| Studies on toxicity and carcinogenicity of areca nut began in the west and India,where people used to eat it with betel piper and slaked lime.People in mainland China,especially Hunan province,usually eat the dried areca nut,which is made by a series of processing steps,such as cooking and ordering brine.Polyphenols and arecoline components of these treated areca nut product were greatly reduced.Besides,with the widespread use of smokeless cold fumigation process,green areca nut has gradually occupied the market in recent years.The rise of microbiomics and metabolomics provides a better way to study the relationship between areca nut consumption and disease than before.In this experiment,the aqueous solution of areca was extracted by steps of freezing crushing,ultrasound,water bath and centrifugation,and the contents of arecoline and arecaidine were analyzed by liquid chromatography.On this basis,different gavage solutions were prepared and given to mice.The colon and serum samples of mice treated with different gavage solutions were analyzed by 16 S metagenomics and untargeted metabolomics,and the possible relationship between the consumption of processed areca nut and disease was preliminarily discussed.The main results were as followed:1.The content of arecoline in the water extract of fresh areca,green areca without brine,smoked areca and green areca nut with brine were 2.71±0.15mg/m L,1.76±0.01mg/m L,1.79±0.01mg/m L and 1.30±0.01mg/m L.And,arecaidine in different water extract were0.78±0.05mg/m L,0.42±0.01mg/m L,0.50±0.06mg/m L and 0.76±0.01mg/m L,respectively.The content of arecoline was higher than arecaidine in all four kinds of areca water extracts.The content of arecoline and arecaidine in green areca and smoked areca nut were significantly lower than that in fresh areca,which suffered great loss during production.2.Mice of different groups were given corresponding gavage solution every day and whole blood was taken and analyzed after 28 days.The results showed that pure arecoline and water extracts of different areca nut did not cause significant differences in body weight,food intake and blood routine indexes of these mice compared with the control group.3.The colonic contents of each group of mice were collected after 28 days of gavage,and the colonic flora before and after gavage was analyzed by 16 S metagenomic sequencing.The results showed that the abundance of lactobacillus in mice colon was significantly changed after four weeks of gavage.The colonic flora of mice gavage smoked areca and fresh areca water extract were significantly different at multi-taxonomic levels,include Bacteroidetes,Firmicutes,Tenericutes,Alloprevotella and Parabacteroides distasonis.This suggests that the metabolism of carbohydrate in their colon may be different.4.Serum from different groups of mice was collected and metabolites were identified and quantified by high resolution liquid and mass spectrometry after four weeks of gavage trial.The results showed that the intragastric administration of smoked areca,green areca nut extract and arecoline could significantly increase the bioactive serum metabolites such as 2-Amino-1,3,4-octadecanetriol,cuminaldehyde,apocynin,trans-cinnamaldehyde,α-ergocryptine and hydrocinnamic acid.Results also showed that the serum content of metabolites such as progesterone,guggulsterone,vitamin C and naringenin was significantly lower in arecoline treated mice than the smoked areca nut extract treated ones.In addition,the content of various lipid metabolites also changed significantly.In summary,smoked areca,green areca nut extracts and arecoline may have certain influence on the occurrence of disease of normal mice under the dosage of this experiment,and the changes of a series of bioactive metabolites is closely associated with the arecoline component in the areca nut solution. |
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