Bioactivy Of Huanglian&its Active Ingredients Berberin

Huanglian (Coptidis rhizoma) is a crude drug which derived from Ranunculaceae.Huanglian is an important traditional medicine that can remove damp-heat, fire andcounteract toxicity. Because of containing multiple active ingredients and has complexconstituents, Huanglian shows a multi-target characteristic in the process of cure thedisease. The mult-target effect presented by multiple ingredients as well as probablysingle ingredient endows Huanglian with a wide range of biological activities such asanti-inflammatory, antiviral, anticancer and so on. However, it remains a difficulty thathow to use modern language to explain the theory of the overall treatment of Huanglianand its active component, as well as the mechanism of their biological activity.In this dissertation, we focuss on Huanglian and its main ingredients (berberine).The interaction between berberin and DNA&protein were studied by fluorescencetechniques and UV-Vis absorption technique, combining with comparing theantibacterial property between berberin, Huanglian and Huanglianjiedutang. The resultsprovide guidance and theoretical basis to investigate the the theory and trait of theoverall treatment of Huanglian. The dissertation includes five chapters, the maincontents and results are summarized as follows:Chapter1: The development and the actuality of the Chinese Traditional Medicineespecially Huanglian and its active ingredients (berberine) were briefly introduced. Theinteraction of active small molecules with DNA and protein were summarized; thestructures and physiologial funtions of DNA, serum albumin and hemoglobin, inaddition of the methods were described. The importance of investigation the interactionof active small molecules with DNA and protein were described, and the grounds forthe arguments of studying the bioactivity of Huanglian and the active ingredients werecarried out.Chapter2: UV-Vis absorption spectroscopy and fluorescence spectroscopy wereemployed to investigate the interaction between berberin and herring sperm DNA underthe approximatively physiological conditions. The fluorescence features and theinteraction mechanism of berberin binding to DNA were discussed detailedly, withinneutral red and acridine orange as fluorescent probes; The effects of temperature, ionicstrength, chemical denaturants, thermal denaturation and pH were studied to show the factors of the interaction, and provided further support for the binding mode; thecharacteristics of berberin binding to DNA were described; the binding constants,binding sites and thermodynamic parameters were calculated. The results provideimportant information and basis for developing new efficient drugs targeted to DNA,and designing new efficient probes for the fluorometric detection of DNA.Chapter3: UV-Vis absorption spectroscopy in combination with fluorescencespectroscopy were employed to investigate the application in the actual research ofusing berberin as a probe. The competitive binding between fluorescent probe(Ethidium bromide, Acridine orange and Berberine) and curcumin for DNA wereinvestigated to support the binding mode of berberin and DNA; the effect of anionicquenching agent, chemical denaturants, hyperthermia and pH were discussed in detail.The research results throw light on the prospective probability of using berberin as anefficient DNA probe.Chapter4: spectroscopic method was employed to investigate the interaction ofberberin with bovine hemoglobin. The binding constants, binding sites, thermodynamicparameters and binding modes were calculated and discussed; Synchronous fluorescencespectrum were used to investigate the structural change of hemoglobin molecules withthe addition of berberin; The effect of common ions on the investigated systems werestudied. The research results in our work provided basic parameters on molecular levelfor expounding the mechanism of berberin act in vivo.Chapter5: Heating circumfluence method were employed to extract berberin fromHuanglian, and the effects of operation conditions, such as ethanol concentration,backflow time, ratio of solid to liquid and extraction times were discussed; Theinteraction of bovine hemoglobin with berberin, Huanglian and Huanglianjiedutangwere compared for the first time, in addition of their anti-bacterial activity usingEscherichia coli (E. coli) as a target; The contribution of berberin to Huanglian andHuanglianjiedutang were described, combining with a certain level of pharmacologyand bioactive index. The research results provide useful lessons for compatibility andrational use of the Chinese Traditional Medicine.