| Chapter one, Literature Study.A synthetical review was given to the present research of Shaofu-Zhuyu Decoction(SFZYD) application and thinking for metabonomics in some scientific problem of traditional chinese medicine as well as the research progress of transdermal delivery systems in China.Chapter two, Process Research of Shaofu-Zhuyu Transdermal Drug Delivery System.In Section 1, Improved Franz diffusion cells with isolated rat abdomen skins were carried out to experiment on the transdermal delivery of nine active components, including ferulic acid, paeoniflorin, albiflorin, protopine, tetrahydropalmatine and moreover, a method based on principal component analysis for the study of transdermal delivery of multiple components is established, which is confirmed to be suitable and objective for the study of transdermal delivery of Chinese Medicine. The result shows that 2.5% azone with 2.5% propylene glycol manifested the best effect.In Section 2, two different matrixes, including lipophilic matrix and hydrophilic matrix are compared using the transdermal delivery experiment. The result indicates that the major components in SFZYD show obvious difference on transdermal delivery in these two matrixes. In lipophilic matrix, the realease of components is fast followed by slow, which accords with the Higuchi Equation, when it applied to be uniform and even in hydrophilic matrix. The difference of accumulative penetration amount of typhaneoside, protopine and tetrahydropalmatine in two matrixes is similar, whereas others show significant difference. So as the consequence, hydrophilic matrix is considered as a better matrix.In Section 3, a synthetical optimization of the patch, including excipient, procedure, drug loading, is carried using above established methods and experiments. As a result, the matrix consists of CMC-Na 10g, PVA 15g, PVP K-30 25g and glycerol 10g; 10 fold water,70℃ with 200 rpm for stir is suitable for the procedure; The maximum loading drug is as high as 4% for SFZYD, and furthermore,7.5% penetration enhancers manifested the best effect.Chapter three, Pharmacodynamics study of Shaofu-Zhuyu Transdermal Drug Delivery System.hot plate test is used to validate the pharmacodynamics efficacy in terms of the pain thresholds. The results show that high dose group (4% SFZYD) increases the pain thresholds of mice significantly, while abirritation effect of the medical and low group is unconspicuous.Chapter four, metabolism and its mechanism study of Shaofu-Zhuyu DecoctionIn Section 1, a sensitive and rapid UPLC-MS/MS method was developed for simultaneous quantification of paeoniflorin, albiflorin, ferulic acid, tetrahydropalmatine, protopine, typhaneoside and senkyunolide I in Beagle dog plasma after oral administration of the Shao-Fu-Zhu-Yu Decoction. After validation, this method was successfully applied to a pharmacokinetic study. The results showed that the apparent plasma clearance of paeoniflorin, albiflorin, typhaneoside and senkyunolide I were significantly higher than others. Double peak was observed in plasma concentration curves of tetrahydropalmatine, the ferulic acid had a good absorption in Beagle dog plasma, and senkyunolide I was detected in plasma from the first blood sampling time (15 min) and rapidly reached Tmax. The compound of typhaneoside has a low bioavailability according to the results.In Section 2, establish the model of primary dysmenorrhea of cold-stagnation and blood-stasis type in ice bath and continuous injection of epinephrine; taking estrogen and oxytocin in thepreparation of primary dysmenorrhea model rats. Using UPLC-QTOF/MS series technique combined with multivariate statistical modeldiscrimination methods, founding normal rats and model rats were divided into two classes, and presented a cluster state, states that primary dysmenorrhea of cold-stagnation and blood-stasis type rat model successfully reproduced. The biochemical indexes related to primary dysmenorrhea:prostaglandins, oxytocin, neuroendocrine hormone β-endorphin, progesterone, estradiol and endothelin changed before and after the model making.In Section 3, In the plasma, ten endogenous metabolites, contributing to the separation between the groups, were identified by comparing with authentic standards or based on their molecular ion information as well as the fragments of corresponding product ion. In the urine, the detected significant variables in the positive ion mode. Fifteen endogenous metabolites were tentatively identified by the methods described above. Furthermore, eight endogenous metabolites in plasma and thirteen endogenous metabolites in urine were significantly affected by SFZYD. The metabolic pathway analysis with MetPA revealed that the identified biomarkers in plasma and urine were critical for the host responding to primary dysmenorrhea of cold stagnation and blood-stasis type, which was responsible for pentose and glucuronate interconversions, glycerophospholipid metabolism and steroid hormone biosynthesis.The levels of indicators were changed significantly after the treatment of SFZYD. For hormone indicators, SFZYD reduced significantly the levels of Estradiol, and SFZYD obviously increased the levels of Progesterone. The elevated inflammatory factors levels of PGF2 and Endothelin were inhibited significantly after the treatment of SFZYD. The elevated neuro-transmitter Oxytocin was reduced significantly. These data implied that SFZYD regulated multiple biochemical indicators related to neuro-endocrine-immune (N-E-I) system and exhibited an integral efficacy.In Section 4, paeoniflorin, a major bioactive compound in SFZYD, regulates the MAPK signaling pathway by inhibiting the expression of IL-1β, IL-2, IL-10, IL-12, TNFa, INFy, C-jun and C-fos. |
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