The Process Of Rhizoma Polygonati Fermented By Monascus And Its Immunoregulation And Hypolipidemic Effects

Objective:In this study,to improve the color and taste of Rhizoma Polygonati products,and to enhance its edible and application value.Monascus was used to solid-state fermented Rhizoma Polygonati,to screen the best technological conditions of fermentation,and to carry out digital quality control on the whole process.Compare the difference of immunoregulation and hypolipidemic function of Rhizoma Polygonati obtained by fermentation method and alcohol production method,and discuss the significance of fermentation,provide scientific basis for the application of fermentation method in the processing of Rhizoma Polygonati,and also provide a reference for the identification and quality evaluation of Monascus fermented Rhizoma Polygonati.Methods:?1?Optimization of the process of fermentation:Rhizoma Polygonati was used as the substrate,Monascus was inoculated for solid-state fermentation,and the content of lovastatin in the fermentation product was determined by HPLC.Taking the content of lovastatin as the evaluation index,the optimal content of each component of the fermentation medium was investigated by single factor test and orthogonal test;on the basis of optimizing the medium,the influence of technological parameters on fermentation was investigated by single factor test and response surface test.?2?Digital quality control of fermentation process:the chromaticity value of Monascus fermented Rhizoma Polygonati was measured by electronic sensory technique?the color chromatic aberration meter?,and the difference in chromaticity value of samples at different fermentation times was analyzed and compared.The content of polysaccharides and total saponins was determined by colorimetry.The content of lovastatin and citrinin were determined by HPLC.SPSS 20.0 and SIMCA 13.0 were used for multivariate statistical analysis,including multidimensional scaling analysis,cluster analysis,principal component analysis,correlation analysis and regression analysis,to study the relationship between color and active components in the fermentation process.?3?The immunoregulation effect of Monascus fermented Rhizoma Polygonati:The immunosuppressive model of mice was established by intraperitoneal injection of cyclophosphamide and gavage for 14 days.Serum IgG,TNF-?,IL-4 were measured by ELISA,TLR4,MyD88,TRAF-6,NF-?B mRNA expression in spleen were detected by QPCR,and TLR4,MyD88 in spleen were detected by Western blot,NF-?B protein expression level.?4?The hypolipidemic effect of Monascus fermented Rhizoma Polygonati:a high-fat mouse model was established by feeding high-fat feed,and intragastric administration was carried out.After 8 weeks,TC,TG,HDL-C,The content of LDL-C is compared with that of traditional artillery product Rhizoma Polygonati.Results:?1?Optimization of the process of fermentation:the optimal medium group was divided into fructose 6%,soybean powder 12%,KCl0.1%,KH₂PO₄ 0.4%and MgSO₄ 0.2%.The optimum fermentation parameters were as follows:500 mL wide mouth conical flask,25g of loading,60%of initial water content of medium,15%of inoculating,6%of water supply,incubate at 30°C,the temperature was reduced to 25°C on the 7th day of fermentation,and the fermentation period was 15 days.?2?Digital quality control of fermentation process:There were significant differences in chromaticity values L*,b*and E*ab among samples at different fermentation stages?P<0.05?.The content of polysaccharide and saponin increased after fermentation.With the prolongation of fermentation time,the content of polysaccharide gradually increased to the peak value on day 9 and then gradually decreased,the fermented product?d15?had a polysaccharide content of 46.21 mg/g,which was significantly different from unfermented?d0??P<0.01?.The content of saponin showed an overall rising trend,with a slight fluctuation in the middle,the fermented product?d15?had a saponin content of 8.64mg/g,which was significantly different from unfermented?d0??P<0.01?.Lovastatin levels peaked on day 15?15.39?g/g?,and have fallen sharply since then.The citrinin content of the fermentation products did not exceed 50?g/kg,which was in line with light industry standards.Multi-dimensional scale analysis,cluster analysis and principal component analysis have established discriminant models at different stages of fermentation,and divided the samples into four categories according to fermentation time:0?3 d,3?7 d,7?11 d,and 11?18 d.The content of polysaccharide was positively related to the lightness value L*and the total color value E*ab?P<0.01?,and the contents of saponin and lovastatin were significantly related to the red-green value a*?P<0.01?.The content prediction model was established through regression analysis,the saponin content can be predicted by a*and L*to the extent of 54.1%,while the lovastatin content can be predicted by a*to the extent of 61.9%.?3?The immunoregulation effect of Monascus fermented Rhizoma Polygonati:Compared with the normal group,the thymus index,and the content of Ig G,TNF-?,IL-4 in the model group was significantly reduced?P<0.01?.Compared with the model group,the levels of IgG,TNF-?and IL-4 in the serum of MRP medium and high dose groups were significantly increased?P<0.01?;the mRNA expression levels of TLR4,MyD88,TRAF-6 and NF-?B in the spleen of the MRP high-dose group were significantly increased?P<0.05?,TLR4,MyD88 and NF-?B protein expression levels were significantly increased?P<0.05?.The TLR4/NF-?B signaling pathway may be one of the related pathways for MRP to play an immunomodulatory role.?4?The hypolipidemic effect of Monascus fermented Rhizoma Polygonati:Compared with the normal group,the serum TC,TG and LDL-C levels in the model group were significantly increased?P<0.01?,and the HDL-C levels were significantly decreased?P<0.05?.Compared with the model group,the levels of TC,TG and LDL-C in the serum of mice in the middle and high dose groups of MRP decreased significantly?P<0.05?,and the levels of HDL-C increased significantly?P<0.01?.Conclusion:After Monascus fermented,the color and taste of Rhizoma Polygonati are improved,which improves the food and application value of Rhizoma Polygonati and enriches the product forms of Rhizoma Polygonati.MRP can enhance the immune function of immunocompromised mice and adjust the level of blood lipids of mice.The introduction of electronic sensory technology?the color difference meter?can provide a simple and fast inspection method for the dynamic monitoring and prediction of the active ingredient content in the fermentation process,which is conducive to the interpretation of the scientific connotation of the"distinguishing feature"of traditional Chinese medicine.