| Objective: To research the mechanical signaling pathways of injured human skeletal muscle cells by multi-cellular mechanical simulation platform,and to explore the transduction pathways of rolling manipulation from mechanical signals to electrophysiological and chemical signals.Method: This research will consist of three parts.The first part is the simulation of the stress cell loading system of Flexcell cells.The Fx-5000 cell stress loading system is combined with the sputum curve of the old Chinese medicine doctor Juntao Yan to realize the curve of Fx-5000 cell stress loading system.Professor Yan’s curvilinear curve is consistent.The second part is the cell experiment: 1.Purchase frozen human primary skeletal muscle cells for resuscitation and culture.2.Performing a model of injured human skeletal muscle cells.3.Make a three-dimensional human skeletal muscle cell gel.4 cells were divided into group: normal group(N),injury group(CIG),rolling manipulation 0.301Mpa(RMGA)group,rolling manipulation 0.603Mpa(RMGB)group,rolling manipulation 0.938Mpa(RMGC)group,6 samples per group.5.The Fx-5000 cell stress loading system intervened in the A,B,and C groups,respectively.The parameters were: 0.301 MPa,0.603 MPa,and pressure 0.938 MPa,continuous intervention for 3 days,10 minutes a day.6.Collect samples from each group and measure the total superoxide dismutase activity,malondialdehyde value,and creatine kinase value of each sample,and compare the optimal pressure values by sputum method.7.Hippo/Yap signal pathway detection: intracellular proteins were collected for western blot detection of p-LATS1,p-MST1,p-Yap cell proliferation and apoptosis indicators.Results: 1.Using the Flexcell cell stress loading system combined with the traditional Chinese medicine method,the sputum curve was simulated to complete the intervention of human skeletal muscle spasm.2.Successfully cultured normal human skeletal muscle cells and injured human skeletal muscle cells.3.Successfully produced three-dimensional human skeletal muscle cell gel.4.By detecting total superoxide dismutase,malondialdehyde and creatine kinase,there wassignificant difference between the C group and the injury group(P < 0.05),but no significant difference compared with the normal group(P > 0.05).The best pressure is 0.938 MPa.5 The p-LATS1,p-MST1,and p-Yap indicators were detected by western blot.It was concluded that the skeletal muscle cells of the injured human skeletal muscle cells were opened by the sputum method.The untreated human skeletal muscle cells will open the cells.The death pathway,the injury description method is related to the Hippo/Yap pathway.Conclusion: 1.The optimal pressure of rolling manipulation is 0.938 MPa.2.The injured human skeletal muscle cells opened the Hippo/Yap pathway after sputum intervention. |
