ICC Model Ca²⁺ Regulation Mechanism Of Dampness Block Middle Energizer And Pingwei San Interfere Effect Experimental Researching

Objective:Establishing the dampness block middle energizer model of ICC, searching the Ca2+ regulating mechanism of ICC dampness block middle energizer model as researching target spot. Revealing the mode of effect by Pingwei San treats damp blockage middle energizer to regulate gastrointestinal motility.Methods:â‘ Establishing the damp blockage middle energizer model of rats: simulating the pathogen of dampness block "Living dampness, improper diet, abnormal hunger and full, sleep reduce, irregular emotion", establishing the rat model which created by Huangxiu Shen.â‘¡Preparing the model blood serum after 20 days molding, grouping randomly the rest model rats, dose group feed Pingwei San, natural and blank group feed 0.9%normal saline for 3 days. Preparing each group blood serum. Culturing in vitro ICC of rat's small intestine. Confirm the cells by morphology and immunofluorescence technic, then built the ICC damp blockage middle energizer model by damp blockage middle energizer model rats'blood serum.â‘¢Apply serum pharmacology observe group difference of serum and Ca2+ contact closely to ICC regulation mechanism factor.Result:â‘ ICC orphology observing and immunofluorescence technic confirm: Under the inverted microscope, culturing in vitro original generation ICC's special morphological characteristics.24h Cells were adherence and deformed, presenting triangle or fusiformis, a few extend out. The 3rd day formed long neurite like axons.7 days later, ICC can extend out second, third grade neurite. The neurite can connect into complicated net structure which is the basic of slow wave function (Figure). Under the fluorescence microscope, the surface c-kit of ICC presents positive, soma and neurite are colored significant, a few extend out long neurite.â‘¡ICC model establishment:model cells'shape change obviously which become flat, synapse connect closely, cell gap smaller, net connection not obvious. Objective indicator shows compared with normal ICC, Ca2+ concentration inner and outer model cell descend (p<0.05), Ca2+-ATP descend(p<0.05),PLC descend (p<0.05), IP3R descend (p<0.05), model ICC and normal ICC have significant difference, model ICC establish successfully.â‘¢Comparing with normal group, model group Ca2+ concentration inner and outer cells both descend(p<0.05), PLC rise(p<0.05), IP3 has no significant difference, IP3R descend (p<0.05); Comparing with normal group, natural cure group Ca2+ concentration inner and outer cells has no significant difference, Ca2+-ATP rise (p<0.05),PLC rise (p<0.05), IP3 has no significant difference,IP3R rise (p<0.05). Comparing with the model group, natural cure group Ca2+ concentration inner and outer natural cure group both rise (p<0.05), Ca2+-ATP rise (p<0.05), PLC rise (p<0.05), IP3 has no significant difference, IP3R rise; Comparing with the model group, Pingwei San Ca2+ concentration inner and outer cells both rise (p<0.05), Ca2+-ATP rise (p<0.05), PLC rise (p<0.05), IP3 rise (p<0.05), IP3R rise (p<0.05).Conclusion:â‘ Model ICC and normal ICC have significant difference, model ICC establish successfully.â‘¡Regulating Ca2+ concentration inner and outer cells through regulating cell Ca2+-ATP enzyme activity, making Ca2+ enter cells can expel avoiding Ca2+ in cells overload, on other hand through G protein coupling receptor-PLC-IP3 regulating Ca2+ concentration in cells, making Ca2+ concentration rise and transmit signal out of cells.â‘¢Inferring ICC is one of target spots which Pingwei San cure dampness block middle energizer DGIMD, Pingwei San cure dampness block middle energizer regulate Ca2+ in ICC mechanism and come the effect.