The Study On The Antioxidant Activity Of Selenium Enriched Cultivation Of Brccoli And Its Inhibitory Effect On Human Lung Cancer Cells A549

Broccoli,an anti-cancer expert in vegetables because of its abundant sulforaphane of antitumor activity,belongs to the Brassica genus Cruciferae family.Broccoli is rich in sulfur compounds,as well as selenium,which also has biological characteristics of anticancer and anti-aging.There may be a better health care effect in terms of comprehensive utilization of both.In order to investigate the change of activity of antioxidant and antitumor after broccoli cultivated with selenium,we took selenium enriched varieties of broccoli cultivated in Enshi city,Hubei province as raw material to analyze the effect of selenium enriched cultivation on nutrition of different parts of broccoli,selenium content and sulforaphane content;extracting the broccoli curd,stems and leaves of selenium enriched broccoli and ordinary broccoli with three different polar solvent,water,75% ethanol,and ethyl acetate respectively,we got extracts of different polarity of different parts of broccoli.DPPH free radical scavenging system,hydroxyl radical scavenging system and FRAP metal ion reduction system were used to measure antioxidant activity of the extracts of different polarity;Cell culture technique in vitro and MTT assay were used to detect anti-proliferation effects on human lung cancer cells,selecting alcohol extract of relatively strong anti-cancer activity in broccoli curd to act on cells,then observing the cell morphological changes after alcohol extract treated on A549 by inverted miscroscope,the rate of apoptotic cell were respectively detected by Annexin V-FITC/PI.Results are as follows:1.There was no significant correlation between the nutrition content of broccoli and selenium enriched cultivation.Total sugar content was determined by 3,5-dinitrosalicylic acid assay,total sugar content of different parts of broccoli cultivated with selenium all increased,the highest of stems of selenium enriched broccoli was 570.31±6.52 mg/g,the total phenol content of broccoli was measured by Folin-Denis method,total flavonoids by aluminum nitrate colorimetric method,vitamin C content by 2,6-two chlorine indophenol method,total protein content by Kaumas Bradford method,and the results showed no significant correlation between the nutrition content and selenium enriched cultivation.2.Selenium enriched cultivation significantly enhanced the selenium content of broccoli.Using HG AFS to analyze total selenium content and organic selenium content,the order of selenium content of different parts was: broccoli curd>leaves>stems,the total selenium content of ordinary broccoli curd,stems and leaves was 29.05±0.83?g/g?12.82±0.63?g/g?14.65±0.24?g/g,the total selenium content of selenium enriched broccoli curd,stems and leaves was 44.85±2.4?g/g?29.40±1.21?g/g?39.82±1.22?g/g,increased by 54.39% ? 129.33% ? 171.81%;the organic selenium content of ordinary broccoli curd,stems and leaves was 21.32±0.96?g/g?9.56±0.62?g/g?11.44±0.22?g/,the organic selenium content of selenium enriched broccoli curd,stems and leaves was36.98±2.5?g/g?21.10±1.66 ?g/g?31.98±1.98 ?g/g,increased by 73.45%?120.71%?179.55% respectively.3.Selenium enriched cultivation significantly decreased the sulforaphane content of broccoli.Sulforaphane content in broccoli was determined by HPLC,the order of sulforaphane content in different parts was: broccoli curd>stems>leaves,the sulforaphane content of different parts in ordinary broccoli was 77.70±5.27?g/g?39.91±2.36?g/g?27.50±4.10?g/g,the sulforaphane content of different parts in selenium enriched broccoli was 15.30±1.34?g/g?5.19±0.67?g/g?5.08±0.95?g/g,decreased by 80.31%?86.99%?81.53% respectively.4.The antioxidant activity of enriched broccoli was stronger than ordinary broccoli,the order of the antioxidant activity of different parts of was leaves>broccoli curd>stems,the order of the antioxidant activity of three different kinds of extracts was alcohol extracts>aqueous extracts>ester extracts.Through DPPH assay,phenanthroline assay,and FRAP assay,we found that solvent extracts of different polar of selenium enriched broccoli and ordinary broccoli all had different levels of antioxidant activity,the total antioxidant activity of aqueous extracts of enriched broccoli and ordinary broccoli in broccoli curd,stems and leaves was 0.38±0.02mmol/g ? 0.26±0.01mmol/g ?0.53±0.02mmol/g?0.29±0.02mmol/g?0.12±0.03mmol/g?0.48±0.02mmol/g,the total antioxidant activity of alcohol extracts was 0.56±0.06mmol/g ? 0.26±0.02mmol/g ?0.83±0.05mmol/g?0.40±0.01mmol/g?0.22±0.02mmol/g?0.80±0.01mmol/g,the total antioxidant activity of ester extracts was 0.29±0.02mmol/g ? 0.19±0.02mmol/g ?0.34±0.01mmol/g ?0.21±0.03mmol/g?0.20±0.01mmol/g?0.25±0.02mmol/g,overall,antioxidant activity of enriched broccoli was stronger than ordinary one.5.Selenium enriched cultivation improved broccoli's anti-proliferation effects on human lung cancer cells.Only alcohol extracts in three different kinds of solvent extracts had a good inhibitory effect on human lung cancer cells A549,and also had a dose-effect;broccoli curd and leaves had a better anti-proliferation activity compared with that of the stems,which was the lowest,and anti-proliferation activity of the alcohol extracts of enriched broccoli was stronger than that of ordinary broccoli.