The Effect And Preliminary Mechanism Study On Monkshood Polysaccharide Combined With Aconitine To The Hepatocellualar Carcinoma Cell

The incidence of malignant tumors is increasing in recent years all over the world, among which the mortality of HCC is ranked third. In traditional Chinese medicine, strengthening body resistance and eliminating evil is the basic law of antitumor, and aconite is applicable in reviving yang for resuscitation, tonifying fire and helping yang and eliminating cold to stop pain. Recent research has shown that aconite decoction has the effect of anticancer, strengthening heart, antiaging and enhancing immunity, the main effective components of which are monkshood polysaccharide and aconitine. In the present study, the effect and preliminary mechanism of Monkshood Polysaccharide combined with aconitine to the hepatocellualar carcinoma cell was investigated in order to provide evidence for rational clinical application of aconite.Objective:To study the immune effect and immunity anti-hepatoma (HCC) effect of Monkshood Polysaccharide; the anti-hepatoma effect and the damage to the immunity of aconitine; the anti-hepatoma effect of Monkshood Polysaccharide combined with aconitine and their ratios alter, so as to rate the rationality of the Pro bed aconite "reuse", "reuse+long fried".Methods:The fluorescence staining method was used to observe the apoptotic effect of hepal-6 hepatoma cells induced by Monkshood Polysaccharide, aconitine and their combination with different proportions; MTT method was used to observe the different effects of hepal-6 cells, the proliferation of spleen cells and spleen cells killing hepatoma carcinoma hepal-6 cells by Monkshood Polysaccharide, aconitine and their combination with different proportions; Model of C57 tumor-bearing mice was established to observe the tumor inhibitory rate and the effect of immunity by Monkshood Polysaccharide, aconitine and their combination with different proportions.Results:1. Monkshood Polysaccharide has no significant effect on the proliferation and apoptosis of Hepal-6 hepatocellular carcinoma cells (P>0.05),it can promote the proliferation of mouse spleen cells in vitro (P<0.01), and has a dose-dependent relationship,it can also enhance the killing ability of spleen lymphocytes to the hepatocellular carcinoma cells;Monkshood Polysaccharide has a weak inhibitory effect on Tumor(tumor weight) of the mice bearing Hepal-6 (inhibition rate 18.22%),can increase the spleen weight and spleen index of C57 mice bearing the tumor (P<0.05); significantly increase the quantity of CD4+and CD8+in spleen T cells (P<0.05).2.Aconitine has an obvious inhibitory effect on the proliferation of Hepal-6 cells (P<0.05), in both dose-dependent and time-dependent manner, it can inhibit the proliferation of mouse spleen cells cultured in vitro(P<0.01),in. a certain dose-dependent manner,it can promote the apoptosis of Hepal-6 cells, and has a dose-dependent relationship; The aconitine shows some inhibitory effect on the tumor (tumor weight) of mice bearing Hepal-6 tumor, in dose dependent manner,it has a certain trend of the inhibitory effect on the spleen weight,spleen index, thymus weight and thymus index of mice bearing tumor, but there is no significant effect (P>0.05).3. The Invalid concentration of the Aconitine (50μg·mL-1) had no inhibitory effect on the proliferation of Hepal-6 (hepatocellular carcinoma cells), according to the combining use in 1:10 proportion of Monkshood Polysaccharide is not also obvious inhibitory effect, the effective concentration of aconitine (200 and400μg·mL-1),combined with Monkshood Polysaccharide in the proportion of 1:10, doesn’t increase the inhibitory effect on the Hepal-6; the effective concentration of aconitine (200μg·mL-1),combined with Monkshood Polysaccharide in the proportion of 1:10, doesn’t increase the inhibitory effect on the Hepal-6, when the proportion of Aconitine to Monkshood Polysaccharide is evident 1:20, the inhibition rate is increased; aconitine in effective concentration(200μg·mL-1) is combined with Monkshood Polysaccharide in the proportion of 1:5、1:10、1:20, along with the increasing mixture of Monkshood Polysaccharide, the inhibitory effect of Aconitine diminished; it can also enhance the killing ability of spleen lymphocytes to the hepatocellular carcinoma cells;aconitine in the effective dose(150μg·kg-1) is combined with Monkshood Polysaccharide in the proportions of 1:10, 1:20,it shows inhibitory effect on the tumor (tumor weight) of mice bearing Hepal-6 tumor, but no further effect when in the proportions of 1:40; aconitine in the effective dose(150μg·kg-1) s combined with Monkshood Polysaccharide in 3 different proportions (1:10,1:20 1:40 Aconitine:Polysaccharide), the spleen, spleen index,the thymus and thymus index of the mice bearing Hepal-6 tumor can be promoted, it can increase the number of macrophages in the spleen,and the number of CD4+ and CD8+ in the spleen T cells is significantly increased.Conclusion:Monkshood Polysaccharide is capable of enhancement of immune fuction, which has played a role in antitumor immunity within the range of the effective doses, and has no direct inhibition to hepatocellular carcinoma (HCC). While aconitine has the capacity of direct antitumor effect and immune injury in tumor-bearing mice. The combination of Monkshood Polysaccharide and aconitine can strengthen the anti-tumor effect, improve the immune injury induced by aconitine in Hepal-6 tumor-bearing mice, and the change proportion of which has certain influence on anti-tumor effect.