Efficacy-enhancing And Toxicity-reducing Mechanism Of Compatibility Of Ginsenoside Rb₁ And Aconitine

Objective:To study the efficacy-enhancing and toxicity-reducing mechanism of compatibility of ginsenoside Rb₁ and aconitine.Methods:1. Use propafenone hydrochloride to establish the acute heart failure rats model by bolus injection. Positive drug Sheng Mai Injection, ginsenoside Rb₁, aconitine, compatibility of ginsenoside Rb₁ and aconitine were intravenous injected into the rats, then the hemodynamic index was recorded by using BL-420 system, in addition, the concentration of Ang-I, ANP, ET, ALD, TNF-a in blood serum were measured by using ELISA assay; 2. The rat myocardial cells were obtained by trypsin digestion and purified by different velocity of adherence, pentobarbital sodium was used to establish the heart-failure cell model, the drug treatment time was 1 hour, lastly, the vitality of the myocardial cells, ATP enzyme and the concentration of relative ions were measured.3. The myocardial cells were treated with ginsenoside Rb₁, aconitine ?0.2%?, compatibility ginsenoside Rb₁ and aconitine for 1 hour, afterwards, the vitality of the myocardial cells, ATP enzyme and the concentration of relative ions and the content of glycogen, succinate dehydrogenase, cycochrome C oxidase, lactate dehydrogenase were measured.4. Total RNA was extracted from myocardial cells after drug treatment, then RT-PCR was performed to detect mRNA level of Kv4.2, Nav1.5, in addition, total protein was extracted to detect the expression of Kv4.2 and Nav1.5 protein using western blot.Results:1. The impact on rats of acute heart failure:After 5 min treatment, compared with the model group, compatibility of ginsenoside Rb₁ and aconitine ?1:1, 2:1? groups increased frequency particularly in rats, each group could significantly increase the value of+LV dp/dtmax, the group of ginsenoside Rb₁ increased the value of-LV dp/dtmax significantly; the group of Shenmai injection and ginsenoside Rb₁ and compatibility could reduce the content of Ang-I, especially the group of compatibility of ginsenoside Rb₁ and aconitine?2:1?; the group of ginsenoside Rb₁, aconitine, compatibility of ginsenoside Rb₁ and aconitine ?1:1? reduced the content of ANP, compatibility of ginsenoside Rb₁ and aconitine ?1:1,2:1,1:2? groups reduced the content of TNF-a, compatibility of ginsenoside Rb₁ and aconitine ?2:1,1:2? groups increased the content of ALD, the group of ginsenoside Rb₁, compatibility of ginsenoside Rb₁ and aconitine ?1:2? reduced the content of ET, especially the group of ginsenoside Rb₁ showed the most potent effect 2. The impact on heart-failure cells: aconitine (1×10^-7 mol·L^-1) and ginsenoside Rb₁ (1×10^-7 mol·L^-1) and its compatibility increased the viability of heart-failure cells caused by pentobarbital sodium, compatibility of ginsenoside Rb₁ and aconitine ?1:2? group could significantly reduce the content of ACP, LDH. Aconitine (1×10^-7 mol·L^-1) and ginsenoside Rb₁ (1×10^-7 mol·L^-1) and its compatibility groups could increse the concentration of Na⁺and Mg²⁺, reduce the concentration of K⁺and Ca²⁺, especially compatibility of ginsenoside Rb₁ and aconitine ?1:2,2:1?group showed the most potent effect;increase the myocardial intracellular Ca²⁺-ATP enzyme activity and Ca²⁺-Mg²⁺-ATP enzyme activity, reduce Na⁺-K⁺-ATP enzyme activity, especially compatibility of ginsenoside Rb₁ and aconitine ?1:1,2:1?group showed the most potent effect.3. The impact on the normal cardiomyocytes:Aconitine?0.2%? could injure cardiomyocytes, compatibility of ginsenoside Rb₁(1×10^-6 mol·L^-1)and aconitine?0.2%? could significantly enhance myocardial viability, weaken effect of ion concentration caused by aconitine?0.2%?, particularly, the concentration of Na⁺ reduced significantly; on the other hand, myocardial intracellular Ca²⁺-ATP enzyme activity, Na⁺-K⁺-ATP enzyme and Ca²⁺-Mg²⁺-ATP enzyme activity, the effect of compatibility of ginsenoside Rb₁ and aconitine ?1:2,2:1?groups were remarkably improved; compatibility of ginsenoside Rb₁ and aconitine ?1:1,1:2,2:1?groups could also slightly rescue the abnormal content exchange of glycogen, succinate dehydrogenase, cytochrome C oxidase and lactate dehydrogenase caused by aconitine?0.2%?, compatibility of ginsenoside Rb₁ and aconitine ?2:1?group obviously increased the content of glycogen, reduced the content of lactate dehydrogenase; compatibility of ginsenoside Rb₁ and aconitine ?1:2?group obviously reduced the content of cytochrome C oxidase.4. The impact on expression of Kv4.2, Nav1.5:Cedilanid group, compatibility of ginsenoside Rb₁ and aconitine ?1:2,2:1?groups enhanced the expression level of Nav1.5mRNA in heart-failure cells; Cedilanid group, compatibility of ginsenoside Rb₁ and aconitine ?2:1?group enhanced the expression level of Kv4.2mRNA in heart-failure cells. Aconitine group and compatibility of ginsenoside Rb₁ and aconitine ?1:1,2:1?groups tended to inhibit the protein expression of Nav1.5 in heart-failure cells; each group had no significant differences on Kv4.2 protein expression in heart-failure cells. Aconitine ?0.2%? inhibited the expression level of Kv4.2mRNA in myocardial cells, ginsenoside Rb₁ group and compatibility of ginsenoside Rb₁ and aconitine ?1:1,1:2, 2:1?groups enhanced the expression level of Kv4.2mRNA in myocardial cells; each group had no significant differences on Nav1.5mRNA expression level. Aconitine ?0.2%? enhanced the protein expression of Kv4.2 and Nav1.5 in myocardial cells, compatibility of ginsenoside Rb₁ and aconitine ?1:1,1:2,2:1?groups tended to reduce the protein expression of Kv4.2 and Nav1.5 in myocardial cells.Conclusions:1. Compatibility of ginsenoside Rb₁ and aconitine in high concentration?0.2%? could reduce cardiac toxicity caused by aconitine in high concentration. Its mechanism was related to improving the stability of cells membrane, improving celluar ion home -ostasis by regulating the expression of ion channel proteins and activity of ion exchange ATP enzyme, improving cellular energy metabolism.2. Compatibility of ginsenoside Rb₁ and aconitine in high concentration(1×10^-7 mol·L^-1) could enhance treatment effect on heart failure. Its mechanism was related to improving cardiac function, regulating the nervous cytokine secretion of system RAAS.3. Efficacy-enhancing and toxicity-reducing mechanism of compatibility of ginsenoside Rb₁ and aconitine was associated with the nervous cytokine secretion, structure change and function change in cells. Its effect was also associated with the drug concentration used and the specific body state.