| Objective:To investigate the antidepressant effects and the optimum preparing technology of Chinese medicine formual suanzaorenhehuan(CSH)was chosen. To searched the best compatibility by uniform design method and establish a quality control method of Chinese medicine formual suanzaorenhehuan. To screen the antidepressant effective fraction of the Chinese medicine formual suanzaorenhehuan by pharmacodynamics.Method:1.The forced swimming test(FST), tail suspension test(TST), open field test(OFT) and Reserpine-induced hypothermia test were carried out to determine the antidepressant effects in mice. Mice immobility duration in FST and TST, spontaneous activity frequency in OFT, and body temperature decrease in Reserpine antagonistic test were observed. To investigated the antidepressant effects and screen the optimum preparing technology of CSH. The preparing technology include water extracted、alcohol extracted、alcohol extracted with X-5or AB-8macroporous resin.2. The uniform design method combined with pharmacodynamics was used to evaluate the antidepressant effect of the formula and to screen the optimnm dose of the four herbs in CSH by using multivariate statistics.3.Jujuboside A、B and peoniflorin was identified by TLC. The total flavonoids content in the formula was determined by UV spectrophtomotry. A method for determining Jujuboside A in CSH had been established by using HPLC-ELSD.4. According to the literature, CSH principally contain saponin and flavonoids. The forced swimming test, tail suspension test, open field test and Reserpine-induced hypothermia test were carried out to compare the antidepressant effects in mice between total saponin and total flavonoids.Result:1.The group of AB-8could significantly shorten mice immobility time in FST and TST. There are no significant differences on mice spontaneous activity during each group. Compared with the model group, the group of AB-8could remarkably inhibit the decrease of body temperature in Reserpine antagonistic test at1th and4th hour.2.The optimal doses of the four herbs in CSH were screened out theoretically by the uniform design approach combining pharmacyodynamics: Suanzaoren24.00g:Albizia14.00g:TGP6.00g:Bo Ziren10.00g.3. The samples showed the similar spot with reference substance and medical material on TLC. The content of total flavonoids in the formula was determined by UV spectrophtomotry, the concentration was linear relationship with absorbance in the range of8.5～25.5μg·mL-1. The equation of linear regressin was Y=0.0225X+0.0229. The average recovery was101.4%, and RSD was0.61%. The content of Jujuboside A in the formula was determined by HPLC-ELSD, the concentration was linear relationship with absorbance in the range of0.6-3.0μg. The equation of linear regressin was Y=1.2299X+13.295. The average recovery was99.98%, and RSD was1.19%. The results show that this method is accurate, sensitive and reproducible. And it proved to be feasible for quality control of total flavonoids.4.The groups of total saponin and formula could significantly shorten mice immobility time in FST and TST. There are no significant differences on mice spontaneous activety during each group. The group of total flavonoids could also shorten mice immobility time in FST and TST, but there is no significant difference. Compared with the model group, the groups of total saponin, formula could remarkably inhibit the decrease of body temperature in Reserpine antagonistic test at4th hour.Conclusion:The extract method combined pharmacodynamic index has the merit of being related to the function and symptom directly. The the optimum technology of preparing CSH is suitable for industry production. The optimum dosage of CSH is verified by pharmacodynamics test. The verifying result show the optimized prescription was significantly effective. It would be possible to use uniform design combined with pharmacodynamics method in defining the optimal dosage of four herbs in CSH. The determination of total flavonoids and Jujuboside A was simple, accurate and could be used to control the quality of total flavonoids and Jujuboside A in CSH. The pharmacodynamic result show total saponin is the active fraction. |
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