Study On The Material Basis And Toxic Effects Of Raw And Different Processed Fructus Psoraleae

Objective:According to different processing methods of Fructus Psoraleae in ancient and modern times,UPLC-Q-TOF-MS/MS technology was used to compare the differences of material basis of Fructus Psoraleae before and after processing.Based on the research foundation of our research group,the characteristics of hepatotoxicity of raw Fructus Psoraleae and water extracts of different processed products were investigated with the FXR/PPARαsignal pathway related to liver bile acid and lipid metabolism as the breakthrough point,in order to clarify the molecular mechanism of cholestatic liver injury caused by raw Fructus Psoraleae and different processed products,which is expected to prevent and treat hepatotoxicity of Fructus Psoraleae and make rational use in clinic.Methods:1.Using UPLC-Q-TOF-MS/MS technology,the components of raw and processed Fructus Psoraleae were qualitatively identified and analyzed;2.Acute toxicity test of raw psoralea corylifolia and different processed products:Kunming mice were divided into three groups:raw psoralea corylifolia group,stir-fried group,salt-roasted group,Leigong group and wine-soaked group,and blank control group,with a total of 35groups.Each group was given 0.4 ml/10g oral gavage to mice,and the observation was resumed for 14 days.The body weight,serum biochemistry and mortality were detected,and the LD₅₀value was calculated.At the same time,the liver tissue of mice was observed by HE staining.3.Taking the key targets in bile acid metabolism signal pathway and lipid metabolism pathway as the research object,the expression of target gene m RNA in bile acid metabolism signal pathway and lipid metabolism pathway was detected by q PCR method,and its toxic mechanism was preliminarily explored.Results:1.After processing by different processing methods,the content of psoralen,a toxic component reported in the literature,was significantly lower in Leigong group,wine-soaked group and salt-roasted group compared with raw psoralen and stir-fried group,among which Leigong method could reduce the content of some potential toxic components and increase the content of some active components with liver protection and anti-inflammation;2.There was no statistical difference in the weight of mice before different methods of administration.On the first day of administration,the weight of mice in the administration group decreased to some extent（except for the females in the raw product group and some dosage groups in other groups）,and the overall weight of mice increased from the fourth day,but the weight of mice in each administration group was basically smaller than that of mice in the control group;The LD₅₀values of crude drug group,stir-fried group,salt-roasted group,tripterygium wilfordii group and wine-soaked group were 63.20,56.92,51.95,88.61 and 59.02 g crude drug/kg,respectively,indicating that tripterygium wilfordii group showed the smallest acute toxicity.Compared with the control group,male mice:the serum ALT and TBA levels in the raw group increased significantly;The serum ALT,AST and TBA in the stir-fried group increased significantly,but the ALP level was not statistically significant.The levels of serum ALP,ALT,AST and TBA in salt-roasted group,Leigong group and liquor-soaked group increased significantly.Female mice:the serum ALP,ALT,AST and TBA levels in the raw group,salt-roasted group,Leigong group and wine-soaked group increased significantly;Serum ALT,AST and TBA levels in the stir-fried group increased significantly;TP levels in all groups decreased significantly.Pathological results showed that there was no liver injury in the control group.Pathological changes such as vacuolar degeneration of hepatocytes,glycogen degeneration of hepatocytes,cytoplasm looseness of hepatocytes,etc.occurred in the livers of mice in the raw group,stir-fried group,salt-roasted group,Leigong group and wine-soaked group.The severity of liver pathological changes caused by different processing methods:raw food group>wine soaking group>stir-frying group>salt roasting group>Leigong group,in which the liver pathological changes caused by wine soaking method are relatively serious,while the liver pathological changes caused by Leigong method are the lightest;The mechanism study found that compared with the blank group,the expression of gene FXR in male mice:salt-baked group was significantly reduced;The expression of PPARαgene in raw product group,stir-fried group and Leigong group decreased significantly;Compared with the raw product group,the FXR of the wine-soaked group was significantly lower,but the FXR of the Leigong group did not change significantly.Female mice:compared with the blank group,the expressions of FXR and PPARαin the salt-baked group and the wine-soaked group were significantly lower;Compared with the raw product group,the PPARαof mice in the wine immersion group was significantly lower.Conclusion:Psoralea corylifolia can reduce the content of psoralen,isopsoralen and other main toxic components by processing methods such as Leigong,wine soaking and salt roasting.Combined with the LD₅₀value,Leigong’s 88.61 g crude drug/kg is 521 times the daily dosage of adults per kilogram of body weight（about 1.5 times that of other groups）,and Leigong group has the most obvious attenuation effect after processing.According to the changes of ALT,AST,TBA in hematology,liver pathology,and the expression results of related genes,such as FXR,a regulator of bile acid in liver,and PPAR signal pathway related to lipid metabolism,it was found that Psoralea corylifolia L.was still toxic to some extent after being processed by different processing methods,but the toxicity of Psoralea corylifolia L.was weaker than that of the raw product group,and the effect of Leigong method on FXR/PPARαsignal pathway related to the reported toxicity of Psoralea corylifolia L.was the least.