Anti-hepatoma Efficacy And Pharmacokinetics Research On Active Component Of Chiisanoside Isolated From Acanthopanax Sessiliflorus

Acanthopanax sessiliflorus is a Chinese edible herbal plant which is rich in northeast medicinal plant resources,and is commonly grown in some areas such as Jilin,Liaoning and Heilongjiang Province,it also can be found in Japan and South Korea.Chiisanoside is one of the tritererpenoids separating from Acanthopanax sessiliflorus.Modern pharmacology study indicated that chiisanoside was widely used in anti-rotaviral,anti-inflammation,the inhibition of lipase and the therapeutic effect of diabetes.Optimization of extraction process for chiisanoside in Acanthopanax sessiliflorus leaves,activities of anti-tumor and pharmacokinetics were investigated in this study.Extraction process of chiisanoside from Acanthopanax sessiliflorus was optimized by response surface methodology.On the basis of single factor tests,the effects of ethanol concentration,extraction time and the ratio of material to solvent were studied to increase the extraction rate by Box-Behnken.The optimal extraction conditions were as follows: ethanol concentration 72%,extraction time 40 min,solid to liquid ratio 1:118.Under the optimum conditions,the extraction yield of chiisanoside was 3.56%.The effective component from the leaves of Acanthopanax sessiliflorus was purified by macroporous resin.Explore 7 macroporus resins by using the test of static adsorption and desorption,and optimize the process parameters.The D101 macroporous resin was found to have better adsorption and desorption.The optimum conditions for purification process were as follows: the sample concentration was 0.2 g/mL,sample volume was 5 times of column volume;50% ethanol with 7 times of column volume.It was the first time to combine the two methods above to optimize extraction process of chiisanoside.On the basis of that,5 kg leaves of Acanthopanax sessiliflorus were extracted to seprate chiisanoside.9.58 g chiisanoside was isolated by using column chromatographic.As for anti-tumor activities,the results revealed that chiisanoside had significant inhibition on H22 ascitic hepatic tumor cell growth with no side effects on immune organs.The serum cytokines of IL-2,TNF-α,IFN-γ were all enhanced after treatment,whereas VEGF was inhibited in serum.Moreover,from the observations of H & E,TUNEL staining and immunohistochemical analysis,it can be found that chiisanoside had the positive effects on inducing apoptosis.The results showed that chiisanoside treatment displayed the up-regualation of Bax and caspase-3,and the down-regualation of the Bal-2 and VEGF.Futher investigations on pharmacokinetics and tissue distribution revealed that chiisanoside had a trait of fast absorbtion.Furthermore,tissue distribution showed that the highest level was observed in liver and small intestine,which indicated that chiisanoside showed good liver and small intestine-targeting efficiency.