Anti-prostatic Hyperplasia And HPLC Spectral Relationship Of Saxifrage

Objective:This work investigated the high performance liquid chromatography(HPLC)fingerprints of aqueous extracts fiom Saxifraga stolonifera(AESS),which covered 24 different regions.On this basis,we were screening differences samples for study its pharmacodynamics in anti-benign prostatic hyperplasia activities.The spectrum-effect relationships between HPLC fingerprints and anti-benign prostatic hyperplasia activities were investigated using Gray Correlation Analysis(GRA)and partial least squares regression(PLSR)to research the main active components,which makes preparation for investigating the mechanism of the S.stolonifera extracts in the future.Methods:1?Establishing fingerprint of AESS by HPLC:The results of our previous research "HPLC fingerprint for aqueous extraction of Saxifraga stolonifera Meerb"was used to determine the HPLC fingerprints of AESS fiom various sources.Similarity analysis(SA),hierarchical clustering analysis(HCA)and principal component analysis(PCA)were used to evaluated the common chromatographic peaks area.Meanwhile,we identified a part of chromatographic peaks.2?Screening differences samples:On the basis of 18 common peak area in chromatographic fingerprints from the 24 batches of S.stolonifera and the chemometrics including similarity evaluation,PC A,and HCA,9 batches of S.stolonifera with different chemical profiles were selected for researches on their activities and for profile-efficiency study.3?Study of different origins S.stolonifera samples in anti-benign prostatic hypeiplasia:Testosterone-induced mouse model of benign prostatic hypeiplasia(BPH)was used on anti-benign prostatic hyperplasia activities of these nine S.stolonifera samples.The model was evaluated by analyzing prostatic index(PI),serum acid phosphatase(ACP)activity,concentrations of serum dihydrotestosterone(DHT),prostatic acid phosphatase(PACP)and type ? 5-alpha-reductase(SRD5A2).4?Profile-efficacy relationship analysis:According to the results of S.stolonifera fingerprints and pharmacodynamics activities,partial least squares regression(PLSR)and Gray Correlation Analysis(GRA)were used to find the primary active components in fingerprint representing the anti-benign prostatic hyperplasia activities.Results:1?The results showed that the reference chromatogram was generated from the fingerprints of the 24 batches of S.stolonifera samples and 18 common fingerprint peaks were found contains Chlorogenic acid,Gallic acid,Protocatechuic acid,bergenin,Quercetin 5-O-?-D-glucopyra noside and Quercetin 3-O-?-L-rhamnopyranoside.The similarities between the entire chromatographic profiles of 24 batches of S.stolonifera and correlation coefficients of their chemical fingerprints were evaluated by similarity analysis.The results showed that correlation coefficients of those samples were from 0.270(Qingchang town,Guizhou)to 0.990(Qitan town,Guizhou),and the correlation coefficients between the chromatograms of samples from the same source there were big different.Results of HCA showed that 24 tested samples of S.stolonifera were divided into two main clusters.Sample nos.15 was in cluster ? and the other samples were in cluster ?,which was divided into two subgroups again.Sample nos.3 and 11 were in subgroup A and others were in subgroup B.The result of the PC A showed that first six principal components(A6,A9,A10,A13,A17 and A18)contained 82.49%of the information of the original eighteen indexes.2?As a result,we choose the nine samples of different origins as follows:A from Baiyun town,Anshun city;B from Qingchang town,Bijie city,etc.to study the anti-benign prostatic hyperplasia activities of S.stolonifera samples.3?The results show that BHP model control group had significantly difference(p<0.01)compared with the control group and positive control group all of the five pharmacodynamics indexes and histopathological examination also showed that oral administration of S.stolonifera extracts attenuated TP-induced prostatic hyperplasia.The 9 samples of S.stolonifera all show anti-benign prostatic hyperplasia activities according the results of the five pharmacodynamics indexes.4?As the results of Grey relational analysis showed the average GRG between the 5 pharmacodynamics indexes and the 18 values of peak areas and were as follow:A18>A14>A17>A16>A4>A13>A9>A15>A1>A10>A7>A5>A11>A3=A12>A6>A8>A2.PLSR results were showed in these 18 common peak areas,A14?A17 and A18 were performances negative correlation with these five indicatorsAccording to the result of GRA and PLSR,the strength of anti-benign prostatic hyperplasia bioactivity AESS was mainly affected by A14(Chloro genic acid),A17(Quercetin 5-O-?-D-glucopyranoside)and A18(Quercetin 3-O-?-L-rhamnopyranoside).Conclusion:The result indicates that chlorogenic acid,quercetin 5-O-?-D-glucopyranoside and quercetin 3-O-?-L-rhamnopyranoside might be the main active components affecting in anti-benign prostatic hyperplasia bioactivity of S.stolonifera that could provide a tool to evaluate the differences of internal quality and anti-prostate hyperplasia activities of S.stolonifera,providing a sound experimental foundation and model for the study of it.