Basic Studies On Reducing Toxictity And Keeping Efficacy Of The Seeds Of Annona Squamosa And Biotransformation Of Annonaceous Acetogenins

The technology of fungal fermentation has been applied to Chinese medicine research,which can change the contents of main compounds in Chinese medicine,or the structure of effective constituents,widen the scope of medicine source,improve the efficacy of Chinese medicine,and may reduce its toxicity.This dissertation is a basic research on the seeds of Annona squamosa for keeping efficacy and reducing toxicity,which are divided into several part following for studying.Chapter 1 Literature reviewSystematically summarizes about the research of main chemical components,pharmacological activity of Annona squamosa were given.Besides,A detailed outline of medicinal fungi fermentation engineering containing solid and liquid fermentation technology and its application in traditional Chinese medicine research was mentioned in this part.Chapter 2 The effect of fungal fermentation on adaptability of the seeds of Annona squamosa and the contents of total annonaceous acetogeninsFive medicinal fungi was selected originally,which were based on previous research experience,and combined with the characteristics of strain growth itself.According to the activation time and growth of medicinal fungi,four kinds of medicinal fungi were selected for the fermentation of Annona squamosa,such as Ganoderma lucidum.The results showed that four medicinal fungi selected could grow better in the seeds matrix,and the contents of total annonaceous acetogenins improved.Besides,there were effects of different degree on the contents between different types of annonaceous acetogenins.Chapter 3 The content of total annonaceous acetogenins and acute toxicity of different extracts of the seeds of Annona squamosaThe content of total annonaceous acetogenins in 95%alcohol extract from the seeds of Annona squamosa was determined,while the contents in petroleum ether extract,chloroform extract,ethyl acetate extract and n-butanol extract were also determined.The results showed that there were large amounts of low polarity annonaceous acetogenins existing in petroleum ether extract.The acute toxicity tests of 95%ethanol extract and petroleum ether extract were taken,which found more toxicity in 95%ethanol extract.It might be concluded that the presence of large amount of low polarity annonaceous acetogenins were related to the decrease toxicity of petroleum ether extract.Chapter 4 Antioxidant activity about medicinal fungi fermentation on the seeds of Annona squamosa in vitroThe changes of antioxidant activity of Annona squamosa before and after fermentation were investigated in vitro by using DPPH free radical scavenging rate and reducing Fe3+capacity models.The results showed that the antioxidant activities of AA,DA and FA mycoplasma were better,while VA mycoplasma took second place.It was concluded that the increased antioxidant activity might be related to the increase contents of total annonaceous acetogenins,or the new potential antioxidant activity constituents.Chapter 5 Anti-tumor activity and toxicity about medicinal fungi fermentation on the seeds of Annona squamosa in vitroNormal liver cells named LO2 and kidney cells named HK-2 were selected to investigate the toxicity of the seeds of Annona squamosa in vitro before and after fermentation.The results found that the toxicity on liver and kidney cells decreased after fermentation.Anti-tumor activity in vitro of the seeds of Annona squamosa before and after fermentation were evaluated by estimating the inhibition ratios of five tumor cell lines named HepG2,SMMC-7721,HeLa,MCF-7 and A549 cells.The results showed that each mycoplasma could still keep its original anti-tumor activity in vitro.Besides,each group had strdhg inhibition ratio of breast cancer cell named MCF-7.Chapter 6 Anti-tumor activity and acute toxicity about medicinal fungi fermentation on the seeds of Annona squamosa in vivoThe toxicity tests of Annona squamosa before and after fermentation(fermented by four different medicinal fungi designated as A,V,D,F)were carried out by taking acute tocity tests on mice.The results found that the toxicity of VA and AA mycoplasma decreased significantly,while the toxicity of DA and FA mycoplasma reduced slightly and unobviously.Antitumor activities of Annona squamosa before and after fermentation were assessed by taking the transplanted sarcoma S180 tumor-bearing mice model in vivo.The results found that VA and AA mycoplasma had better antitumor activity than before,while they maintained or improvd the immunity of mice.However,anntitumor activity of DA and FA mycoplasma decreased.Besides they could damage the immune organs of mice.It was concluded that the decrease antitumor activity of DA and FA mycoplasma might be related to their damages on the immune organ of mice.Therefore,V and A fungi might be excellent strains for the study of keeping efficacy and reducing toxicity of Annona squamosa.Chapter 7 Biotransformation of annonaceous acetogenin named annosquatin-I by V fungus liquid-state fermentationThe substrate,a distant double tetrahydrofuran ring type of annonaceous acetogenins named annosquatin-I,was used by V strain liquid-state fermentation.The results showed that V strain could make annosquatin-I transformate structure to generate new material,which contained N element.Besides,fracture might occur between two tetrahydrofuran ring of original substrate.It was supposed that the new material might be related to good effects and low tocity of VA mycoplasma.