Analysis Of The Chemical Compositions And Antitumor Activities Of Water Extracts From Walnut Diaphragm

Diaphragma juglandis Fructus is the dry wood diaphragm within kernel from walnut of juglandaceae plants,is a traditional Chinese medicinal materials,The dictionary of Chinese medicine has recorded that it was mainly used in the treatment of kidney empty,urinary frequency and diarrhea,modern medical research has shown that wood has pharmacological functions of bacteriostatic,kidney,antioxidant and antitumor.In daily life,people often used to make tea to drink.Now,the study about chemical composition and physiological activity of water extracts from Walnut Diaphragm was rarely reported.Based on distracted by water extracts from Walnut Diaphragm as the research object,studied its volatile components in the system,established a new method to analysis Chemical composition of Walnut Diaphragm,at the same time evaluated the antitumor activity of the water extraction,and used western blot to study the molecular mechanisms of water extracts from Walnut Diaphragm triggers cancer cell apoptosis.The main research contents and conclusions were as follows:(1)Analysis of the Volatile Compositions of Water Extracts from Walnut Diaphra gmThe volatile compounds of water extracts from walnut diaphragm were extracted by using four different polar solid-phase micro-extraction(SPME)fibers,separated by either of two chromatographic columns,HP-5(with weak polarity)and RTX-WAX(with strong polarity),choosed its enrichment conditions,and optimized its separation conditions.A method to analysis the volatile compounds of Walnut Diaphragm using HS-SPME-GC-MS is established.Further,the effects of different chromatographic columns and extraction fibers on the analysis of volatile components were examined.Results showed that a total of 115 volatile compounds with known structure from four walnut diaphragm extracts were separated on different chromatographic columns and identified by GC-MS,consisting of 21 alcohols,18 esters,15 ketones,14 aromatic compounds,13 hydrocarbons,12 aldehydes,6 acids,5 phenolic compounds,2 ethers and 9 other compounds.The main volatile components were acetic acid,ethyl acetate,2,6-dibutyl-4-methylphenol,menthol,cedrol,2,3-butanediol,1-hexanol,phenethylalcohol,3-butyl-4-hydroxyanisole,hexanal,and azulene.(2)Chemical composition of Water Extracts from Walnut Diaphragm identified by HPLC-MSThe kinds of difficulty volatile,high boiling point and poor heat stability compounds of Water Extracts from Walnut Diaphragm was identified by HPLC-UHD accurate mass Q-TOF MS,A new method to analysis Chemical composition of Walnut Diaphragm is established.The kinds of chemical compounds were included gallic acid,catechin and quercetin,4-Methylumbelliferyl-?-D-glucoside and ellagic acid.Quercetin,4-Methylumbelliferyl-?-D-glucoside and ellagic acid have been reported for the first time;This analysis method is simple and high sensitivity,and its result is reliable.(3)Study of antitumor activities and mechanisms of Water Extracts from Walnut DiaphragmThe ability of Water Extracts from Walnut Diaphragm inhibite HCT-116 human colon cancer cell proliferation was assessed by MTT and flow cytometry.The results showed that Water Extracts from Walnut Diaphragm could significantly inhibit proliferation of HCT116 colon cancer cell in dose dependent manner at 50~400?g/m L and IC50 value is(208.78±4.22)?g/mL.The ability of cell proliferation inhibite of Water Extracts from Walnut Diaphragm is weaker(the concentration at 50~400?g/m L)compare to the clinical first-line chemotherapy drugs 5-fluorouracil,treat cancer cell with water extracts from Walnut Diaphragm and 5-fluorouracil alone or incombination,water extracts from Walnut Diaphragm synergistically promotes 5-fluorouracil-mediated apoptosis.The molecular mechanisms of water extracts from Walnut Diaphragm triggered apoptosis was invested by western blot,results show that water extracts from Walnut Diaphragm triggers HCT-116 human colon cancer cell apoptosis through endoplasmic reticulum stress pathway.