Study On Bioactivity Of Curcuma Kwangsiensis Polysaccharides And Immune Analysis Method Of Curcumol

In this paper, Guangxi famous-region drug Curcuma kwangsinensis S.G.Lee et C.F.Liang was taken as research object, investigating the biological activity, mechanism and anti-oxidative of Curcuma Kwangsiensis Polysaccharides (CKP) on regulation of dyslipidemia; the present paper prepares the monoclonal antibodies of Curcumol acting as one essential composition and antitumor compound in Zedoary Tumetric Oil, and deepens the analysis on immune application of those antibodies. The results as follows.Used three dose groups, CKP Ⅰ (300mg.kg-1), CKP Ⅱ (600mg.kg-1), CKPⅢ), to feed hyperlipemia rats, and lovastatin well-known was taken as one common cholesterol-lowering drug to do the positive control. After3weeks, these rats blood was collected for the content mesurement of the following items:serum triglycerides (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), apolipoprotein AI (apoAI), apolipoprotein B (apoB), glutathione peroxidase (GSHPx), super disambiguation peroxidase (SOD) and malondialdehyde (MDA). The results show that CKP has obvious improvement effect and anti-oxidative on dyslipidemia rats, as well as the effect positively correlates with dose.By the fluorescent quantitative PCR, perilipin protein and hormone sensitity lipase (HSL) as two lipogenic genes were chosen for the target genes to analyze the correlation between the improvement effects of CKP on hyperlipemia rats and transcription expression of lipogenic genes. The results indicate that the gene expression of HSL contained in adipose tissue of hyperlipemia rats in the three CKP dose groups increases, while the expression of perilipin decreases; the gene expression of TG in serum of the three CKP dose groups shows a significant negative correlation (P＜0.05) with HSL, but a significant positive correlation with perilipin (P＜0.05).The present study curcumol and succinic anhydride was used as two reactants and4-dimethyl amino pyridine (DMAP) as one catalyst to make curcumol-succinic monoester hapten, then the hapen is conjugated to the protain by the method of EDC and finally gets curcumol antigen which shows good immunogenicity and can be applied in curcumol monoclonal antibody preparation as one immunogen. This synthesis has the following advantages:mild reaction conditions, simple procedures, stable quality and yields of synthesis high to87%.The adoption of hybridoma technique makes us get the cell strain of monoclonal antibody possessing high-secretion ability, and gets the curcumol monoclonal antibody whose content is more than90%after ascites preparation and ProteinG agarose gel affinity chromatography column purification. The identification and evaluation on antibody from aspects of type, titer, sensitivity, affinity constant, cross reaction rate prove its high titer (above106), good affinity (affinity constant2.45X10-8) and strong specificity.This study preliminarily established the curcumol ELISA analysis method and conducts methodology investigation, showing that the linear relationship (0.16-llμg.ml-1), reproducibility (9.02%), precision (＜9%), plus sample recovery (10.01%), the divergency of between board(＜10%) and within plate (＜10%) meet the requirements of the immune analysis, and the minimum detection limits is78ng.ml-1. The test results of Curcuma kwangsiensis, Curcuma wenyujin and organisms sample containing curcumol have no significant difference in ELISA method and gas chromatographic method.This results has great significance to development the medicinal value of Curcuma kwangsiensis; and provide a reference for quality evaluation system of it.