| Objective: Combined with the study of the "spectrum" and "effect" to establish the spectrum-effect relationdship.Clarify the material basis of green Forsythiae Fructus anti-inflammatory and old Forsythiae Fructus hepatoprotective effect based on the spectral-effect analysis method.And explore the new ideas of the research on the quality control based on the difference between the two active substances.Method: 1.HPLC fingerprints of anti-inflammatory and hepatoprotective active parts of Forsythiae Fructus were established by high performance liquid chromatography(HPLC),and the peaks and its area information of the common peaks was obtained.2.The spectral correlation of anti-inflammatory activity of different polar parts of green Forsythiae Fructus was studied by the method of gray correlation and bivariate correlation analysis.The chromatographic peaks with great contribution to the efficacy were found out,and then clarify its information on composition preliminarily.3.The inhibitory effect of green Forsythiae Fructus and old Forsythiae Fructus on serum ALT and AST in CCl4 liver injury rats and its effect on histopathology were compared.Thereafter conduct the efficacy experiments aim at multi-batchs of hepatoprotective activity part.4.The spectral correlation of the hepatoprotective active parts of green Forsythiae Fructus and old Forsythiae Fructus was studied by the partial least squares regression analysis method,and the chromatographic peaks with large contribution to efficacy were obtained,the components were identified by reference substance information.Results: 1.The results of HPLC fingerprints of anti-inflammatory active parts of green Forsythia suspensa: There were 31 common peaks in the polar parts of green Forsythia suspensa,and there were 27 common peaks in the part of the water extract,26 common peaks in the Ethyl acetate site,19 common peaks in the N-butyl alcohol site and 12 common peaks in the water site.The results of HPLC fingerprints of hepatoprotective active parts of Forsythia suspensa: The Q-1 part have 21 common peaks,Q-1 part have 14 common peaks,Q-3 part have 14 common peaks,Q-3 part have 19 common peaks,Q-3 part have 17 common peaks,L-1 has 14 common peaks,L-2 has 9 common peaks,L-3 has 5 common peaks,and L-4 has 7 common peaks.2.The correlation analysis result of anti-inflammatory effects of different polar parts: Gray correlation analysis showed that 7 peaks with large contribution to pharmacodynamics,the different peak ranked as No.23,25,31,24,26,30.Bivariate correlation analysis showed that there was a significant negative correlation between the No.23 peak content and TNF-α(P<0.01),between the No.18 and No.13 peaks content with IL-6(P<0.01,P<0.05),between No.22 peak content with IL-1β(P<0.05),between the No.7,No.13 and No.6 peaks content with IL-1β(P< 0.05).That is to say,the higher contents of chemical components of these peaks,the lower of TNF-α,IL-6 and IL-1β,and the better was the anti-inflammatory effect.According to the results of two kinds of analysis,it was concluded that the peaks of 23,13,25,31,etc.had the greatest contribution to the anti-inflammatory effect.3.The results of Hepatoprotective active site: Compared with the model group,the water extracts of the green Forsythiae Fructus and the old Forsythiae Fructus,and the ethyl acetate extracts could significantly decrease the ALT activity(P <0.05),the activity of AST was significantly decreased(P <0.01,P <0.05)by the water extracts of the green Forsythiae Fructus and the old Forsythiae Fructus,and the N-butyl alcohol extract.Histopathological observation of liver tissue showed that from the Q-1 group to Q-4 group,L-1 to L-4 group had different degree of mitosis compared with the model group.4.The results of hepatoprotective effects of the green Forsythiae Fructus and the old Forsythiae Fructus water extract based on partial least squares regression analysis: According to the VIP chart of ALT,the peak of the contribution to the drug efficacy is: No.11,3,13,9,6,18,19,5,21.From the chart of standard regression coefficient,most of the peak are negatively correlated with ALT,which is associated with the hepatoprotective function positively,with the increase of their content,the ability of anti liver injury may be enhanced.According to the VIP diagram of AST,the peak of the contribution to the efficacy of the order is: No.11,6,3,9,19,21,5,2,13,16,18.It can be seen from the standardized regression coefficient,No.3,21,19,5,6,2,18,13 peaks were negatively correlated with AST,which is associated with the hepatoprotective effect,with the increase of their content,the ability of anti liver injury will be enhanced.From the above results we can see that the maximum correlation(degree of association and correlation coefficient)peak with the anti-inflammatory effect of green Forsythiae Fructus: No.23(forsythin),13(unknown),25(quercetin),31(phillygenin)and etc.The maximum correlation peak with the hepatoprotective effect of old Forsythiae Fructus may be: No.3(unknown),21(phillygenin),19(quercetin),5(unknown),6(unknown),18(forsythin),13(pinoresinol-β-D-glucoside)and etc.Conclusion: The ethyl acetate part was the anti-inflammatory activity part of green Forsythiae Fructus,and the main activity material basis may be forsythin,quercetin and phillygenin and other medium polar components.The water extract was the hepatoprotective activity part of green Forsythiae Fructus and old Forsythiae Fructus.and the main activity material basis may including the phillygenin,quercetin,forsythin and pinoresinol-β-D-glucoside and other polar components,suggesting that the hepatoprotective activity was the commom effect of a variety of different polar components of Forsythia Fructus.It can be seen the material basis of Forsythia Fructus anti-inflammatory and hepatoprotective function is different. |
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