| Objective Using poria cocos and millet rice as raw materials,a health care rice wine with pure color,fragrance and good taste was developed,and its function was studied,which provided a new idea for the study of health care rice wine in treating spleen Yang deficiency and improving body immunity.Methods 1.Taking reducing sugar content as index,the saccharification temperature,saccharification amount of wheat koji and saccharification time were selected to investigate the saccharification process of traditional poria cocos wine by single factor.Combined with the results of single factor investigation,L9(3~4)orthogonal test was designed to obtain the best saccharification process.The fermentation process of traditional poria cocos wine was investigated by single factor,including yeast inoculation amount,fermentation days and inoculation temperature,and the optimal fermentation process was obtained by L9(3~4)orthogonal test combined with the results of single factor investigation.2.The main functional components of traditional poria cocos wine were analyzed and their contents were determined by ultraviolet spectrophotometry(UV-Vis),and the contents of poria acid,catechin and ferulic acid in traditional poria cocos wine were determined by high performance liquid chromatography(HPLC).The types and relative contents of volatile flavor components in traditional poria cocos wine were determined by gas chromatography and mass spectrometry(GC-MS).According to GB/T 13662-2018"Yellow Rice Wine"national standard,the quality of traditional poria cocos wine was evaluated.3.The spleen-yang deficiency model of rats was established by three factors,including diet insufficiency,excessive tiredness and bitter cold purging.The five aspects of physical signs,viscera coefficient,gastrointestinal function indexes,immune-inflammation related indexes and pathological changes of spleen were studied in the model rats.The spleen strengthening effect of traditional poria cocos wine was discussed,and the liver function indexes were determined to evaluate its influence on liver function.4.The immune hypoplasia model of mice was established by subcutaneous injection of hydrocortisone.According to the indexes of organ coefficient,carbon clearance test,delayed allergic reaction,splenic lymphocyte transformation test and T lymphocyte subsets,the immune regulation effect of traditional poria cocos wine was discussed.Results 1.Through single factor investigation and orthogonal test,the optimal saccharification process of traditional poria cocos wine was determined as adding 8%wheat koji and saccharifying at 60℃for 40 min.The optimal fermentation process was inoculated with 0.2%yeast at 28℃and fermented for 7 days in container.It has been proved that the optimized process is stable,reasonable and feasible.2.The content of main functional components in traditional poria cocos wine was determined.The results showed that polysaccharide content was 70.69±1.44g/L,which was 1.67 times that of traditional millet rice wine.The contents of poria acid,catechin and ferulic acid were 0.27±0.01 mg/L,47.66±1.11 mg/L and 2.27±0.03 mg/L,respectively.Forty volatile components were detected in traditional poria cocos wine,among which the contents of alcohol and aromatic substances were the most,which were 42.32%and 47.36%of the relative contents,respectively.The quality evaluation of traditional poria cocos wine was carried out,and the results met the national standard of GB/T 13662-2018"Yellow Rice Wine".The physical and chemical indexes were:total sugar 78.1 g/L,non-sugar solid 36.6 g/L,alcohol content12.0%vol,p H 4.18,total acid 4.11 g/L,amino acid nitrogen 0.18 g/L.Calcium oxide 0.4g/L.3.In the rat model with spleen-yang deficiency,high dose of traditional poria cocos wine could promote the increase of body weight,improve the lesion degree of spleen,and significantly increase serum D-xylose content(P<0.01)and serum interferonγ(IFN-γ)content(P<0.05).It has certain therapeutic effect on digestion,absorption and immune regulation of spleen-yang deficiency rats.In addition to the above effects,traditional poria cocos wine in medium dose can also significantly reduce the contents of serum gastrin(GAS),interleukin-6(IL-6)and monocyte chemotactic protein-1(MCP-1)(P<0.05),better improve the digestion and absorption function of rats with spleen-yang deficiency and play a role in immune regulation.In addition,compared with the poria macerating wine prepared from distilled wine,the serum alkaline phosphatase(ALP)level and serum alanine aminotransferase(ALT)level were not significantly increased at medium dose of traditional poria cocos wine,indicating that traditional poria cocos wine had no obvious liver toxicity.4.In the immunocompromised mouse model,the immune enhancement effect was most significant in traditional poria cocos wine high-dose group,which could significantly increase the spleen index of mice(P<0.01),increase the degree of ear swelling of mice(P<0.05),and improve the proliferation ability of splenic lymphocytes of mice(P<0.05).Down-regulated the percentage of CD3~+CD8~+cells(P<0.01),up-regulated the percentage of CD3~+CD4~+cells(P<0.01)and the ratio of CD4~+/CD8~+cells(P<0.01),indicating that traditional poria cocos wine may improve the immune function of the body by regulating cellular immunity.Conclusion The preparation process of traditional poria cocos wine optimized in this study is reasonable,feasible and highly workable.The finished wine has excellent taste,pure color and flavor,and rich nutrition,which conforms to the GB/T 13662-2018"Yellow Rice Wine"national standard.It can effectively improve the gastrointestinal dysfunction caused by the deficiency of spleen Yang,enhance the immunity of the body,play the function of invigorating the spleen and stomach.This study provides a theoretical basis for the study of health rice wine in treating spleen-yang deficiency and improving body immunity,and broadens the idea for further development and utilization of poria cocos,a traditional Chinese medicine and food. |
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