| Obejective:To provide new cell biophysics experimental and theoretical basis For acupuncture and massage’s basic mechanical action,through the study of the effects of integrin-cytoskeleton signal transduction pathways in vitro of different tension stress friction stimulation on Fascial connective tissue fibroblast related meridiansMethod:Enzyme digestion method is used to obtain the Fascial connective tissue fibroblast related meridians in vitro, Develop from Fascial connective tissue fibroblast related meridians cell biology experiment platform after identification, and stimulate it with tension stress and friction, and observe the distribution of fibroblast integrin β1using fluorescence microscope and qualitative semi-quantitative analysis, and observe microfilament morphological changes of fibroblast.Results:(1) The research on the effects of integrin β1in vitro of different tension stress friction stimulation on Fascial connective tissue fibroblast related meridians all have significant difference. ①Stress stimulation:A single stimulation in each group, the fluorescence intensity of50kpa group is not obvious change compared with blank control group;the fluorescence intensity of100kpa and200kpa group is better than blank group;Multiple stimulation in each group,50kpa group of the fluorescence intensity is the brightest,100kpa group is better, the fluorescence intensity of200kpa group is better than blank group too。②Friction stimulation:A single stimulation in each group,0.5Hz group of fluorescence intensity not obvious change compared with blank control group, the fluorescence intensity of1Hz and2Hz group is better than blank group;1Hz group of fluorescence intensity is the brightest;Multiple stimulation in each group, lHz group of fluorescence intensity is the brightest, the fluorescence intensity of1Hz group is not obvious change compared with blank control group, the fluorescence intensity of2Hz group is darker than blank group.③Tension stimulation:A single stimulation in each group, with the increase of stimulus intensity, the fluorescent intensity gradually brighter, the fluorescence intensity of3000μ strain group is the brightest;Multiple stimulation in each group, the fluorescence intensity of2000μstrain group is the brightest,1000μ strain group is better, the fluorescence intensity of3000μ strain group is not obvious change compared with blank control group.(2) The research on the effects of microfilament morphological changes in vitro of different tension stress friction stimulation on Fascial connective tissue fibroblast related meridians all have significant difference. ①Stress stimulation:A single stimulation in each group,50kpa and100kpa group visible microfilament and nuclei are deepened, microfilament lined with blank group.no significant difference between visible microfilament and nuclei are200kpa group was obviously deepened, microfilament lined with blank control group still has no obvious difference;Multiple stimulation in each group,50kpa group of microfilament and nuclei are deepened, microfilament distribution is relatively scattered, rare throughout the cell length,100kpa microfilament group compared with blank control group and the nuclei are hyperchromatic obviously, microfilament fusion obvious solutions for short,arrangement without rules, slightly dotted in intracellular for nearly myosin after depolymerization, cell membrane may be seen occasionally filopodia200kpa multiple nuclei and microfilament punctuate dye strength group, significantly depolymerized microfilament, arrangement without rules, there are so many dotted in intracellular for nearly myosin after depolymerization, peripheral cell membrane highly irregular, numerous filopodia.②Friction stimulation:A single stimulation in each group,0.5Hz and1Hz single friction stimulation group compared with blank control group changed little,visible microfilament and nuclei are deepened,microfilament lined with blank group, no significant difference between group2Hz visible microfilament and nuclei are obviously deepened, microfilament lined with blank control group scattered, Multiple stimulation in each group,0.5Hz microfilament group compared with blank control group and the nuclei are deepening microfilament distribution is relatively scattered, microfilament fusion short, dotted in intracellular, did not see filopodia, the1Hz group significantly hyperchromatic nuclei and microfilament, microfilament fusion obvious solutions for short, arrangement without rules, appears dotted in intracellular for nearly myosin after depolymerization, cell membrane may be seen occasionally filopodia, group2Hz nuclei and microfilament punctuate dye, microfilament depolymerization, arrangement without rules, there are so many dotted in intracellular.③Tension stimulation:A single stimulation in each group,1000μ strain group is the nucleus and microfilament dyeing, slightly disorderly arranged microfilament compared with blank control group;2000μ strain group is the nucleus and microfilament dyeing,microfilament arranged disorderly, fusion obvious solutions for short,arrangement without rules;3000μ strain group is the nucleus and microfilament dyeing, microfilament arranged disorderly, microfilament fusion obvious solutions for short, arrangement without rules, point-like particle duality can be distributed in intracellular; Multiple stimulation in each group, the1000μ strain group is the nucleus and microfilament dyeing, microfilament fusion obvious solutions for short, arrangement without rules, slightly dotted in intracellular for depolymerization nearly myosin after;2000μ strain group is the nucleus and microfilament dyeing, microfilament fusion obvious solutions for short, arrangement without rules, slightly dotted in intracellular, appear along the direction of mechanics of cell microfilament away from the nucleus;3000μstrain group is the nucleus and microfilament dyeing, microfilament fusion obvious solutions for short, arrangement without rules, slightly dotted in intracellular, appear along the direction of mechanics of cell microfilament away from the nucleus.Conclusion:The expression of integrin and microfilament morphological changes in vitro of different tension stress friction stimulation on Fascial connective tissue fibroblast related meridians.(1)Common regularity and discriminative features of the expression of integrin in vitro of different tension stress friction stimulation on Fascial connective tissue fibroblast related meridians.①Common regularity:A single stimulation in each group, the integrin β1express higher under moderately or severe stimulus; Multiple stimulation in each group, the integrin β1express higher under mild or moderate stimulus.②Discriminative features:No matter single or multiple stimuli, integrin expression under external force stimulation quantity increased overall is a trend, but multiple severe friction stimulation, integrin expression is relatively blank flip, integrin expression is restrained, decreased expression.(2)Common regularity and discriminative features of the microfilament morphological changes in vitro of different tension stress friction stimulation on Fascial connective tissue fibroblast related meridians.①Common regularity:No matter single or multiple stimuli, microfilament morphology change from gradient features, and the gradient features’changes of multiple stimulation is more obvious than single stimulation. ②Discriminative features:Microfilament morphology change under tension stimulation is more obvious than stress and friction.(3) the regularity of integrin expression microfilament morphology change may reveal the basic physics signal and transmission principle of cell biology of acupuncture and massage’s basic mechanical action on Fascial connective tissue fibroblast related meridians from the perspective of cell mechanics. The clinical significance of the acupuncture massage may be:①From the perspective of signal feelings:in the clinical treatment, treatment for a short period of time, may be suitable to use heavy technique, signal feeling more intense; Long time of treatment may be appropriate use of lighter, keep the feeling of the acupuncture point at a higher level.②From the perspective of signal transmission:The signal transmission efficiency is higher that can make the meridians by tension technique. |
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