| Objective:According to the umbilical vein endothelial cells’changes of intracellular substances and energy regulating mechanism in accepting thermal stimulation, this paper study on heat energy to biological energy conversion mechanis of collaterals moxibustion.As experimentally demonstrated in cellular level, moxibustion thermal stimulation can spontaneously form "activating yang qi" and "promoting the body resistance and eliminating pathogenic factors"Methods:The use of human umbilical vein endothelial cells cultured in vitro builds a collateral system research platform. The direct stimulating effect of moxibustion on collateral system is simulated by different intensities of thermal stimulating Intervention on cell. Dynamic changes in cell morphology were observed under an inverted microscope. Application of enzyme activity spectrometry detects AMPK activity of cells. Applications of QPCR and western-bolt technology, tests the thermal stimulation of cell HSP70and the HSP90mRNA and protein expression. These results are analyzed and discussed the use of cytology knowledge, related with the traditional theory of Chinese medicine moxibustion.Results:1. Using an inverted microscope observed:Cells by thermal stimuli appear cellular edema and cytoplasmic granules of amyloidosis of cloudy swelling phenomenon immediately. Cloudy swelling weakens at the end of stimulation after6-8hours. The degree of cloudy swelling in40"C group is more obvious than43℃group’s.24hours after total stimulus ended,40℃group show little morphological differences with37℃, cloudy swelling loss, cell membrane edge of the granular material is reduced. While the43℃group of cells within the cytoplasm of color is37℃group was deepened, cell volume decreased, cell gap increases obviously. 2.Compared with37"C normal control group, the activity of AMPK in umbilical vein endothelial cells of40℃thermal stimulation group was obviously enhanced, and the activity of43℃group is the worst.40℃group is significant differences between the other two groups (P<005).3. Of40℃heat stimulation group, the expression on mRNA and protein of HSP70and HSP90are increased from the normal37"C temperature group, while in overheating damage group43℃, heat shock protein mRNA and protein expression is not markedly changes.40℃group with other differences between the two groups was statistically significant(P<0.05),except HSP90in43℃group.Conclusion:1.In the appropriate temperature stimulation can make endothelial cells’ AMPK, responsible for the energy supply path, activity was significantly elevated, content of heat shock protein, responsible for the protection and repair of the proteins, increased significantly. These changes not only the thermal tolerance of cells, and further regulate the functional structure of the cells in living matter and energy supply, so that the cells recovered in good condition to protect the cells play a normal physiological function. AMPK activation and HSP protein expression increased although two different boot mechanisms. But as important parts of the homeostasis regulation system, mutual have synergistic effect.2. Veri Tied by experiment, the heat energy to biological energy conversion mechanism caused by thermal stimulation on cells in the material and energy change is an object ive reality. This transformation depends on the cell through the self feedback formed in a certain range of homeostatic regulation mechanism Temperature threshold is the key to the cell feedback, determines the homeostatic adjustment mechanism start or not. It prompts us in clinical moxibust ion should pay attention to the quantity and degree of certainty 3. Through the moxibustion, cell dimensions that can spontaneously form "activating yang-qi"and "promoting the body resistance and eliminating pathogenic factors"... |
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