| Xanthium Fructus is commonly used in clinical medicine,mainly used in the treatment of rhinitis,rheumatism and other diseases.According to tradition,Xanthium "has small poison",which needs to be processed before being used as medicine.Due to the development of mechanical industry,the process of mechanical removal of Xanthium Fructus has been relatively mature.Xanthium Fructus are convenient for transportation,storage and market circulation after removal of Xanthium Fructus.At present,raw products on the market are mainly products after removal of Xanthium Fructus.In the early stage,the research group conducted a preliminary study on the sand-frying method of Xanthium after thorn removal,but it was not systematic and in-depth enough.This topic mainly optimized the sand-blanking process of Xanthium after thorn removal,and on this basis,studied the transformation rule of toxic components and toxic mechanism of Xanthium after thorn removal,in order to provide experimental basis for the formulation of technological parameters of Xanthium stir-frying method and the mechanism of toxicity reduction.Objective This project aims to optimize the sand-frying process of Xanthium Fructus from the perspective of the change of toxic component content,explore the change rule of the toxic component carboxylatricin content through the simulation processing method,and conduct a preliminary study on the mechanism of processing attenuating toxin based on HMGB1 inflammatory pathway,providing experimental basis for the processing process and attenuating mechanism of Xanthium Xanthii.Method1.Box-Behnken Design-Response surface Method optimization of Xanthium seed sanding processThe Box-Behnken design-response surface method was used to evaluate the roastingtime,roasting temperature and sand weight,and the comprehensive scores of the contents of total phenolic acid,chlorogenic acid,cryptochlorogenic acid,3,4-dicaffeoyl quinic acid,4,5-dicaffeoyl quinic acid and toxic components carboxylic atractyloside and atractyloside were used as evaluation indexes.Optimize the sanding process of Xanthium seeds.Response surface experiment design was used to optimize the processing technology of Xanthium xanthii by taking roasting time,roasting temperature and sand weight as factors,and taking the comprehensive scores of active components total phenolic acid,chlorogenic acid,cryptochlorogenic acid,3,4-dicaffeoyl quinic acid,4,5-dicaffeoyl quinic acid and toxic components carboxyl atractyloside and atractyloside as evaluation indexes.2.Study on transformation rule of toxic components of Xanthium Fructus based on simulated processing methodUPLC method was used to determine the content of carboxyl atractyloside and atractyloside in cannon products through single factor investigation of time and temperature,and to clarify the change law of the two components in cannon products.Then the carboxyl atractyloside was simulated to determine the effect of time and temperature on its transformation and to explore its transformation mechanism.3.Study on the mechanism of hepatotoxicity of Xanthium Fructus based on HMGB1 inflammatory pathwayAfter 14 days of administration,compared with normal group,body weight decreased and liver index increased in Xanthium fructus and Xanthium fructus groups(P<0.01);Serum AST,ALT,HMGB1,IL-1β,IL-2 and TNF-α levels were increased compared with normal group(P<0.01);However,the value of stir-fried product was lower than that of raw product group,with statistical difference,P<0.01;The results of WB and RT-q PCR showed that the m RNA protein expressions of HMGB1,TLR4 and NF-κB in the liver tissue of the administration group were significantly increased,but the rawproduct was more obvious than the fried product,and the difference was statistically significant.0.01,indicating that the sanding had obvious attenuating effect.Results 1.Box-Behnken Design-Response surface Method optimization of Xanthium seed sanding processAccording to the Box Behnken response surface experiment design method,roasting time,roasting temperature and sand dosage were considered as factors,and the contents of effective components chlorogenic acid,cryptochlorogenic acid,3,4-dicaffeoyl quinic acid,4,5-dicaffeoyl quinic acid,total phenolic acid and toxic components carboxylic atractyloside and atractyloside were comprehensively scored.The optimum sanding process is as follows: roasting temperature 160 ℃,roasting time 7min and sand consumption 10 times.2.Study on the transformation law of stir-fried products and simulated gun products of Xanthium seedThe results of single factor study showed that with the increase of stir-frying temperature and time,the content of carboxyl atractyloside decreased,while the content of atractyloside increased.The simulation results showed that the carboxyl atractyloside showed a similar change pattern,and the transformation amount was basically stable when the optimum processing technology was 160 degrees Celsius and 7 minutes,which verified the rationality of the sanding process parameters.3.Study on the mechanism of hepatotoxicity of Xanthium fructus based on HMGB1 inflammatory pathwayAfter 14 days of administration,compared with normal group,body weight decreased and liver index increased in Xanthium fructus and Xanthium fructus groups(P<0.01);Serum AST,ALT,HMGB1,IL-1β,IL-2 and TNF-α levels were increased compared with normal group(P<0.01);However,the value of stir-fried product was lower thanthat of raw product group,with statistical difference,P<0.01;The results of WB and RT-q PCR showed that the m RNA protein expressions of HMGB1,TLR4 and NF-κB in the liver tissue of the administration group were significantly increased,but the raw product was more obvious than the fried product,and the difference was statistically significant.0.01,indicating that the sanding had obvious attenuating effect.Conclusion The optimized processing process of Xanthium fructus was stable and feasible.The simulation processing method basically clarified the change rule of the toxic component of Xanthium Fructus carboxyl atractyloside with processing time and temperature.The results of animal experiments showed that the attenuating of Xanthium Fructus after processing may be related to the regulation of HMGB1-TLR4/(NF-κB)inflammatory pathway.This study provides preliminary experimental basis for processing technology and attenuated mechanism of Xanthium fructus. |
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