Effects Of Angelicanaphtha On CaN Signal Pat-hways And T-type Calcium Channel Of Angio-tensinⅡ-induced Hypertrophy Cardiomyocy-te

Objective:To explore the mechanism of Angelicanaphtha (AN) on AngiotensinII(AngII)-induced cardiac myocyte hypertrophy. By observing impact on calciumconcentration, cell Cycle, mitochondrial membrane potential changes and the proteinexpression of CaN, Cav3.1, Cav3.2, Caspase-12and Caspase-3induced by AngII inthe myocardial cell.Methods:Cells with concentration of1x106/mL were cultured in closed incubator with37℃and5%CO2for72h. Cardiomyocyte hypertrophy model was established by10-4mg·L-1AngII treatment. AN was made by CO2supercritical fluid extraction.Subjects were divided into5groups: normal control group, model group (myocardialcell with AngII, final concentration10-4mg·L-1), AN low-dose intervention group(treatment myocardial cells with AN of5mg·L-1for24h); AN middle-doseintervention group (treatment myocardial cells with AN of10mg·L-1for24h); ANhigh-dose intervention group(pretreatment myocardial cells with AN of20mg·L-1for24h); the cell viability of cardiomyocytes was detected by MTT assay; themitochondrial membrane potential and calcium concentration were detected with aconfocal microscopy; the cell cycle was detected by flow cytometry; the proteinexpression of CaN, Cav3.1, Cav3.2, caspase-12and caspase-3were detected bywestern blotting assay.Results:1. MTT assay for myocardial cell viability detection: Compared with the normalcontrol group, cell viability of myocardial cells was significantly increased in modelgroup (P＜0.05). Compared with the model group, the cell viability was significantlydecreased in all AN groups (P＜0.05).2. Calcium concentration of myocardial cells: Compared with the normal controlgroup, the calcium concentration of myocardial cells was significantly increased in model group (P＜0.05). Compared with the model group, the calcium concentrationof myocardial cells was significantly decreased in all AN groups (P＜0.05).3. Confocal laser scanning results: Compared with normal control group theRh123fluorescence intensity of myocardial cells in the model group was very high,but in AN interventionn group was significantly reduced.(P＜0.05).4. Flow cytometry results: Compared with normal control group the ration of G2phase cells in the model group was significantly increased, but in AN interventionngroup was significantly reduced.(P＜0.05).5. Western blot results: There was a certain amount of expression of CaN, Cav3.1,Cav3.2, caspase-12and caspase-3protein in normal control group. These proteinsexpression in model group significantly was increased(P＜0.05). In AN interventiongroup, CaN, Cav3.1, Cav3.2protein expression was significantly decreased(P＜0.05),caspase-12and caspase-3protein expression was significantly lower(P＜0.05).Conclusions:1. Angelicanaphtha can treat myocardial cell hypertrophy by blocking ofCa2+influx and inhibiting myocardial apoptosis.2. This study confirmed the angelica naphtha from the micro for blood gasmedicine and traditional Chinese medicine theory of qi and blood.