RESUMO
OB JECTIVE To study the protective effects of saponins from Gleditsia sinensis on ischemic stroke with phlegm and blood stasis （ISPBS）model rats ，and to explore its mechanism. METHODS Totally 119 rats were randomly divided into normal group （normal saline ），sham operation group （normal saline ），model group （normal saline ），nimodipine group （positive control group ，5 mg/kg），G. sinensis saponin low-dose ，medium-dose and high-dose groups （3.21，6.42 and 12.84 mg/kg），with 17 rats in each group. Except for normal group ，other groups were all given high-fat diet+suture-occluded method to induce ISPBS model. The neurological function score ，water content of brain tissue ，pathological morphology of brain tissue ，the changes of hemorheology indexes （whole blood viscosity ， erythrocyte aggregation index ， Casson-viscosity）， four items of blood lipid [triacylglycerol （TG），total cholesterol （TC），low-density lipoprotein cholesterol （LDL-C），high-density lipoprotein cholesterol（HDL-C）] and inflammatory factors in serum and oxidative stress indexes [malondialdehyde （MDA），nitric oxide （NO），superoxide dismutase （SOD）] in brain tissue were determined or observed in rats. The protein expressions of B lymphocytoma 2（Bcl-2），Bcl-2 associated X protein （Bax）and caspase- 3 in cerebral tissue were also detected. RESULTS Compared with normal group ，the score of nerve function ，5 kinds of serum indexes （TC，TG，LDL-C，TNF-α，IL-1β）， hemorheology indexes ，the contents of MDA and NO and protein expressions of Bax and caspase- 3 in cerebral tissue were all increased significantly in model group （P＜0.01）. The levels of HDL-C and IL- 10 in serum ，SOD activity and protein expression of Bcl- 2 in cerebral tissue were decreased significantly （P＜0.01），and obvious lesions such as nuclear pyknosis and cell membrane fragmentation occurred in brain tissue. Compared with model group ，above indexes of administration groups were improved to different extents ，among which there was statistical significance in above indexes of G. sinensis saponin high-dose group （P＜ 0.01）. CONCLUSIONS Saponin from G. sinensis has a good protective effect on ISPBS model rats. Its mechanism may beassociated with reducing oxidative damage ， reducing the production of pro-inflammatory mediators and resisting neuronal apoptosis.
RESUMO
OBJECTIVE：To establish and optimize a extraction method of Fructus Gleditsiae Abnormallis ，and to analyze and identify chemical components of the extract simultaneously. METHODS ：Fructus Gleditsiae Abnormallis was extracted with CO 2 supercritical fluid extraction （SFE）method. Based on single factor tests ，using extraction yield as index ，extraction temperature ， extraction pressure and extraction time as investigation factors ，SFE technology was optimized with orthogonal test ，and validation test was performed. Chemical components in the extract were identified by GC-MS. Relative percentage of each component was calculated with area normalization method. RESULTS ：The optimal SFE extraction technology of Fructus Gleditsiae Abnormallis was extraction temperature of 60 ℃，extraction pressure of 300 MPa and pression time of 15 min. Average extraction of 3 times of validation tests was 1.73％（RSD＝1.78％，n＝3）. The 48 components in the extracts of Fructus Gleditsiae Abnormallis were identified，which accounted for 98.31％ of the total amount of the extracts. The extracts of Fructus Gleditisae Abnormalis mainly included organic acids ，accounting for 36.99％，followed by alkaloids ，accounting for 12.59％ in total. Main components were palmitic acid （16.62％），oleic acid （14.12％），N-aminotetrahydropyrrole（9.79％），2，6-dimethyloctane-1，7-dien-3-ol（5.95％）， tetrahydropyran（3.83％），vanillin（3.39％），etc. CONCLUSIONS ：SFE method of Fructus Gleditisae Abnormalis is established successfully，and the extract is mainly organic acids.