| Objective：To investigate the effect of massage on injured rabbit quadricepsfemoris and try to understand the possible molecular mechanism of massagein repairing of skeleton muscle injury. We analyzed pathologic andhistomorphological change of rabbit quadriceps femoris muscle,reestablishment of microcirculation, expression of Desmin and α-actin andremodeling of muscle cytoskeletal, the relationship of acetylcholinesterase(AchE) activity and motor endplate (MEP), the enzyme activity of SDH,ATPase related to energy metabolism by comparing that in the rabbitsquadriceps femoris muscle treated or non-treated by massage therapy.Methods：Injured quadriceps femoris muscle model was made artificially in33New Zealand rabbits, which were randomly divided into3groups:self-repairing group (including7-day (3rabbits),14-day,21-day injurysubgroup (6rabbits/subgroup)), massage group (including7-day,14-daysubgroup,6rabbits/subgroup) and normal rabbits (3rabbits) as control. Thequadriceps femoris injured models were established with self-made lead hitter. Massaging for the therapeutic animal began at the8th day after themodeling. The injured and adjacent muscle was massaged by RT-N2Intelligent Massage Device for15minute everyday at3000~3100rpm.Massage treatment was continued for7-day or14-day for massaging rabbits.The injured quadriceps femoris was accepted detection by conventional andmicrobubble ultrasound at the end of management. After ultrasonography atthe designated time point, the rabbits were undergone euthanasia. Thepathologic change for each group was detected by H&E staining. Theexpression ofα-Actin、Desmin in the injured quadriceps femoris waschecked by using IHC and Western Blot. The repairing of cytoskeletonstructure was evaluated according to the light and dark bands whichdisplayed by IHC staining ofα-Actin and Desmin on the muscle fibers. TheEnzymohistochemical Assay was employed to detect the activity of AchE,SDH and ATPase.5of slides were arbitrarily chosen in each group and10oftypical visual fields or100muscle fibers under microscope were selected tocount the MEPs, to analyze the average diameter and section area of injuredfiber or the average optical density (OD) value of Desmin, α-Actin, SDH,SDH and ATPase using Image-pro Plus6.0or NIS-Elements BR3.2software. One-Way ANOVA and LSD-T test were carried out to determinestatistically significant difference by using SPSS17.0Statistics Software.P<0.05was considered as significant.Results： All rabbits were survival through the experiment. The normalquadriceps femoris fibers were arrayed in integrity, had no anydegeneration, necrosis in the section. Comparing to the normal control,tissue necrosis, broken, atrophic, degenerated and disarranged muscle fiberwere displayed in the slides of SR rabbits. In contrast, degeneration andatrophy of the injured fibers were reduced, and new blood vessels werepresent in the tissue derived from massage rabbits. The non-homogenousand diffusive echo was detected in the injured area of quadriceps femoriswith significantly weak echo at the badly damaged tissue derived from theSR rabbits. Homogenous echo was gradually observed in injured musclesfrom M7to M14rabbits. Especially, capillary reformation andre-establishment of microcirculation were found around the injured parts ofquadriceps femoris of M group of rabbits. Contrasting to its control SRrabbits, more MEP and large area of MEP was detected in each group ofmassage rabbits. The activity of AchE, SDH and ATPase were more in themuscle of M group of rabbits than in the SR rabbits (P<0.01). In the injuredmuscle fibers of SR rabbits, the regular transverse striations as observed asin the normal tissue displayed by Desmin andα-Actin staining were almostdisappeared. However, the bright and dark brands on the muscle fiberswere gradually recovered with the massage therapy in M rabbits. Matchingwith these repairing stutas, upregulated α-actin and Desmin expression werenotable in M group of rabbits than in the SR rabbits as checked by IHC staining and Western Blot (P<0.05). Moreover, the expression of α-Actinand Desmin were stronger in M14group that that in M7group (P<0.01).The rehabilitation of cytoskeletonal structure and activity of ATPase hadtime-dependant effectiveness in the repairing tissue derived from massagerabbits.Conclusion：Traditional Chinese Massage facilitated injured muscle repairing,probably by promoting capillaries genesis and microcirculationreestablishing, up-regulating gene expression associated with cytoskeletonalprotein and increasing activity of aerobic metabolism-related enzymes. |
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