Studies On Metabolites And Mechanism Of "Sweating" Salvia Miltiorrhiza Bge. Based On "Color Discrimination Grading"

Salvia miltiorrhiza.is the dried root and rhizome of Salvia miltiorrhiza Bge.which was first recorded in shennong herbal scripture.It is bitter and slightly cold.Heart,liver meridian;it has the functions of promoting blood circulation and removing blood stasis,relieving pain through menstruation,clearing heart and removing annoyance,cooling blood and eliminating carbuncle.It is mainly used for abdominal and hypochondriac pain,abdominal mass accumulation,heat arthralgia pain,upset and insomnia,irregular menstruation,dysmenorrhea and amenorrhea,sores and swelling pain and other diseases.In the departmental standard,Danshen has a record of“sweating”.Practice has proved that“sweating Danshen”has excellent quality and excellent quality.However,the“sweating”method and other factors have a certain impact on the effective components of Salvia miltiorrhiza,resulting in uneven quality of medicinal materials.Objective:Taking“mild sweating”,“severe sweating”and“non-sweating”Salvia miltiorrhiza as the research objects,the changes of active components in different tissue parts of Salvia miltiorrhiza were compared to explore whether the active enzyme catalyzed the transformation of active components in the cork,cortex and xylem during the“sweating”process of Salvia miltiorrhiza,resulting in changes in the color of each tissue part,and ultimately affecting the changes of pharmacological activity of Salvia miltiorrhiza.To explore whether“sweating”treatment enhances the pharmacological effects and material basis of Salvia miltiorrhiza.Try to provide an explanation for the mechanism of the traditional processing method of Salvia miltiorrhiza“sweating”.Methods:Laser capture microdissection(LMD)technology,colorimeter method,enzyme-linked immunosorbent assay,UPLC-Q-Exactive-MS/MS,liquid chromatography,DPPH and ABTS free radical scavenging experiments were used to study the angiogenic activity of different tissues of’sweating’and’non-sweating’Salvia miltiorrhiza.On this basis,combined with multivariate statistical analysis technology,the correlation analysis of color quantitative index-enzyme activity-main differential metabolites-pharmacological activity in the process of’sweating’of Salvia miltiorrhiza was conducted.Results:The color index of Salvia miltiorrhiza after’sweating’treatment changed significantly.The deeper the degree of’sweating’was,the smaller the lightness value(ΔL~*)was.The red-green value(Δa~*)of the cork layer of non-sweating Salvia miltiorrhiza was larger than that of mild and severe sweating Salvia miltiorrhiza.The red-green value(Δa~*)and yellow-blue value(Δb~*)of the cortex xylem of mild and severe sweating Salvia miltiorrhiza were larger than those of non-sweating Salvia miltiorrhiza.The tyrosinase content and PPO activity of various parts of’sweating’Salvia miltiorrhiza samples increased with the deepening of’sweating’,and the POD activity decreased with the deepening of’sweating’.A total of 84 differential compounds were screened before and after’sweating’of Salvia miltiorrhiza.There were 36 differential metabolites(L-tyrosine,adenine,lithospermic acid,chlorogenic acid,salvianolic acid A,danshensu,rosmarinic acid,salvianolic acid C,etc.)increased,and 39 differential metabolites(L-asparagine,2’-deoxyguanosine,tanshinone IIA,isoferulic acid,protocatechuic aldehyde,etc.)decreased.In addition,four metabolites(adenosine,adenosine 3’-monophosphate,feruloyl glucose,caffeic acid)decreased in mild sweating and increased in severe sweating.There were five metabolites(cytidine,cytidine-5’-monophosphate,salvianolic acid I,salvianolic acid K,epicatechin gallate)increased in mild sweating and decreased in severe sweating.Five metabolic pathways were found to be closely related to the mechanism of’sweating’,including amino acid synthesis,amino acid metabolism,carboxylic acid metabolism,glucose metabolism and plant hormone signal transduction.The order of DPPH free radical scavenging ability of 9 parts of Salvia miltiorrhiza was non-sweating cork>mild sweating cork>severe sweating cork>severe sweating cortex>mild sweating cortex>severe sweating xylem>mild sweating xylem>non-sweating xylem>non-sweating cortex.The order of ABTS free radical scavenging ability was mild sweating cork>non-sweating cork>severe sweating cork>mild sweating xylem>severe sweating cortex>mild sweating cortex>severe sweating xylem>non-sweating cortex>non-sweating xylem.All parts of Salvia miltiorrhiza have different degrees of promoting effect on zebrafish angiogenesis,and the overall promoting effect is cortex>xylem>cork.The cortex and cork cortex of Salvia miltiorrhiza after sweating are more effective than the same parts of Salvia miltiorrhiza without sweating in promoting zebrafish angiogenesis.Conclusion:The brightness value(ΔL~*)of Salvia miltiorrhiza after’sweating’treatment became smaller and the color of medicinal materials became darker.The results of correlation analysis showed that the content of ester-soluble components of Salvia miltiorrhiza was significantly correlated with the red-green value(Δa~*).To cope with the’sweating’stress process,Salvia miltiorrhiza metabolizes acid substances and increases tyrosine content.’Sweating’is the condition to promote the transformation and redistribution of effective components of Salvia miltiorrhiza.The change and distribution of components affect the pharmacodynamic activity of Salvia miltiorrhiza.’Sweating’generally enhanced the antioxidant activity and promoted new angiogenesis activity of Salvia miltiorrhiza.The cork layer and cortex are the most obvious parts of the color change of’Fahan’Salvia miltiorrhiza,and the pharmacodynamic activity is the strongest.At the same time,’sweating’improved the resource utilization of the inner tissue of Danshen suppository.Therefore,’sweating’can be used as a processing method to improve the quality of Salvia miltiorrhiza is scientific and reliable,and its cost is less worthy of promotion.