Studies On The Biological Active Constituents Of Heipiaoxiao

Tuanpiaoxiao,Changpiaoxiao and Heipiaoxiao are the eggs of Tenodera sinensis Saussure,Statilia maculata Thunberg,Hierodula patellifera Serville,and are collectively known as Ootheca mantidis,it is one of the traditional chinese medicinal herbs.Ootheca mantidis can be used to treat many kinds of diseases,such as deficiency of the kidney,spenmatorrhea,impotence,frequent urination,enuresis.Ootheca mantidis extracts have strong anti-oxidant activities,anti-atherosclerotic activities,anti-bacterial activities and have a good therapeutic effect on diabetic mice.Looking at related literatures at home and abroad,there are few studies on Heipiaoxiao,and there are no reports on their biological activities and chemical compositions.Objective: The biological activities of anti-oxidant,anti-bacterial,inhibitory enzyme activities,therapeutic effects on diabetes and Alzheimer's disease of different solvent extracts of Heipiaoxiao were studied,and their chemical constituents were explored.Methods:(1)Heipiaoxiao was extracted with distilled water,95% ethanol,70% ethanol,50% ethanol,n-butanol,ethyl acetate,dichloromethane and petroleum ether,respectively.Another sample of Heipiaoxiao was extracted by water extraction and ethanol precipitation,and crude polysaccharides were extracted by sevage after protein removal.A total of 9 extracts were obtained.The anti-oxidant,anti-microbial,inhibition of ?-glucosidase,?-amylase and cholinesterase activities of extracts from Heipiaoxiao in different solvents were also studied.Solvent extracts with dual inhibitory effects on acetylcholinesterase(AchE)and butyrylcholinesterase(BuChE)were studied in zebrafish with Alzheimer's disease(AD)in vivo.(2)The crude polysaccharide was obtained by hot water extration and ethanol deposition,and the polysaccharide was purified using DEAE Sephadex A-25 and Sephadex G-75 column chromatography.The polysaccharide was elucidated by GPC,HPLC and UV.In addition,the antioxidant,antibacterial,cholinesterase and acute toxicity test in mice inhibitory activity of polysaccharides were explored.(3)Separation,purification and structure identification of small molecule monomers from Heipiaoxiao by traditional extraction and separation techniques.Results:(1)70% alcohol extract and crude polysaccharide of Heipiaoxiao have good antioxidant activity,n-butanol,ethyl acetate and dichloromethane extracts all showed strong antimicrobial activity and broad antimicrobial spectrum.Crude polysaccharides inhibited a-glucosidase and n-butanol extracts inhibited a-amylase activity are strongly.The extracts of n-butanol and ethyl acetate had strong inhibitory effects on the activities of AchE and BuChE,and n-butanol extract has a significant effect on the movement disorders of zebrafish juvenile Alzheimer's disease.(2)Obtained a new polysaccharide(HPP-1)from Heipiaoxiao,the average apparent molecular weight was 13048 Da.HPP-1 have no nucleic aicd,protein and pigment.It has good anti-microbial and anti-oxidant activities,inhibits the activity of cholinesterase,and is basically non-toxic to mice.(3)11-(stearoyloxy)octadeca-9,12,15-trienoic acid(?),di-n-butyl phthalate(?)and palmitic acid(?)were isolated and purified from cuttlebone,and all of them were isolated from Heipiaoxiao for the first time.Conclusions: Crude polysaccharide,70% alcohol,n-butanol,ethyl acetate and dichloromethane extracts have better antioxidant and antibacterial activity,which has become a new direction for the development of new drugs,functional health products and green food additives.Different solvent extracts also inhibit the activities of a-glucosidase and a-amylase,and have potential in the treatment of diabetes mellitus and cardiovascular diseases.The n-butanol extract from Heipiaoxiao showed good inhibitory activity against AchE and BuChE.The zebrafish AD's experiment also showed that the n-butanol extract had better activity.The results provide a theoretical basis on research and development of double cholinesterase inhibitors and new drugs for delaying AD progression.The chemical constituents of Heipiaoxiao were studied for the first time.A polysaccharide and three small molecule monomers were obtained,which provided a reference for the basic research of Heipiaoxiao.