Experimental Study On The Effect Of Oblique Acupuncture On CaN / NFAT1 Signaling Pathway In Skeletal Muscle Of Contusion Model Rabbits

Objective:To observe the efficacy of acupuncture on behavior, histomorphlolgy, and the protein expression of CaN and NFAT1 of rabitts soft tissue after acute contusion of skeletal. To discuss the repair mechanism induced by acupuncture after skeletal muscle injury.Methods:30 rabitts were randomized into 5 group:blank group(n=6), model group (n=6), acupuncture treatment group (n=6), CsA group (n=6), acupuncture combined with CsA group (n=6). Except the blank group, the rest of the group use the acute skeletal muscle model of blunt trauma caused by the hit of a weight in free fall. Acupuncture group was intervened by filiform needle obliquely puncturing gastrocnemius muscle after modeling 24 hours. CsA group was intervened by CsA intraperitoneal injection of 9.25 mg/kg in the twice a day three days before modeling. Acupuncture combined with CsA group was intervened by acupuncture treatment combined with CsA: intraperitoneal injection. Model group received no intervention. Measure the weight of rabbits, gastrocnemius muscle swelling at the time before modeling and 24h,48h and 72h after modeling. The injured part of gastrocnemius muscle was withdrawn after 3 day of injury in each group, and then observing the change of muscle fibers, muscle cells and ultrastructure under the electron microscope after HE staining of the gastrocnemius every group. Using Western blot detect CaN and NFAT1 expression.Results:1. Oblique insertion can improve the swelling degree of the injured skeletal muscle rabbits, each rabbits’average weight at the time before modeling and 24h,48h and 72h after modeling have no statistical difference.;Compared with the blank group, the rest of the group of calf muscles swelling degrees were significantly increased. Compared with model group, CsA group, acupuncture group and the acupuncture combined wth CsA group after modeling reduced the degree of swelling of the gastrocnemius muscle. The acupuncture group relieve swelling degree is the most obvious.2. From the perspective of morphological observation, Oblique insertion has specific recovery effects on skeletal muscle tissue after acute contusion. Under light microscopic observation:Muscle cells of model group were edematous and irregular with small clearance, blurred boundaries and cytoplasm leakage. Inflammatory cells infiltration can be observed among muscle cells. The result of the CsA group indicated that muscle cells swell which far less than model group, inflammatory cell infiltration are less than model group appears a small cavity; The result of the acupuncture group indicated that muscle fibers arranged relatively continuous, the inflammatory cells infiltration is more obvious than model group, appears appregate disturbution. muscle cells arrangement is better overall. The result of the acupuncture combined with CsA group showed that muscle fibers arranged irregularly, the capillaries are visible, part of the muscle fibers dissolved. There is a small amount of cavitation and accompanied by a small amount of inflammatory cells infiltration. All the intevention groups have different degree to promote injured ultrastructural recovery. As for the improvement of ultrastructure injured tissue, the acupuncture group is most obvious.3. The result of CaN and NFAT1 expression tested by Western blot showed that:Compared with the blank group the rest of the group CaN protein were significantly increased (p<0.05); Compared to model group, CaN protein expression of the acupuncture group significantly increased (p<0.05); Compared to CsA group, CaN expression of acupuncture group and acupuncture combined CsA group were significantly higher (p<0.05). Compared with the blank group, NFAT protein expression of model group and acupuncture group were significantly higher (p<0.05 or p<0.01); Compared with model group, NFAT expression of acupuncture group was significantly increased (p<0.05), while CsA group and acupuncture combined CsA group had no statistical significance (p>0.05).Conclusion:1、Oblique insertion can effectively promote the recovery of skeletal muscle after the structural damage. CsA and acupuncture combined with CsA intervention both have a certain role in promoting recovery of skeletal muscle structure. CsA can affect on skeletal muscle repair after injury when used accompanied by acupuncture, it could be associated with CsA’ inhibiting function to the inflammatory response.2、CaN/NFAT1 signaling pathways plays an important role in the process of skeletal muscle regeneration repair after injury, acupuncture obliquely can activate the CaN/NFAT1 signaling pathway, thus promote regenerative repair of skeletal muscle damage. In addition, according to the results of this experiment, the function of oblique acupuncture for regenerative repair of skeletal muscle injury effect may be achieved by multiple pathways.