The Effects Of Huazhuo Jiedu Huoxue Tongluo Prescription Against Calcium Overload Mechanisms On The Mice With CIRI

Objective: To observe the effects of Huazhuo Jiedu Huoxue TongluoPrescription on the contents of Ca²⁺-ATPase/Na⁺-K⁺-ATPase/Glutamate（Glu） in the brain tissue, the contents of Calcineurin phosphatase （CaN） in theSerum and pathomorphology of CA1region in model mice with CIRI（Cerebral ischemia and reperfusion injury）. To evaluate the therapeutic effectof Huazhuo Jiedu Huoxue Tongluo Prescription, discuss the mechanism of thismethod on CIRI and confirm the protective effect on neurons of the mice withCIRI.Methods:50healthy male Kunming mice of specific pathogen free （SPF）were randomly divided into5groups: model group, high dose of HuazhuoJiedu Huoxue Tongluo prescription traditional Chinese Medicine （TCM）group （called “high dose group” as follows）, low dose of Huazhuo JieduHuoxue Tongluo prescription traditional Chinese medicine （TCM） group（called “low dose group” as follows）, Nimodipine control group （called“Nimodipine group” as follows） and the Sham operation group. Each grouphad10mice. We duplicated the model of CIRI mice with the operation ofcerebral ischem reperfusion through separating and ligating both side carotidarteries except sham-operation group. All mice were returned to their cagesafter the operation, ate food and water freely after models were successfullyestablished, and the drugs were given by gastric perfusion the next day.Physiological saline was given to the mice in the sham operation group andmodel group. Nimodipine solution was given to the mice in the Nimodipinegroup. High and low dose of Huazhuo Jiedu Huoxue Tongluo prescriptionChinese medicine decoection were given to the mice in the high dose groupand low dose group respectively. The drugs were given once a day and for14days continuously. The mice were cut head for obtaining cerebral tissue, the left part of which was fixed and cut into slices, the tissue was stained by HE（Hematoxylin） dye. We observed constitutional morphologic changes ofcerebral tissue under light microscope. Then we made the homogenate fromcerebral cortex tissue, took the supernatant after centrifuging, measured thecontent of the Ca²⁺-ATPase/Na⁺-K⁺-ATPase and Glu in the brain tissue andexamined the change of the serum CaN.Results:1. The morphological changes of nerve cells in CAI region:The pathomorphology under Light microscope indicated that Pyramidalcells in the hippocampus of the sham-operation group were arranged closely,cell nucleuses were big and round, nucleoli were clear. The layers of thepyramidal cells in the hippocampus of the model group were reduced, thepyramidal cells were arranged sparse and missed obviously, the cell nucleusesdiminished and the cell structure was vague. The scope of the pathologicalchanges in hippocampus of mice brain took on the pathological changes ofischem. The pathological changes of each medical group were improvedcompared with the model group.2. The effect of Huazhuo Jiedu Huoxue Tongluo Prescription on the content ofthe Ca²⁺-ATPase/Na⁺-K⁺-ATPase in the brain tissue of the CIRI mice:The activity of the Ca²⁺-ATPase/Na⁺-K⁺-ATPase in the brain tissue ofthe model group was lessoned compared with that of the sham-operationgroup （P<0.01）, the activity of the Ca²⁺-ATPase/Na⁺-K⁺-ATPase in the braintissue of the medical group was strengthened compared with that of the modelgroup （P<0.01）. The activity of the Ca²⁺-ATPase/Na⁺-K⁺-ATPase in the braintissue of high-dose group was higher than that of the low-dose group and theNimodipine group （P<0.01）. There was no different when other medicalgroups were compared with each other （P>0.05）.3. The effect of Huazhuo Jiedu Huoxue Tongluo Prescription on the content ofthe Glutamate in the brain tissue of the CIRI mice:The content of the Glutamate （Glu） in the brain tissue of the modelgroup was increased compared with that of the sham-operation group （P<0.01）, the content of the Glutamate （Glu） in the brain tissue of the medical group wasdecreased compared with that of the model group （P<0.01）. The content of theGlutamate （Glu） in the brain tissue of high-dose group was decreased than thatof the low-dose group and the Nimodipine group （P<0.01）. There was nodifferent when other medical groups were compared with each other （P>0.05）.4. The effect of Huazhuo Jiedu Huoxue Tongluo Prescription on the content ofthe CaN in serum of the CIRI mice:The content of the CaN in the serum of the model group was increasedcompared with that of the sham-operation group （P<0.01）, the content of theCaN in the serum of the medical group was decreased compared with that ofthe model group （P<0.01）. The content of the CaN in the brain tissue ofhigh-dose group was decreased than that of the low-dose group and theNimodipine group （P<0.01）. There was no different when other medicalgroups were compared with each other （P>0.05）.Conclusions:1The mice ligationed of bilateral carotid artery were the perfect models,because the pathomorphology under light microscope indicated that Pyramidalcells in the hippocampus were damaged.2The damage of the pyramidal cells in the hippocampus tissue is theimportant pathology change from CIRI. Huazhuo Jiedu Huoxue TongluoPrescription can alleviate the pathology damage caused by the cerebral ischemand have treatment function to the CIRI mice.3Huazhuo Jiedu Huoxue Tongluo Prescription can raise the activity of theCa²⁺-ATPase/Na⁺-K⁺-ATPase, lower the content of the Glu to Inhibition ofintracellular calcium overload resist the damage of the Excitatory amino acid.4Huazhuo Jiedu Huoxue Tongluo Prescription can lower the content ofthe CaN to inhibiting calcium channel signal caused apoptosis, and allevaitethe damage of the CIRI.5The improvement of neuropathology and biochemical markers in micewith CIRI proves that Huazhuo Jiedu Huoxue Tongluo Prescription has someproteetive effect on CIRI.