Biological Activities Of Huanglian Jiedu Decoction And Its Related Components

Traditional Chinese medicine(TCM) and TCM formula are complex systems with numerous constituents and targets acting as a whole. Due to the complexity of constituents and the mechanism of effective component in bio-system, it has become the difficulty for studying TCM and new drug development. Hence, investigating the active constituents of TCM is of great significance for deciphering the chemical basis and mechanism of TCM. Based on that, complex constituents, complex ways of action, complex targets which are characteristics of TCM should be considered in study of TCM. In this paper, providing new information to the study of thermodynamic properties of bioactive moleculesand by adopting microcalorimetry to analyze the characteristics of TCM from holistic perspective. At the same time, from the molecular level to study the effective compounds by adopting spectroscopic approaches, electrochemical methods and molecular modeling can not only illustrate the effective constituents of TCM but also have a far-reaching influence on understanding the theory of TCM and exploit the new drug from TCM. Deoxyribonucleic acid(DNA) that contains the genetic instructions for the development and functioning of living organisms is an obvious focus of attraction for many researchers. And it is the target of many drugs especially the antitumor drugs as well. Consequently, DNA is an important biological model to study the mechanism of drug action from the molecular level. In this dissertation, with Huanglian Jiedu decoction(HLJDD) and its relevant constituents as research object, by adopting spectroscopic, electrochemical and molecular modeling approaches, to explore the mechanism of interaction between jatrorrhizine, palmatine and coptisine with DNA. On the macroscopic point of view, we analyze the thermal dynamic properties of HLJDD and its relevant constituents by adopting microcalorimetry. These could provide reference and basis for the initial establishment of TCM studying model to research TCM based on biothermo-kinetics study. The dissertation is separated into five chapters and main contents are as follows: Chapter one: The development and the actuality of TCM, especially the progress of TCM formulae and active TCM components were briefly introduced. The methods and progress in studying of the interaction of active small molecules with DNA, and the constituents pharmacological study of HLJDD were summarized. It mainly elaborated the great significance of studying TCM by adopting spectroscopic, electrochemical and molecular modeling approaches and biological thermodynamics methods for illustrating the acting mechanism and material basis of TCM. Chapter two: Spectroscopic approaches(UV-Vis spectroscopy, fluorescent spectroscopy) and molecular modeling method were employed to reveal the interaction between jatrorrhizine and palmatine with hs DNA. The effects of operation temperature, chemical denaturant, thermal denaturation and p H were investigated to show the factors of the interaction. In addition to experimental approaches, molecular modeling was conducted to verify binding pattern of jatrorrhizine-DNA and palmatine-DNA. This study preliminarily established the thermodynamic studying methods for the interaction of active constituents with DNA, and provided important information and scientific basis for effectively obtaining the thermodynamic property. Chapter three: Spectroscopic approaches and electrochemical methods( cyclic voltammetry and electrochemical impedance spectroscopy) were employed to reveal the interaction between jatrorrhizine and palmatine with ct DNA. The interaction mechanisms and the effects of different factors were discussed detailedly. The experimental results verified the consistency of the spectroscopic and electrochemical methods, and provided a variety of methods for further studying of the interaction between small molecules and DNA. Chapter four: The methodology of investigating active small molecules binding with DNA has been discussed in the previous two chapters. On the basis of this, coptisine has been selectived as the research object. The results suggested that intercalation mode was the main binding mode between coptisine with ct DNA. In addition to experimental approaches, molecular modeling was used as a tool to verify and expand the experimental outcomes from the theoretical point of view. Multispectroscopic and electrochemical studies revealed that this interaction process is predominantly a dynamic process. Furthermore, the circular dichroism spectroscopy confirmed that coptisine not only interacted with the ct DNA, but it also perturbed ct DNA structure. In this chapter, the combination of spectral deciphering with electrochemical and molecular modeling methods unveiled a more comprehensive information of the interaction between coptisine with ct DNA.Chapter five: Microcalorimetry method was used to describe the bioactivity of HLJDD and its relevant constituents prescription. The inhibitive activity of three different decoction made by different extraction procedure was consistent with the clinical efficacy. The study preliminarily proved the feasibility of studying TCM from macro-perspective by microcalorimetric method. It has been established that the studying model for TCM reach, which based on microcalorimetric technique. The research results could provide reference for clinical new drug development.