The Effect Of HYP On H9C2 Cells And L-type Calcium Channelin Hypericum Attenuatum Choisy

ObjectiveObserve the effect of Hyp on L-type calcium channel in myocytes.To investigate the mechanism of treating cardiovascular disease in Hypericum attenuatum choisy.Method1.Determination:To determine the content of hyperin and other flavonoids in Hypericum attenuatum choisy by HPLC.2.To investigate the effects of different hyperin concentrations of CCK-8 on the growth of H9C2 myocytes.3.Using the Whole-cell patch clamp technique to examine the effect of different concentrations of hyperin(5 μM,15 μM,50 μM)on L-type calcium channel currents.Results1.Determination:The average contents of hyperin,rutin,quercetrin,quercetin,kaempferol in samples were 0.13,0.19,0.09,0.28,0.14 mg/g by HPLC,respectively.2.The inhibition rate of different concentrations of H9C2 myocytes was determined by CCK-8.The myocytes survival rate increased with the increase of hyperin concentration.Based on the concentration of CCK-8,select three concentrations of 5μM,15μM,50 μM hyperin in L-type calcium channel current of H9C2 myocytes byWhole-cell patch clamp.The peak value of current density is dependent on the concentration of hyperin.At the maximum peak,at the 20mV voltage,The current densities of hyperin at the three concentrations 5 μM,15 μM,50 pM:-8.78 ± 2.43,-7.71 ± 1.80,6.59± 1.20.The current density decreases with the influence of concentration.The maximum peak value was positively correlated with hyperin concentration and the current intensity was negatively correlated with hyperin concentration at different voltages.The overturn potential is between+50mV-+60mV.In the curve of activation and inactivation,the semi-stable voltage is increased,and there is a positive correlation between the voltage and the concentration.In the recovery curve,the recovery time can be prolonged and there is a positive correlation with the concentration.Conclusions1.The flavonoids are rich in Hypericum attenuatum choisy.The content of five kinds of flavonoids is determined by HPLC method,which provides the basis for the determination of the content of the quality standard of Hypericum attenuatum choisy.2.Based on the effect of Hyp on cell proliferation,it is can infer that Hyp has a certain protective effect on H9C2.3.Hyp inhibited L-type calcium channel in H9C2 myocytes.4.Hyp is one of the bases of antiarrhythmic effect of Hypericum attenuatum choisy.