| The fruit of Xanthium sibiricum is the dried and mature fruit with involucre of Xanthium sibiricum Patr.,a plant of the composite family.Its main functions are to disperse the cold,clear the nose,dispel the wind and relieve pain.Xanthium sibiricum is "poisonous".According to the records of this herb,its processing method is mostly to fry and remove the thorn.However,with the development of mechanization,there has been a special equipment for deburring Xanthium sibiricum,which has good deburring effect and can maintain the integrity of the fruit.As a result,most of the original medicinal materials of Xanthium sibiricum in the current market have been debunked,but the processing method of Xanthium sibiricum contained in the Chinese Pharmacopoeia 2020 is still stir-fried and debunked,which has been seriously divorced from the actual situation.When the prickle-free Xanthium sibiricum is processed by the method of stir-frying,the processing end point is judged by the human eye,which is easy to lead to different appearance and properties of the finished product,and seriously affect the quality and clinical efficacy of the prepared pieces.The research group has studied the processing technology of Xanthium sibiricum by sand frying method in the early stage,but the processing effect of sand frying method and clear frying method needs further comparative study.Objective:The purpose of this study is to compare the chemical composition changes and toxicity reduction effects of the processed Xanthium sibiricum with that of the sand fried method,and explore the mechanism of toxicity reduction,so as to provide scientific basis for the rationality of the sand fried method in the processing of Xanthium sibiricum.Methods1.Fingerprint of different processed Xanthium sibiricum and determination of phenolic acids and atractylosideUPLC was used to establish the fingerprint of 45 batches of Xanthium sibiricum raw,stir-fried and sand-fried products,and cluster analysis(HCA),principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were used to compare the different components of different processed products;The contents of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,3,4-dicaffeoyl quinic acid,4,5-dicaffeoyl quinic acid,3,5-dicaffeoyl quinic acid,carboxyl atractyloside and atractyloside in different processed products were determined by UPLC.2."Dose-Time-Toxicity" Study on Liver and Kidney Toxicity of Different Processed Xanthium sibiricum240 mice were divided into 10 groups,namely,the low,medium and high groups of Xanthium sibiricum raw products,the low,medium and high groups of clear fried products,the low,medium and high groups of sand fried products and the normal group,with 24 mice in each group.Mice were gavaged with low,medium and high doses of water decoction of different processed Xanthium sibiricum seeds for 14 days,and then gavaged for 1 hour at the end of the 1st,7th and 14th days respectively.Blood was taken from 8 mice in each group.The AST,ALT,BUN and CR values in serum were measured by ELISA.At the same time,the liver and kidney tissues were weighed to calculate the organ index,and the degree of organ damage was observed after HE staining.3.Effect of different processed Xanthium sibiricum on liver drug enzyme activity in miceThirty-two mice were divided into four groups,namely,normal group,raw group,stir-frying group,and sand-frying group,with eight mice in each group.The decoction of the corresponding processed products was gavaged respectively for 14 days.The liver tissue was taken after the last gavage for 1 hour.The liver microsomes were prepared by calcium precipitation method.The mouse liver microsomes were incubated in vitro.Caffeine,tolbutamide,omeprazole,and midazolam were used as probe drugs for four enzymes CYP1A2,CYP2C9,CYP2C19,and CYP3A4,respectively,UPLC detected their consumption and calculated the relative activity of enzymes.Results1.Fingerprint of different processed Xanthium sibiricum and determination of phenolic acids and atractyloside① The fingerprints of 15 batches of raw products,stir-fried products and sandfried products were established,and 15 common peaks were calibrated.The retention time of each common peak was relatively small,and the relative peak area of different processed products was significantly different.Cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis could classify raw and stir-fried products into one category,and sand-fried products into another category,indicating that the quality of different processed products of Xanthium sibiricum was significantly different.②The contents of neochlorogenic acid,cryptochlorogenic acid,3,4-dicaffeoyl quinic acid and 4,5-dicaffeoyl quinic acid in fried and sand fried products were higher than those in raw products;The contents of chlorogenic acid,caffeic acid and 3,5-dicaffeoyl quinic acid were lower than those of raw products.③ The content of carboxyl atractyloside in fried and sand fried products decreased,while the content of atractyloside increased.2."Dose-Time-Toxicity" Study on Liver and Kidney Toxicity of Different Processed Xanthium sibiricumCompared with raw products,fried and sand fried products can reduce the ALT,AST,BUN and CR values in the serum of mice to varying degrees,improve the atrophy of liver nuclei,inflammation and congestion,and reduce the expansion,necrosis and inflammatory infiltration of renal tubular epithelial cells,showing a certain "dose-time-toxicity" relationship,and sand fried products have more obvious effect on reducing toxicity than fried products.3.Effect of different processed Xanthium sibiricum on liver drug enzyme activity in miceThe in vitro incubation system of mouse liver microsome was established.The enzyme activities of CYP1A2 and CYP3A4 in liver microsome of raw group,clear stir frying group and sand stir frying group were higher than those of normal group,with statistical difference(P<0.05).The enzyme activities of clear stir frying group and sand stir frying group were lower than those of raw group;raw products,stir fried products,and sand fried products all have varying degrees of influence on the activity of CYP2C19 and CYP2C9 enzymes in mice,but the results showed no statistical difference(P>0.05).The detoxification mechanism of processed Xanthium sibiricum may be related to changing the enzyme activity.ConclusionThe quality of sand fried Xanthium sibiricum is more uniform than that of raw and clear fried Xanthium sibiricum.After different processing,the content of the toxic component carboxyl atractyloside in Xanthium sibiricum is reduced;Animal experiments showed that the liver and kidney serum indicators and pathological damage of mice caused by stir frying Xanthium sibiricum and sand frying Xanthium sibiricum were lower than those caused by raw Xanthium sibiricum;clear fried Xanthium sibiricum and sand fried Xanthium sibiricum reduce the activity of CYP1A2 and CYP3A4 enzymes in mice,which may be the internal mechanism of their processing and detoxification.The research results indicate that both the clear frying method and the sand frying method can effectively reduce the toxicity of Xanthium sibiricum,and the sand frying method has certain advantages in processing Xanthium sibiricum. |
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