| Objective:To study the effect-enhancing and toxicity-reducing mechanism of ginsenoside Rg1 compatibility with aconitine in different concentrations on heart failure.Methods:1.Primary myocardial cells of natal rats were obtained by trypsin digestion and identification was performed by?-actinin immunofluorescence.2.Ginsenoside Rg1 compatibility of different concentrations of aconitine on normal cardiomyocytes.MTT method is used to detect the toxicity concentrations and the effective concentrations of aconoitine on the cardiomyocytes and the concentration of Ginsenoside Rg1.Then under the condition of different compatibility using the transmission electron microscope to observe the influence of the drug on the ultrastructure of cardiomyocytes;using the reagent kits to detect the cell membrane permeability and concentration of intracellular Na+,K+,Ca2+,Mg2+;testing the expression quantity of apoptosis related protein Bax,Caspase-3,and ion channel related protein Kv4.2,Nav1.5,RyR2 by using western blot;Real-Time PCR method for determining the relative expression of Kv4.2,Nav1.5 and RyR2 mRNA.3.The effects of Ginsenoside Rg1 compatibility of different concentrations of aconitine on the heart failure myocardial cells which is caused by Pentobarbital sodium.MTT method is used to detect the toxicity concentrations and the effective concentrations of Aconoitine on the myocardial cell and then observe the intracellular environment,ultrastructure and related protein expression as before.Results:1.The impact of ginsenoside Rg1 compatibility of different concentrations of aconitine on normal cardiomyocytes.Identified the cardiomyocytes are conform to the requirements of the experimental and the results showed that:compared with the control group,aconitine(3×10-3 mol/L)and aconitine(1.5×10-3 mol/L)can lead to a significant decrease viability of myocardial cells,damage of cellular membrane and organelles such as the mitochondrion,the endoplasmic reticulum,the nucleus and other ultrastructural damage.The two concentrations lead to increase of leakage ACP,LDH;increase the concentrations of Na+?Ca2+and decrease the concentrations of K+?Mg2+;increase the expression of Bax?Caspase-3?Kv4.2?Nav1.5 and RyR2 protein.At the same time,aconitine(3×10-3mol/L)elevated the expression of RyR2?Nav1.5 and Kv4.2 mRNA shows a significant cardiomyocytes toxicity.Compared with the corresponding concentration of aconitine group,ginsenoside Rg1 can reduce the above two toxic concentrations,reduce cell ultrastructural damage,reduce the leakage of ACP?LDH,decrease the concentrations of Na+?Ca2+and increase the concentration of K+,furthermore increase the expression of Nav1.5?Kv4.2 mRNA.In addition,aconitine(3×10-3mol/L)compatibility group,the expression of RyR2 mRNA decrease,aconitine(1.5×10-3mol/L)compatibility group decrease the expression of Bax protein.The rest of the concentrations have no effect on the vitality of the cells,ultrastructure,the vality of ACP and LDH,and ion concentrations.But aconitine(1×10-7mol/L?1×10-9mol/L)can just reduce the expression of Kv4.2 and Nav1.5 protein,compatibility group has increased after the trend,aconitine(1×10-7mol/L)increase the expression of Nav1.5?Kv4.2mRNA,after compatibility increased.2.The effects of ginsenoside Rg1 compatibility of different concentrations of aconitine on the Pentobarbital sodium induced heart failure cardiomyocytes.The results showed that:there is obvious toxicity on the model group cells when the concentrations of aconitine are 3×10-3mol/L and 1.5×10-3mol/L,cell viability decrease significantly,cell membrane,nucleus and cytoplasm of organelles damage;supernatant ACP,LDH increase,the concentrations of intracellular Na+and Ca2+increase and the expression of Nav1.5 and RyR2 protein increase.Compered with the corresponding concentrations of aconitine alone,ginsenoside Rg1 compatibility group can fight against the above indicators abnormal.Aconitine 1×10-5mol/L and the concentrations below can increase the vality of the heart failure cardiomyocytes,elevate the concentration of Na+,decrease the concentrations of K+?Ca2+,reduce the expression of Nav1.5 and Caspase-3,Bax protein.Aconitine 1×10-7mol/L can also elevate the expression of Nav1.5?Kv4.2mRNA;Compared with aconitine 1×10-7mol/L work alone,compatibility group can increase the expression of Nav1.5?RyR2 mRNA.After compatibility,aconitine1×10-9mol/L can decrese the expression of Nav1.5 mRNA.Conclusions:1.Ginsenoside Rg1 can not only reduce the toxicity of high does aconitine but also enhance the effect of low does aconitine on heart failure myocardial cells,shows toxicity-reducing and effect-enhancing.2.The mechanism of ginsenoside Rg1compatibility of aconitine toxicity-reducing and effect-enhancing on heart failure cardiomyocytes may be related to improving cardiomyocytes membrane stability,maintaining the ionic homeostasis and adjust the expression of iontransport related proteins and anti-apoptosis;moreover it's also related to the concentrations of compatibility. |
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