| Objective:To observe the effects that the compatibility liquiritin?glycyrrhetinic acid and hypaconitine to the damaged myocardial cells and to discuss the attenuation and synergy effect mechanism.Methods:Using primary cultured myocardial cells as research subjects,through bioche-mical and genetic level research methods to explain the mechanism from different levels and perspectives.Using MTT method to study the effect that diifferent concentration and different time po-ints of hypaconitine to the heart failure model,and find out the concentration and the time po-int that aconitine works best(T0 C0).Then according the result set up the proportion of low?m iddle?high liquiritin?glycyrrhetinic acid to hypaconitine respectively groups.Cluture the pr-imary myocardial cells in vitro and observe the morphological changes of the myocardial cel-ls by using inverted microscope,from the side of morphology to observe the changes of norm-al myocardial cells?myocardial of C0 hypaconitine and diffrent compatibility groups.MTT assay was used to detect the hypaconitine,Liquiritin,glycyrrhizic acid effects on the activity of myocardial cells and the survival rate.Learn the changes of MDA vigour of SOD?GSH-PX under different concentration compatibility liquiritin?glyc-yrrhetinic acid and hypaconitine.According the result to select significative compatib-ility groups,using RT-PCR methodolo-gy to delect the level of intracellular connexin 43(Cx43)gene transcription.Result:(1)Morphological observation:After 2h hypaconitine effects myocardial cells show ed a contraction of pseudopodia,vacuoles,morphological changes of particles.480?mol/L Liquir-itin,120?mol/L glycyrrhizic acid group less contraction of pseudo-opodia,cytoplasmic vacu-of ization and the appearance of particulate matter was not obvious.myocardial cells grow w-ell.(2)The effects on myocardial cell viability.myocardial cell injuryo and xidation injury:The time and concentration of hypaconitine effect myocardial cells work best were 2H and 120?mol/L by MTT.Hypaconitine could cause myocardial cells viability decreased and injure cells toxicixy,myo-cardial viability was about 71%(P<0.01);Hypaconitine groups MDA levels were signific-antly higher than others,significant increase in membrane permeability(P<0.01);Hypaconi-tine groups MDA levels were significantly lower than others(P<0.01);Low,med ium and high doses of Liquiritin,glycyrrhizic acid after hypaconitine compatible res-pectively groups could improve myocardial viability,increased membrane permeability,improved oxidation injury and reduce the lever of apoptosis(P<0.05),Which the high dose of Liquiritin,the low dose of glycyrrhizic acid compatibility hypaconitine group most obvious effected(P<0.01).(3)The effects on intracellular connexin 43(Cx43)gene transcription:Compared with the control group,hypaconitine group Cx43 levels were significantly increased gene transcription(P<0.05),liquiritin,glycyrrhizic acid after hypaconitine compatible respectively groups could reduce the hypaconitine-induced Cx43 gene transcription increase,And liquiritin compatibility hypaconitine groups more significant effected.Conclusions:Liquiritin,glycyrrhizic acid groups had a significant effects in various aspects,which the antagonism of hypaconitine induced myocardial cell viability decreased and to reduce the MDA content increase by hypaconitine induced myocardial injury and improved vigour of SOD?GSH-PX.Experiments showed that high dose of liquiritin,low dose of glycyrrhizic acid after hypaconitine combination respectively had the best effects in reducing myocardial injury induced by hypaco-nitine,Its possible way to achieve was to improve the hypaconitine-induced cardiac muscle cell membrane permeability changes and influences of Cx43 gene transcription. |
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