each intervention group increased significantly, but no statistical difference was found(P>0.05); compared with the acupuncture group, the hardness of the acupuncture combined CsA group increased significantly (P<0.05).2.72h afer building the model, the content of CK and LDH in serum:(1) Compared with the normal control group, the content of CK in serum of the model group, the CsA group and the acupuncture combined CsA group increased significantly (P<0.05); Compared with the model group, no statistical difference was observed among intervention groups(P>0.05); no statistical difference was observed among intervention groups(P>0.05).(2) Compared with the normal control group, no statistical difference in the content of LDH in serum was observed between the model group and intervention groups(P>0.05); compared with the acupuncture group, the content of LDH in serum of the CsA group increased significantly (P<0.05).3.The pathological changes of gastrocnemius:(1) Under light microscope, in the normal control group:the transverse section of muscle fibers were polygonal and at equality size. The longitudinal section of muscle fibers was strip tape and arranged parallel to each other. In the model group, the transverse section of muscle fibers was irregular polygonal, while some muscle fibers were oedema, and a large number of inflammatory cell infiltration were diffuse inflammatory cell infiltration. Compared with model group,in acupuncture group, the transverse section of muscle fibers was irregular polygonal, almost at equality size and arranged parallel to each other. The local inflammatory cell infiltration was more obvious. In CsA group, compared with acupuncture group, the transverse section of muscle fibers was irregular polygonal, while the swelling degree of muscle fibers decreased, and some muscle fibers ruptured. In the acupuncture combined CsA group, the transverse section of muscle fibers were irregular polygonal,muscle fibers edema obviously; the space among muscle cells decreased; the inflammatory cell infiltration existed.(2) Under transmission light microscope, in the natural group, sarcomeres and myofilaments arranged neatly, while sarcomeres were completed, Z-lines, and M-lines were clear and continuous; in the model group, some muscle fibers were disordered, A-zones and I-zones were indistinguishable. Z-lines and M-lines were indistinct. In the acupuncture group, muscle fibers were normal, sarcomeres, and myofilaments arranged neatly. In CsA group, sarcomeres were in different sizes, and myofilaments were disordered. Some muscle fibers were fractured, and Z-lines and M-lines were fractured and moved. In the acupuncture combined CsA group, some muscle fibers, Z-lines and M-lines were fractured.4.Compared with the normal control group, the expressions of mRNA and protein of each group decreased significantly (P<0.05); compared with the model group, the IGF-1mRNA expression of CsA group increased significantly (P>0.05), while IGF-1protein expression decreased significantly (P>0.05), and the IGF-1mRNA and protein expression of acupuncture group increased significantly (P<0.05); compared with the CsA group, the IGF-lmRNA and protein expression of acupuncture combined CsA group increased significantly (P<0.05).Conclusion:1.The modeling method can successfully copy acute closed skeletal gastrocnemius muscle injury model.2.The reparative process of acute blunt muscle can be promoted effectively with the interventions of acupuncture and CsA, acupuncture combined CsA.3.CsA may inhibit the inflammatory responses of the injury skeletal muscle, partially reduce the repairing effect of Acupuncture; the acupuncture combined CsA is more effective than CsA.4.Acupuncture can promote the reparative process of acute blunt muscle effectively and promote the expression of endogenous IGF-1mRNA and protein in the local acute injured skeletal muscle. |
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