Study The Anti-Inflammatory Effects And Quality Analysis Of Different Polar Parts Of Roots Of Clerodendrum Japonicum

Objective: Take the roots of Clerodendrum japonicum as the research object,its following research: 1.Study the Anti-inflammatory effects of the roots of Clerodendrum japonicum from different parts of the polar extract,which are petroleum ether,ethyl acetate fraction,butanol fraction,95% ethanol extracts,and determine the anti-inflammatory effects sites and the anti-inflammatory mechanism of effective sites.2.The HPLC fingerprints of the effective anti-inflammatory sites of roots of Clerodendrum japonicum will be established.3.To establish method for the determination of caffeic acid,verbascoside and kaempferol in the three different polar parts of the roots of Clerodendrum japonicum.Methods: 1.Dimethlbenzene-included auricle edema in mice,carrageenan-induced the paw edema in mice,capillary permeability induced by acetic acid and the Granuloma in mice were respectively performed to research the effects of petroleum ether,ethyl acetate fraction,butanol fraction,95% ethanol extracts of the roots of Clerodendrum japonicum on anti-inflammatory,and then determine the site of effective of anti-inflammatory effects.The models of carrageenin induced paw edema were used to investigate the relationship between the anti-inflammatory effect and hypothalamus-pituitary-adrenal axis(HPAA).The rat model of carrageenan-induced acute pleurisy was also selected to observe the anti-inflamatory effect.The anti-inflammatory mechanism was investigated by detecting the contents of MDA,NO and SOD activity.The rat model of adjuvant-induced arthritis was investigated by detecting the contents of prostaglandins(PGE2),tumor necrosis factor-alpha(TNF-?),inter-leukin-1?(IL-1?),inter-leukin-6(IL-6),leukotriene-B4 in the serum.2.Using the HPLC chromatography establishing HPLC fingerprints of the effective anti-inflammatory sites of the roots of Clerodendrum japonicum.3.Simultaneous determination of the contents of caffeic acid,verbascoside and kaempferolin three different polar parts of the roots of Clerodendrum japonicum by HPLC method using Thermo ODS-2 HYPERSIL column(4.6mm×250mm,5?m),methanol-0.05% acetic acid as the mobile phase with gradient elution,flow rate of 1.0m L/min,the column temperature was 30?,detection wavelength 320 nm,and the injection volume was 15?L.Results: 1.Effective parts of petroleum ether,ethyl acetate extract,n-butanol extract,95% ethanol extract sites of the roots of Clerodendrum japonicum have inhibited the four inflammation models from the method one.Effective parts of petroleum ether,n-butanol extract of the roots of Clerodendrum japonicum have inhibited the mice ear swelling was induced by dimethylbenzene removal of the adrenal glands,the adrenal glands of mice induced by carrageenan-induced the paw edema in mice.The different polar parts of the roots of Clerodendrum japonicum could reduce the content of MDA and NO,increase the activity of SOD,and reduce contents of TNF-?,PGE2,IL-1?,IL-6,and LTB4 in serum of AA rats.2.Using the HPLC chromatography and using a gradient elution program,calculate the similarity of 10 batches of samples and standards by fingerprint similarity map software is above 0.89,the establishment the fingerprints of the anti-inflammatory effective parts ofthe roots of Clerodendrum japonicum.There are certain differences in the common peaks and peaks of the fingerprint.3.Simultaneous determination of the contents of caffeic acid,verbascoside and kaempferolin three different polar parts of the roots of Clerodendrum japonicum by HPLC method is reliable,which showed good linear relationships within their own ranges(r> 0.9996),the average recovery rate is greater than 95%with the RSD values are less than 3%.Conclusion:The effective anti-inflammation sites of the roots of Clerodendrum japonicum are petroleum ether,ethyl acetate extract,n-butanol extract,95% ethanol extract,its anti-inflammatory effect doesn't depend on HPAA system,and its associated with decreasing of MDA,NO,TNF-?,PGE2,IL-1?,IL-6,LTB4 and increase SOD activity.Establishing the fingerprinting method of the anti-inflammatory effective part of the the roots of Clerodendrum japonicum can qualitatively and quantitatively determine the characteristic components in the different polar parts of the roots of Clerodendrum japonicum.To provides scientific basic to quality method evaluation and reasonable use resources of Clerodendrum japonicum(Thunb.)Sweet.,and to provide basic research to guid clinical drug use and develop new anti-inflammatory drugs.