Résumé
AIM To investigate the effects of drying temperature,growing area and plucking time on hederacoside C,α-hederin from leaves of Hedera helix L..METHODS Three batches of H.helix leaves plucked in different time from two growing areas were dried in a vacuum oven to the constant weight at 60 ℃,70 ℃,80 ℃,90 ℃ and 105 ℃,respectively.Two saponins in the processed leaves were determined by HPLC.The powders of the processed H.helix leaves of different batches were mixed with proper ratios,which were determined by least squares optimization method with constraints.RESULTS The content of hederacoside C in the processed H.helix leaves of the three batches increased while that of α-hederin decreased with increasing temperature.The relative error between measured value and desired contents of hederacoside C and α-hederin in the mixed H.helix leaves was less than 5.5%.CONCLUSION The effects of three factors on the content of two saponins in the H.helix leaves are in the order of drying temperature,growing area and plucking time.Mixing processed H.helix leaves of different quality statues reasonably can control the contents of two saponins in a certain range.
Résumé
Objective: According to the results of preliminary cellular pharmacological experiments, 13 active fractions, including Yaotongning Capsule (YTNC) whole formula, were designed to evaluate their in vitro and in vivo anti-inflammatory and analgesic activities to look for candidate prescriptions for rheumatism treatment. Methods: The in vitro anti-inflammatory and analgesic effects of the prescriptions were estimated by detecting their ability to inhibit 3H-bradykinin binding to its receptor and ability to inhibit 3H-naloxone binding to opioid receptors, respectively. And the tests were conducted by a liquid scintillation counter to check the content of the isotope 3H as evaluating indicator. Ear edema model in mice induced by dimethylbenzene test, hot plate test, and acetic acid writhing test in mice were conducted to evaluate the in vivo anti-inflammatory and analgesic activities of the prescriptions. Results: YTNC whole formula had good in vitro and in vivo anti-inflammatory and analgesic effects. However, the prescription containing equal amount of Ephedra sinica alkaloids, Glycyrrhiza uralensis aponins, and Carthamus tinctorius flavones (Sample VI) as well as the optimized and decomposed formula of YTNC (Sample VIII), which was designed by reducing 90% of Strychnos nux-vomica alkaloids from YTNC's decomposed formula composed by alkaloids, flavonoids, and saponins in YTNC, performed more excellent in vitro anti-inflammatory and analgesic effects than whole formula of YTNC. However, Sample VI did not exhibit obvious in vivo analgesic effect, but the in vivo anti-inflammatory and analgesic activities of the optimized and decomposed formula of YTNC (Sample VIII) were more significantly better than those of YTNC whole formula. It also had significantly better analgesic effect than aspirin (the positive control drug) in acetic acid writhing test. Conclusion: The optimized and decomposed formula of YTNC could be a candidate drug for rheumatism treatment. The anti-inflammatory and analgesic effects, safety, and quality control ability of YTNC could be improved by reducing the varieties of material medicines and active fractions and the content of S. nux-vomica alkaloids in YTNC.
Résumé
Because of the numerous varieties of herbal species and active ingredients in the traditional Chinese medicine(TCM),the traditional methods employed could hardly satisfy the current determination requirements of TCM.The present work proposed an idea to realize rapid determination of the quality of TCM based on near infrared(NIR)spectroscopy and internet sharing mode. Low cost and portable multi-source composite spectrometer was invented by our group for in-site fast measurement of spectra of TCM samples. The database could be set up by sharing spectra and quality detection data of TCM samples among TCM enterprises based on the internet platform.A novel method called as keeping same relationship between X and Y space based on K nearest neighbors(KNN-KSR for short)was applied to predict the contents of effective compounds of the samples. In addition,a comparative study between KNN-KSR and partial least squares(PLS)was conducted. Two datasets were applied to validate above idea:one was about 58 Ginkgo Folium samples samples measured with four near-infrared spectroscopy instruments and two multi-source composite spectrometers,another one was about 80 corn samples available online measured with three NIR instruments. The results show that the KNN-KSR method could obtain more reliable outcomes without correcting spectrum.However transforming the PLS models to other instruments could hardly acquire better predictive results until spectral calibration is performed. Meanwhile,the similar analysis results of total flavonoids and total lactones of Ginkgo Folium samples are achieved on the multi-source composite spectrometers and near-infrared spectroscopy instruments,and the prediction results of KNN-KSR are better than PLS. The idea proposed in present study is in urgent need of more samples spectra, and then to be verified by more case studies.
Résumé
Objective: To evaluate the in vitro effects of different combinations of the active fractions in Yaotongning Capsule (YTNC) on anti-inflammation, immunity, and activating blood circulation, and the interactions among these active fractions. Methods: Six samples were prepared by adding flavonoids, saponins, volatile oils/aqueous extracts, and polysaccharides, which were extracted from the material medicines in YTNC, to the alkaloids of Strychnos nux-vomica (Ma Qian Zi) and Ephedra sinica (Ma Huang) in turn according to the formulating principle of YTNC, combination chemistry concept, and their importance in the formula. The effects of the six samples on the production of lipopolysaccharide (LPS)-induced prostaglandin E2 (PGE2) in Ana-1 cells, concanavalin (ConA)- induced interleukin-2 (IL-2) in splenocyte, and nitrogen monoxide (NO) in human umbilical vein endothelial cells (HUVEC) were explored by detecting their half-maximal inhibitory concentration (IC50) or half-maximal effective concentration (EC50). An innovative method based on least square optimization was applied to calculate the additive IC50/EC50 value of each sample. The interactions of the active fractions in these samples were investigated by comparing the additive IC50/EC50 values with the corresponding experimental IC50/EC50 values. Results: All the six samples could inhibit the production of PGE2 and promote the secretion of IL-2 in the investigated concentration range. Only sample III consisting of the alkaloids, flavonoids, and saponins from YTNC had the EC50 value of promoting NO production. Moreover, the sample also displayed excellent activities in the three models of LPS-induced PGE2 production, ConA-induced IL-2 production, and NO production, which could reflect the capability of anti-inflammation, enhancing the body's immunity, and activating blood circulation, respectively. Different combination of the active fractions from YTNC produced the different interaction types in the three models. And the active fractions in sample III, i.e., the alkaloids, flavonoids, and saponins from YTNC, generated high synergistic effects in all the three models. Conclusion: All the six samples derived from the different combination of the active fractions from YTNC show the good activity of inhibiting PGE2 production and promoting IL-2 secretion. Therefore, the YTNC formula is reasonable and has good activity of anti-inflammation and immunity. However, the YTNC formula still could be improved by reducing the categories of its active fractions because the combination of the alkaloids, flavonoids, and saponins exerted better activities than the other samples and the three kinds of active fractions produced an excellent synergistic effect in all the three models. Synthetically evaluating various activities of YTNC in the present work may lead to develop new Chinese materia medica for rheumatic arthralgia treatment, which is more effective and easily controllable in quality.