| Objective:Simulated the traditional "sweating" to deal with Salvia miltiorrhiza,the detection in the process of "sweating" in Salvia miltiorrhiza ingredients dynamic changes in the trend.Through the analysis of sweating and none Salvia miltiorrhiza and in vivo fingerprint,and antioxidant activity,protective effect of myocardial cells,preliminary comparative study on the efficacy of cerebral protective effect.Explore the effect of sweating on the quality and efficacy of Salvia miltiorrhiza.In order to provide some experimental basis for the explanation of the scientific connotation of the "sweating" method based on the traditional experience.Method:(1)The simulation of traditional "sweating" of Salvia miltiorrhiza treated with the method of "sweating" process of continuous dynamic sampling,according to the China Pharmacopoeia 2015 edition,HPLC method was used to determine the contents of cryptotanshinone,tanshinone I,tanshinone IIA and salvianolic acid B in Salvia miltiorrhiza.Analysis on the change of main index components of Salvia miltiorrhiza.(2)To establish the fingerprint of sweeting of Salvia miltiorrhiza by HPLC method,HPLC fingerprint information analysis combined with principal component analysis,comparative analysis of sweeting and none Salvia miltiorrhiza,explore the changes in the overall composition of Salvia miltiorrhiza in "sweating" in the process of looking for the similarities and differences between the overall composition of sweeting and none Salvia miltiorrhiza,for identification of sweating and none Salvia miltiorrhiza to discrimination and identification.(3)The HPLC fingerprint and the variance analysis of common peak area,research on sweeting and none Salvia miltiorrhiza fingerprint differences.To explore the overall composition of sweeting and none Salvia miltiorrhiza in vivo similarities and differences.(4)with antioxidant activity,myocardial protection,cerebral protective effects of three aspects of the comprehensive comparative analysis of efficacy of sweeting and none Salvia miltiorrhiza,efficacy from chemistry to simulate the body's cells of two different levels of sweeting Salvia miltiorrhiza were studied.Results:(1)Determination of "Danshen samples sweating" process of continuous dynamic sampling of cryptotanshinone,tanshinone I,the content of tanshinone IIA and salvianolic acid B,The results is in the process of "sweating",liposoluble constituents tanshinone(cryptotanshinone,tanshinone I and tanshinone IIA)content and water-soluble constituents of salvianolic acid B both had significant changes;the content of tanshinone and salvianolic acid B is accompanied by a "sweating" increase in the number of volatility rises,and two kinds of components in the similar upward trend;"sweating" in the process of total tanshinone content increased 80.15%,the content of salvianolic acid B increased 49.36%.(2)Using HPLC to establish the sweating fingerprints of Radix Salviae Miltiorrhizae,through chromatographic fingerprint similarity evaluation system(2004A version),showing 35 common peaks.The similarity between samples of sweeting Salvia transpiration mean was0.9672±0.0280,the overall composition is relatively stable between samples of sweeting Salvia miltiorrhiza.The HPLC fingerprint records based on the information,using principal component analysis(PCA)dimensionality reduction of multidimensional information on sweeting and none of Salvia miltiorrhiza samples.The dimension calculation result can see two groups of samples can be separated.Combined with the analysis of HPLC fingerprint analysis and principal components in sweeting and none Salvia miltiorrhiza on the basis of similarity value is high.Can distinguish the separation of two groups of samples.The results show that the overall similarity between sweeting and none of Salvia miltiorrhiza samples,but there are differences.(3)Through the chromatographic fingerprint similarity evaluation system and the common peaks were analyzed by variance analysis,After administration of sweeting and none Salvia miltiorrhiza in rat plasma fingerprint similarity average in the range of >0.800,a total of 14 only 2 peaks had significant differences(P < 0.05).Sweeting and none Salvia miltiorrhiza administration under the normal state of rats,the body composition into the overall similarity between the components of the plasma in the overall content of no significant difference.(4)With IC50 as the evaluation index of LPI lipid peroxidation,DPPH radical scavenging,superoxide radical SRS clear rate of three in the system of comprehensive in antioxidant activities of sweeting and none salvia miltiorrhiza were determined.The results were better than the antioxidant activity of non sweeting Salvia miltiorrhiza.Based on the H2O2 induced damage of H9C2 cells after oxygen glucose deprivation and reperfusion(OGD/R)injury was observed in PC12 cells in cell morphology,cell viability and cell apoptosis detection indexes,the results for the same concentration of sweeting of Salvia miltiorrhiza on the injury of H9C2 cells,PC12 cells with protection is better than that of none Salvia miltiorrhiza.Conclusion: Salvia miltiorrhiza "sweating" treatment can improve cryptotanshinone,tanshinone I,the content of the main active ingredients of tanshinone IIA and salvianolic acid B.After “sweating” treatment with Salvia miltiorrhiza,overall chemical composition did not change,and the HPLC fingerprint of no new material peak;similarity between sweeting and none salvia miltiorrhiza is high.Based on HPLC fingerprint and principal component analysis,the two kinds of Salvia miltiorrhiza samples can be separated and identified;the overall similarity between sweeting and non sweeting of Salvia miltiorrhiza samples,but there are different.Compared with the non sweating,the sweeting in the anti oxidative activity of Salvia miltiorrhiza,myocardial protection,the efficacy of cerebral protective effect,are better. |
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