| Rhododendron dauricum L.is the dry leaf of Rhododendrons of the family Azaleae.It is a green shrub growing for many years.It has unique fragrant smell,bitter taste,slight spicy,and belongs to the lung and spleen meridian.At present,the preparation with Rhododendron dauricum L.as raw material is only used for the treatment of respiratory diseases,and the treatment range is narrow,which affects its development and utilization.It is urgent to make breakthroughs in biological activities,while antioxidant is related to a variety of biological activities.Drosophila has the advantages of short growth cycle,easy observation of physiological characteristics and low feeding cost,making it an ideal animal model for antioxidant experiments.Therefore,this study introduced drosophila model to study the antioxidant effect of Rhododendron dauricum L.for the first time,and established the spectral effect relationship model of Rhododendron dauricum L.It provides experimental research basis for expanding the biological activity of Rhododendron dauricum L.and further developing its medicinal resources.The specific research contents are as follows:In this experiment,the extract of Rhododendron dauricum L.was separated and purified to provide reference for the later production of fingerprint.The antioxidant activity of Rhododendron dauricum L.was studied using drosophila as experimental animal.Of 10 batches using HPLC method with red mountain medicinal materials were analyzed,and the chromatographic conditions were optimized and complete methodology,establish.HPLC fingerprint of Rhododendron dauricum L,use the software named fingerprint similarity calculation of Traditional Chinese Medicine to calculate the similarity,and use cluster analysis and principal component analysis method of fingerprint pattern recognition research;Using DPPH free radical scavenging assay to To detect antioxidant capacity of Rhododendron dauricum L,Rhododendron dauricum L.based on grey correlation analysis method the chemical composition of the fingerprint characteristic peak represents information relationship with the antioxidant effect,finally according to the correlation between the size to determine the chemical composition of the fingerprint characteristic peak represents against oxidation degree of contribution to the therapeutic effects.Five flavonoids were isolated,which were Farrerol（1）,3’-hydroxylfarrerol’（2）,8norrhododene（3）,Quercetin（4）,Hyperin（5）.Among them,compounds 1,4 and 5 almost represent the flavonoids found so far in this herb,which can be used as reference substances for fingerprint peak identification after repeated purification.The results of antioxidant test in drosophila showed that compared with the control group,half of the death time,average life and maximum life of drosophila in the treatment group were prolonged（P<0.05）.The climbing ability of drosophila in the treatment group was increased（P<0.05）.The activities of T-AOC and SOD were increased（P<0.05）,while the content of MDA was decreased（P<0.05）,indicating that Rhododendron dauricum L.had better antioxidant capacity.Using farrerol as the reference peak,16 common peaks were determined,and the similarity results were 0.913,0.957,0.949,0.947,0.958,0.971,0.931,0.956,0.977,0.933,which met the relevant requirements.The results of cluster analysis showed that the medicinal materials of Rhododendron dauricum L.were divided into four categories,and the medicinal materials with the same harvesting time were classified into one category,suggesting that the quality of Rhododendron dauricum L.might be correlated with the harvesting time.Principal component analysis of the common peak is divided into four main component,and a greater influence on the quality of the first principal component of full mountain red,according to the first principal component of the load quantity size 3,7,8,9（quercetin）and 10（hyperin）,11,12,13,15 and 16（farrerol）peak on behalf of the chemical composition of red play an important role in the quality control in full mountain;The results of the score map of Rhododendron dauricum L.herbs showed that 10 batches of herbs could be divided into 4 categories,and the results were consistent with cluster analysis.Grey correlation degree showed that 16 common peak and the correlation between the antioxidant efficacy have 12 above 0.6,respectively,peak 2,3,5,6,8,9（quercetin）,10（hyperin）,11,12,14,15,16（farrerol）are greater than 0.7 there are seven respectively for peak 3,6,8,9（quercetin）and 10（hyperin）,12,16（farrerol）,The contribution of antioxidation effect is large.According to the UV spectrum analysis,the structure of the associated peaks has not been determined.Peak 11,14 and 15 may be the compounds what are not dihydroflavones,peaks 2,5,6,8 and 12 may be flavonols,and peaks 3 may be the compounds what are not dihydroflavones and flavonols.Conclusion:antioxidant experiment of Drosophila melanogaster shows that Rhododendron dauricum L.has better antioxidant effect.The established HPLC fingerprints of 10 batches of Rhododendron dauricum L.were imported into the similarity evaluation system of Traditional Chinese medicine fingerprint,and the similarity results met relevant requirements.Cluster analysis and principal component analysis showed similar results,indicating that the established fingerprints can be used for quality control of Rhododendron dauricum L.and the follow-up study of spectral effect relationship.The results of grey correlation analysis showed that the antioxidant activity of Rhododendron dauricum L.was the result of "effective substance group",and quercetin,hyperin and farrerol were identified as the components that played antioxidant effect. |
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