Skeletal muscle injuries can cause significant change in the ultrastructure and the metabolism of the skeletal muscle cells. Observation of the ultrastructure and measurements of the metabolism biomarkers such as total superoxide dismutase (T-SOD), malondialdehyde (MDA), and creatine kinase (CK) can be used to evaluate the degree of damage in human skeletal muscle injury. Rolling manipulation is the most popular myofascial release technique in Traditional Chinese Medicine. This study aimed to investigate the effects of intermittent pressure imitating rolling manipulation (IPIRM) of Traditional Chinese Medicine on ultrastructure and metabolism in the injured HSKMCs. Methods: In vitro techniques were used to culture HSKMCs, which were injured with high doses of dexamethasone sodium phosphate. Cells were divided into four groups-control normal group (CNG), control injured group (CIG), rolling manipulation group (RMG), and sine pressure group (SPG). RMG and SPG cells were cyclically exposed to 3.0 Kg (6.6 Pounds) of maximum force at a frequency of 2.0 Hz for 10 min in the Flexcell compression system for duration of 3 days continually. The cell ultrastructure, total superoxide dismutase (T-SOD) activity, malondialdehyde (MDA) content, and creatine kinase (CK) activity of the groups were assessed. Conclusion: These results suggest that the mechanical effects of rolling manipulation in TCM could not only improve the recovery of injured skeletal muscle cells by ameliorating organelles arrangement, reducing organelle swelling, and maintaining nuclear membrane integrity, but also ameliorate the functions of cellular metabolism by increasing T-SOD activity and decreasing MDA content and CK activity in injured skeletal muscle. Then the Hippo/Yap signal pathway was detected, and the proteins in each group were detected by Western Blot. The protein expression of upstream protein p-LATS1 and downstream protein p-Yap (Ser127) in each group was observed to explore the biomechanical mechanism of the method. The relative protein expression of p-LATS1 and p-Yap in (RMG) group was significantly higher than that in injured (CIG) group (P < 0.05). It was suggested that Hippo/Yap pathway was related to the stimulation of 3D human skeletal muscle cells, and the proliferation pathway of 3D human skeletal muscle cells could be opened by stimulation of three dimensional human skeletal muscle cells. It may be one of the biological mechanisms caused by the mechanical effects of manipulations in TCM.
Keywords: Flexcell FX-5000 compression system; Hippo/Yap pathway; Traditional Chinese medicine; cell ultrastructure; creatine kinase; human skeletal muscle cells; malondialdehyde; rolling manipulation; total superoxide dismutase.
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