| Objective In this research, our aims were to synthesize galactosides(GDE); the resveratrol liposome (RES-LIP) and the resveratrol liposome modified by galactoside (RES-GLIP) were respectively prepared, and their distributions in mice were investigated. We induced liver injury model in mice by injecting intraperitoneally CCl4 and detected the therapeutic effects of RES-GLIP on injuried liver, which may be the basis of clinical usage of resveratrol.Methods①Synthesis and identification of galactosides(GDE):HO - (CH2CH2O)n -C12H25 (n=3), isolated by silica gel column chromatography from surfactant polyoxyethylene lauryl ether, was reacted with tetraacetyl- galactosyl bromide to give galactosides. The chemical structure of GDE3 by 1H-NMR spectroscopy;②RES-LIP and RES-GLIP were prepared by thin layer ultrasonic technique. The preparative procedure of RES-LIP and RES-GLIP is as the following: the weighed reagents were dissolved in 30ml mixture of chloroform and methanol (the volume ratio was 1:1) in a 100ml round-bottomed flask. After removal of the solvent by rotary evaporation in vacuum, a lipid membrane was obtained on the flask wall. Subsequently, 10ml 6% mannitol was added into flask, and the mixture was shattered by ultrasonics for 20min at 0℃. the products were filtrated by 0.45μm filter membrane and separated. The formulation was optimized by orthogonal design test. Drug loading(DL) and entrapment efficiency (EE) were determined by RP-HPLC;③Establishment of HPLC method for analysis of RES concentration. 1.0ml of ethyl oxide was added into 0.1ml of serum or tissue homogenates, and the mixture was shaked and centrifuged at 6400r/min for 10min. The organic layer of mixture was dried under a stream of nitrogen at 40℃. The above operation was repeated for twice. The residue was reconstituted in 100μl methanol and 20μl supernatant was injected into the HPLC for analysis. HPLC analysis was performed using a Kromasil column (150mm×416mm, 5μm).
|
PDF Full Text Request