Effect Of Dalbergiae Odoriferae On The Salvia Miltiorrhiza Absorption Dynamic Influence And Mechanism Research

Investigation on the compatible regulation is a difficult topic in the research area of tradition Chinese medicine(TCM). Chinese herb medicine compound prescription are constitute of two or more crude drugs, choosing the proper dose to cure the disease according to basis of theory of traditional Chinese medicine. Herb couple is a link between single herb and complex prescription, through reaearch the herb couple to proclaim the scientificalness and rationality. Jun-Shi couple herb is the special compatibility in Chinese herb prescription, it has simple constitute and easy to research. Thus, investigate the interaction and mechanism of Jun-Shi couple herb has a significant role in proclaiming the compatible regulation. Meanwhile, it also plays a very important role in enriching and developing the compatible theory of complex prescription and development of innovative medicine. In this thesis, JiangXiang was selected as the Shi herb in the prescription of"Coronary Danshen Dropping Pills", it contains volatile oil and flavone, which were used as marker to study effect CTS and SalB on the absorbtion kinetics and researching it promoted the absorbtion mechanismPart One Eestablishment and Validation of LC-MS/MS, UPLC Methods to Determine the Concentration of CTS in Caco-2 and Rat Plasma.The steady and reliable LC-MS/MS and UPLC methods for the quantitative analysis of CTS were establish in Caco-2 and rat plasma. Under this analytic situation, the outcomes chromatograms of CTS exhibited well-resolved peaks. The results shows that the calibration curves were linear in the rang of 5~640ng/mL(Caco-2), 2.5~2400ng/mL(plasma), all with r>0.99. The detecction limits of the Caco-2 and plasma are 5ng/mL, 2.5ng/mL, respectively. The extracting recovery of analytes were more than70%, intra-batch and inter-batch RSD were less than 13% in Caco-2 amd freeze-thaw stability, matrix effect and quality control were in accord with requirments of biological sample analysis of Food and Drug Administration. All the above demonstrated that the sensitive, reliable, specfic and accurate methods could be used in determination of CTS in samples.Part Two Shi Herb JangXiang Effected Absorption kinetics of CTS and SalB in Jun Herb DanShen in vivoTo study the volatile oil and flavone from JiangXiang promoted the absorbtion of CTS and SalB in vivo. Experiments were conducted the contents of CTS and SalB in DanShen extract using the HPLC method and detected the content and composition of volatile oil using the GC and GC-MS. Based on this, the pharmacokinetics differences of CTS and SalB are analyzed by the proposed method after intragastric administration of Dan Shen extract and Dan Shen extract-volatile oil group. The results show that the concentration of CTS and SalB are 1.175±0.472 mg/g, 26.089±4.718mg/g in DanShen extract. In addition, the composition and content of volatile oil from JiangXiang were detected and identified five substance, the nerolidol has the hightest content about 56.30%, 46.53%, 46.83%, respectively, in the standard substance of volatile oil, saturation volatile oil of Hank's and saturation volatile of culture medium. Through data statistics of the DanShen extract, DanShen extract-volatile oil group and DanShen extract-naringenin shows that t1/2α,t1/2β,AUC,Cmax,V1/F,CL/F have significance difference. Because of the volatile oil and naringenin existence that could increase the amount in blood and increase the bioavailability of CTS and SalB, prolong the absorbtion time and promote the CTS and SalB into the issue. These results verified the action of enhancing therapeutic effect and guidance of JingXiang, which provided the evidence to elucidate the promoted absorbtion of JiangXiang.Part three The Volatile oil and Flavone of JiangXiang Effected the CTS and SalB involved the Absorbtion in Caco-2 and MDCK ModelThe purpose of this part to estabilish the Caco-2 and MDCK model, and to further illuminate the uptake characteristic of CTS and SalB in the model. Experiments were conducted to examine time-concentration and temperature-dependent elements of CTS and SalB uptake, and the effect of volatile oil and flavone to CTS and SalB uptake in the Caco-2 and MDCK model. In the present or absence of CyclosporinA(CsA)was examined to evaluate whether P-gp contributed to the CTS and SalB efflux transport across Caco-2, thereby to confirm the absorbtion mechanism of CTS and SalB. Because of volatile oil and naringenin apply to intragastric administration, the content of CTS and SalB uptake have a significant increase and the peak ahead of the time. Western blot show that volatile oil has no effect about the P-gp protein express, but the naringenin has decreased the P-gp protein express.These results indicated the uptake of CTS and SalB by Caco-2 and MDCK were found to be a time-concentration and temperature-concentration. The mechanism absorbtion of CTS in small intestine is based on the active transport and passive diffusion participate in, The mechanism absorbtion of SalB is based on the passive diffusion. The volatile oil from JingXiang nonspecific promote the CTS and SalB absorbtion and naringenin specificity promote the CTS and SalB absorbtion in the small intestine.