Investigation On The Processing Technology Of Wine-Processed Fructus Corni And Its Synergistic Mechanism

In this paper,the processing technology of Fructus Corni was strictly screened at first,and the important factors in the processing technology,such as steaming time,steaming temperature,moistening time,and the amount of alcohol added were determined.Meanwhile,the protective effectsof Fructus Corni on the human renal cortex proximal convoluted tubular epithelial cell(HK-2)and rat hepatic stellate cell(HSC-T6)of the fibrosis disease model were innovatively selected as the indices.The levels of each factor were determined through single-factor investigation,and the best processing conditions for different processed products,such as steaming,high-pressure steaming,wine steaming,high-pressure wine steaming,wine immersion,and wine stirfrying were obtained by orthogonal test.Finally,the stability and repeatability of the processing technology were verified.After determination of the processing technology of six processed products,including steaming,high-pressure steaming,wine steaming,high-pressure wine steaming,wine immersion,and wine stir-frying,the secondary metabolites in crude and wine-processed Fructus Corni were comprehensively described by UHPLC-Q-TOFMS/MS,and the changes of carbohydrate composition in crude and wine-processed Fructus Corni were analyzed by multidimensional chromatography with HPLC(C18)UV,HPLC(NH2)-ELSD,and HPGPC-ELSD.At the same time,the above described two methods were combined to find differential substances for simulated processing,and then the chemical transformation mechanism of Fructus Corni processing was elucidated through simulated processing.The connotation of the processing technology of Fructus Corni was further clarified from the perspective of the whole composition of Fructus Corni.According to the protective effects of crude and processed Fructus Corni on the fibrosis levels of HK-2 and HSC-T6 cells in vitro and the changes of chemical composition,high-pressure wine steaming was determined as the best processing technology of Fructus Corni.The rat model of hepatic fibrosis was replicated by subcutaneous injection of 40%carbon tetrachloride(CCl4)olive oil solution,and the model rats were intragastrically administered with the decoctions of crude and wineprocessed Fructus Corni.After eight weeks,all the rats administered with the decoctions of crude and wine-processed Fructus Corni were tested to have the decreased levels(P<0.05)of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and liver hydroxyprolinase(Hyp).The results of HE and Masson staining showed that the inflammatory infiltration of liver was reduced and the fibroplasia was improved.The protein expression levels of α-smooth muscle actin protein(α-SMA)and collagen Ⅰ were detected by Western-blot and immunofluorescence,and the results showed that their protein expression levels were all decreased(P<0.05).Taking all the indices into consideration,the anti-hepatic fibrosis effect in rats of wine-processed Fructus Corni was better than that of crude one,and the improvement effect of high-dose was the best.Based on the results of pharmacodynamic investigation,the plasma metabolic profiles of the blank control group,the model group,and the high-dose groups of crude and wine-processed Fructus Corni were analyzed by UHPLC-Q-TOF-MS/MS technology.The results showed that the plasma samples of the model group and the blank control group could be clearly divided into two types.The groups of crude and wine-processed Fructus Corni were separated from the model group and moved to the blank control group after intragastric administration of the decoctions of crude and wine-processed Fructus Corni,suggesting that crude and wine-processed Fructus Corni could interfere with hepatic fibrosis in the model group and restore the metabolic disorders of the body.Compared to the blank control group,the twenty-four potential biomarkers related to hepatic fibrosis were screened and identified in the model group,which mainly involved in primary bile acid metabolism,glycerol phospholipid metabolism,pentose and glucuronide metabolism,retinol metabolism,and arachidonic acid metabolism.The decoctions of crude and wine-processed Fructus Corni had different degrees of callback effects on the ten of the above potential biomarkers,and the callback effect of wine-processed Fructus Corni was closer to the blank control group,and better than that of crude one.The present study clarifies the mechanism of enhanced efficiency of wine-processed Fructus Corni from the perspective of plasma metabolism,and provides the theoretical foundation for further development and clinical application of Fructus Corni.