Study On Active Essences Screening Of Anti-Helicobacter-pylor Of Zuojinwan And Modern Preparation

At present, the study of active essence of Chinese Materia Medica(CMM) is the concerned point. The theory of Traditional Chinese Medicine(TCM) revealed that the preparation and prescription compatibility can change active essence of CMM at different degree and then change the property of CMM. In order to explore the new breakthrough of study of active essence of CMM, in this paper, we investigated systematically the anti-Helicobacter pylor(Hp) active essence of Zuojingwan from three aspects of prescription compatibility, preparation and active fraction and carried out the preparing technic study of Zuojing Sustained Floating Tablets(ZJSFT), taking preparations of Coptidis Rhizoma (CR) and Zuojing similar formula as objects, adopting biothermodynamic as main method, and combining with chemical basis analysis and pharmacology experiment. Based on this, we tried to establish a new mode and method of active essence screening of heat cleaning CMM . The main results as following:1. Investigation on biothermodynamic method: The result showed that the biothermodynamic spectrogram of bacterial metabolism was well coincided with phases of biologic cultivation such as lag phase, exponential growth phase, stable phase and decline phase. The biothermodynamic spectrogram can integritly describe the bacterial metablic phases. The profile of exponential growth phase fit with linear eqution: InPt=InP0+kt. It indicated that microcalorimetry had such superiorities as actual time, on-line, repeat well, easy to manipulate.2. The study of biothermodynamic: Adopting the method of biothermodynamic, taking Escherichia coli, Shigella dysenteroae, Staphylococcus aureus, Helicobacter pylori(Hp) as pattern organism, the biothermodynamic spectrograms and parameters of aqueous extracts of the preparations of CR(Shenghuanglian, Jiangzhihuanglian, Wuyuhuanglian, Jiuzhihuanglian, Cuzhihuanglian, Yanhuanglian and Danzhihuanglian) and Zuojing similar formula(Fanzuojinwan, Zhuyuwan, Ganlusan and Zuojinwan) were determined. The result showed the biothermodynamic parameters such as growth rate constant k and heat production Q can exosyndrome the differences of biologic activity of preparations of CR and Zuojing similar formula. The sequence of k and Q : Wuyuhuanglian > Jiuzhihuanglian > Jiangzhihuanglian > Shenghuanglian > Danzhihuanglian > Yanhuanglian > Cuzhihuanglian, Fanzuojinwan> Zhuyuwan > Ganlusan > Zuojinwan. The rule of change coincided with the property of CMM, it also indicated a better method for evaluating the property or activity of CMM was put forward.3. The study of routine pharmacological experiment:Through routine vitro bacteriostasis experiment, the anti-Hp action of preparations of CR and Zuojing similar formula was studied.The result showed that there were differences between different preparations of CR and Zuojing similar formula, it also indicated that Cuzhihuanglian and Zuojingwan had the best anti-Hp activity. It verified and supported the result of biothermodynamic experiment and showed that biothermodynamic method can be served as an available method to determine the property and activity of CMM. 4. The study of chemical basis: In this paper,chemical HPLC fingerprint chromatograph of aqueous extracts of preparations of CR, Zuojing similar formula, alkaloid fraction and non-alkaloid fraction were established, and the chemical attribute was analyzed. Using principal component analysis as main statistics, taking the internal integral area as the samples, statistical result indicated that the differences of HPLC fingerprint of different samples of preparations of CR mainly reflected on the change of content of chemical basis Y3 and Jateorhicine, and Zuojing similar formular mainly reflected on Evodiamine, rutacarpine,chemical basis Y7 and Y8. The result of hierarchical clustering showed that clustering of different preparations of CR correlated with the property of CMM. Fanzuojingwan significantly differed with the other similar formula.5. The confirmation of anti-Hp active essences of different chemical parts of Zuojingwan: The study on differences of property or activity of Zuojing similar formula and preparations of CR indicated that Zuojingwan and Cuzhihanglian had the best anti-Hp activities. The study also showed that the alkloid fraction of Zuojingwan was the main anti-Hp active substance.6. Through study on preparations of CR and Zuojing similar formula, a new mode and method of active essence screening of heat cleaning CMM and prescription compatibility based on biothermodynamics were preliminarily established.The active essence which correlated with function and indication of CMM or prescription compatibility can be explored through the study of biothermodynamics, chemical basis and pharmacological activity combined with statistical method of principal component analysis and canonical correlation analysis.7. The pharmaceutic study on Zuojing sustained floating tablets(ZJSFT): The paper tried to preliminarily explore the preparational technic of ZJSFT based on understanding the anti-Hp active fraction of Zuojingwan. The result indicated that D101 macroporous resin can concentrate and purify the Zuojing alkaloid fraction; The ZJSFT which with best floating performance and qualified cumulative drug release as well can be prepared by optimizing the technic of floating performance first and then adjusting the cumulative drug release. The drug release mechanism was joint actions of drug diffusion, bulk erosion and chalasia. It showed that release condition had no obvious effect on floating performance and cumulative drug release.