Preparation Of Anti-endotoxin Fractions From Biota Orientalis And Paeonia Suffruticosa Andr, And Theirs Bioactivities.

Objective: Lipopolysaccharide (LPS) is an outer membrane component of Gram-negative bacteria and exhibits powerful immunostimulatory and inflammatory activities. It is composed of a polysaccharide part, O-antigen and core regions, and a lipid Anchor called lipid A. LipidA moiety is known to be essential for the activity of LPS. LPS takes an important place in sepsis, so it is effective to prevent sepsis by neutralizating and destructing LPS. Many studys indicate that many traditional Chinese medicines are natural antagonist of LPS in vitro and in vivo, but a further study is obstructed on some potential anti-LPS TCM in absent of an effective tracking approach. The aim of our study is to isolate anti-LPS components from Biota orientalis and Paeonia suffruticosa Andr that can highly bind to Samonella Lipid ARe595 in our preliminary experiment, then further study their bioactivity of anti-LPS in vitro and in vivo.Methods: (1) we established an effective tracking isolation approach by immobolisating Samonella Lipid A Re595 onto a Non-derivatised cuvett of Affinity Biosensor which was evaluated by Dissociation equilibrium constant (KD) with the ligand- polymyxin B(PMB).(2) we isolated anti-LPS fractions from Biota orientalis and Paeonia suffruticosa Andr by many means which included biosensor, solvents abstraction, polyamide chromatography, silica gel colum chromatography and IEC- HPLC. (3) we investigated anti-LPS activities of the fractions in vitro which included neutralization of LPS and inhibition of TNF-αrelease from LPS-stimulated RAW264.7, and in vivo by protection of fractions to mice challenged by LPS and the heat killed E coli.Results:(1) The resonance peak of immobolisation Samonella Lipid A Re595 onto a Non-derivatised cuvett of Affinity Biosensor was a symmetrica singlet with a 3.4ng/mm2 (1994 arc second) of coating amount and the KD was 5.598×10-8 M binding to the ligand PMB. (2) We got a fraction bingding highly to lipidA from Biota orientalis, unfortunately it neither protected mice from challenging by lethal dose LPS with 22.2% of survival nor neutralizated LPS in vivo. (3) We also got a fraction binding highly to lipidA from Paeonia...