Study On Anti-Inflammatory Specturm Effect And Quality Evaluation Of Bupleuri Radix

Objective:The Chinese medicine Bupleuri radix has a significant effect on the evacuation and antipyretic effect.It is often used to solve the problem of heat dissipation,and the body fever is closely related to inflammation.Modern pharmacological studies have confirmed that Bupleuri radix has obvious anti-inflammatory effects,but the complex source,origin and processing methods of Bupleuri radix can cause quality differences,which in turn affects the anti-inflammatory activity of Bupleuri radix and its compound.Therefore,it is necessary to study the anti-inflammatory spectrum effect relationship of Bupleuri radix,screen the anti-inflammatory effect of Bupleuri radix medicinal substances,and further establish the quality evaluation method of this kind of ingredients to control the quality of Bupleuri radix.Methods:1.With 20 batches of Bupleuri radix from different habitats as research objects,the quality evaluation method of HPLC-DAD dual-wavelength fingerprint was established,and the fingerprint information of Bupleuri radix from each place was obtained.The components of common peak were determined by HPLC-Q-TOF-MS.2.After mouse RAW264.7 cell inflammatory model was induced by lipopolysaccharide（LPS）,NO,IL-1β,IL-6 and TNF-αinflammatory factors related to anti-inflammatory activity of Bupleuri radix were assessed in vitro.Multivariate linear regression and gray correlation analysis were used to construct the"spectrum-effect relationship"of Bupleuri radix anti-inflammation profile,and to select the characteristic peaks related to the anti-inflammatory activity of Bupleuri radix in vitro.HPLC method for determination of anti-inflammatory active components of SSa,SSe,SSd,SSb₂ and SSb₁ in Bupleuri radix samples was studied.The method was applied to determine the content of five anti-inflammatory active components in 20batches of Bupleuri radix samples from different producing areas.Result:1.The similarity software analysis was used to determine 13 common peaks.The similarity between Bupleuri radix peaks from different producing areas was found to be between 0.108 and 0.98,indicating that the nature of Bupleuri radix samples was significantly different,which may attribute to the growth of different base sources,different producing areas and different management methods.Principal component analysis showed that there were 4 principal components,and the cumulative contribution of the three variances was 84.707%,of which the three components were SSa,SSd and F11.The 13 common peaks were assigned by HPLC-Q-TOF-MS,and 12chemical components were identified,including 9 triterpenoid saponins,1 flavonoid and 2 Volatile oil component.Wherein F1 is Isoquercitrin,F2 is SSc,F3 is SSf,F4 is SSa,F5 is SSe,F6 is 3’-acetyl SSa,F7 is SSd,F8 is 3’-acetyl SSd,F10 is Methyl linoleate,F11 is Linoleic acid,F12 is SSb₂,F13 is SSb₁.The results showed that the method can identify chemical constituents of common peaks in Bupleuri radix.2.With LPS to stimulate RAW264.7 to establish an in vitro cell inflammatory model,inflammatory factors NO,IL-1β,IL-6 and TNF-αwere chosen as indicators to study in vitro anti-inflammatory activity of 20 batches of Bupleuri radix from different producing areas.Furthermore,multiple linear regression analysis and gray correlation analysis were used to analyze the peak area data of the common peaks in 20 batches of Bupleuri radix fingerprints and in vitro anti-inflammatory activity of Bupleuri radix.The results showed that there were significant differences in anti-inflammatory activity of Bupleuri radix samples from different habitats.Among them,Bupleuri radix in Hailaer,Qinghai and Chengcheng County had strong anti-inflammatory activity in vitro;Sichuan,Pingliang County,Kangle County of Gansu Province has poor anti-inflammatory activity in vitro.The resulted components which were screened for its anti-inflammatory activity were SSa,SSe,3’-acetyl-SSa,SSd,SSb₂,SSb₁ and F9.3.A HPLC dual-wavelength determination method for five anti-inflammatory active ingredients in Bupleuri radix was established.SSa,SSe,SSd,SSb₂ and SSb₁ were selected as detection indexes.The results showed significant differences between different producing areas,cultivated products and wild products.If total amount of five saponins was used to evaluate the anti-inflammatory activity of Bupleuri radix,the cultivated products from Qinghai were better,followed by Chengcheng in Shaanxi and Ganle.To provide an experimental basis for the determination of anti-inflammatory components in vitro,this method controls and evaluates the quality of Bupleuri radix based on anti-inflammatory activity.Conclusion:1.Through the in vitro anti-inflammatory spectrum effect study of Bupleuri radix,the active ingredients of Bupleuri radix in vitro anti-inflammatory effect were screened out.2.Based on the selected active ingredients,HPLC method for determination of five components of Bupleuri radix was established to provide a basis for quality control and evaluation of Bupleuri radix anti-inflammatory active ingredients.