| Today ischemic cerebro-vascular disease is the one of most severe diseases in China. In spite of some improvement on thrombolysis, the development of intervention on reperfusion after ischemia has been slow. Most scientists think some drugs, having different action mechanism, should be applied together to strengthen agaist damage from the reperfusion. Theraperticcocktail of treatment has been proved to be effective.According to the traditional Chinese medicine, Qi-supplying herbs and blood-circulation -activating herbs are compatible to treat stroke for many years, which has been good effect on clinic. Recently Xue Shuan Tong injection of blood-circulation-activating effect and Huang Qi injection of Qi-supplying effect are to be compatible to treat acute stroke, expressing synergy action. Through deep analysis and review of the past researches, we find total saponins are their effective components and give the hypothesis that if the two saponins are administered together, the synergy effect could be attached on anti-ischemic reperfusion.In the study, we studied purification technologic conditions of two saponins, total saponins of Astragalus (TSA) and total saponins of Panax notoginseng(PNS). It was studied of the optimized dose of the two saponins against damage of cerebral ischemia reperfusion. By the means of cerebral ischemia reperfusion models in rats and mice, the probable mechanisms were studied from cytokines expression, adhesion molecules expression, cyclooxygenase-2 expression and transcription factors activation.1 Researches on purification technologic condition of TSA and PNS.Objective:To study the conditions of purifying TSA and PNS. Method:By different ways of chosing in macroporous resins, densities of eluents, volumes of eluting menstruums, the best condition was to be made. Then the products quality was to be validated by purification tests and analysis of the HPLC atlases. Result: D101 macro-porous resin was better to purify the two saponins. The purity quotient of PNS was 87.7% and the purity quotient of TSA was 74%. Conclusion: The preparation of the two saponins was correct to be applied in the next studies.2 Researches on compatibility of PNS-TSA in anti-ischemic cerebral reperfusion damage.Objective:To study the compatibillity of PNS-TSA in anti-ischemic cerebral reperfusiondamage. Method:Rats and mice were grouped by Uniform design test or ANOV for factorisl experiment, respectively. Rats were subjected to MCAO for 2 h, injected the two saponins immediately, and then blood supply was recovered for 22 h. Then brains were taken out of and TTC staining of brains was made to determine the size of cerebral infarction. The model of cerebral ischemia in mice by ligation of vagus nerves and common carotid arteries was used in the study and edema of brain was detected. Result: PNS-TSA can decreased size of cerebral infarction and cerebral edema significantly. The optimized doses were TSA 16mg/kg, PNS 40mg/kg for rat and TSA 32mg/kg, PNS 80mg/kg for mice. In the ANOV for factorisl experiment, it was significant of each of TSA and PNS for size of cerebral infarction and cerebral edema but interaction of them was not significant. Conclusion: PNS-TSA related in a synergistic effect of anti-ischemia either on infarction of rats and brain edema of mice , which belong to addition.3 Researches on protection of PNS-TSA in anti-ischemic cerebral reperfusion damage.Objective:To study the protection of PNS-TSA in anti-ischemic cerebral reperfusion damage. Method:MCAO and CCAO models were made with rats. TTC staining of brains was made to determine the size of cerebral infarction. HE staining of brains was also performanced to observe morphological changes. The permeability test of Evans blue (EB) and cerebral edema were determined after ischemia and reperfusion.Result: 56mg/kg, 112mg/kg PNS-TSA can decreased size of cerebral infarction and cerebral edema significantly. In the vehicle group, a number of neurons appeared degeneration, necrosis or disappearance, with tissue edema and leukocyte infiltration. In 56mg/kg, 112mg/kg PNS-TSA group, the status was improved. The content of Evans blue was decreased observably in 56mg/kg, 112mg/kg PNS-TSA group,too. Conclusion: PNS-TSA can inhibit damage of cerebral ische-mic reperfusion.4 Researches on protection of PNS-TSA in anti-oxygen free radicals of brain in CCAO mice.Objective:To study the protection of PNS-TSA in anti-oxygen free radicals of brain in CCAO mice. Method: CCAO model was made with mice. Assay of many materials of the homogenate of brains was carried out, including of CAT, GSH, GSH-PX, SOD,MDA, NOS,T-AOC,NO.Result: 112mg/kg,224mg/kg, PNS-TSA can decrease MDA content significantly and increase SOD activity, significantly. 112mg/kg PNS-TSA can increase CAT activity significantly. But the other indexes were not significant. Conclusion :PNS-TSA can improve the ability of anti-oxygen free radicals in brains of CCAO mice.5 Researches on inhibition of PNS-TSA in neutrophil infiltration,adhesion molecules expression,cytokines expression and COX-2 activity of brain in MCAO rats.Objective.'To study the inhibition of PNS-TSA in neutrophil infiltration,adhesion moleculesexpression,cytokines expression and COX-2 activity of brain in MCAO rats. Method: Rats were subjected to MCAO for 2h, injected the two saponins immediately and blood supply was recovered for 22h. Then the contents of IL-1β, CINC,MMP-9, ICAM-1, PGE2 and TNF-αof brains were asssyed by ELISA, IL-6mRNA by RT-PCR, COX-2 mRNA by in situ hybridizationhistochemistry. The activities of MPO and COX-2 were assyed too.Result: 56mg/kg,11 2mg/kg PNS-TSA can decrease significantly contents of L-1β, CINC,MMP-9,ICAM-1,PGE2 and TNF-α, expression of IL-6mRNA and COX-2 mRNA , activity of COX-2. The contents of IL-1β, and MPO activity in 112 PNS-TSA group decreased obviously in contrast to PNS group(80mg/kg).In addition to these, content of TNF-α, activity of COX-2 and expression of COX-2 mRNA in 56mg/kg PNS-TSA group decreased obviously in contrast to PNS group.Conclusion:PNS-TSA can inhibit neutrophil infiltration and inflammation in brains.6 PNS-TSA inhibit NF-kB signal transduction in brains of MCAO rats.Objective:To study the effect of PNS-TSA in NF-kB signal transduction in brains of MCAO rats. Method: Rats were subjected to MCAO for 2h, injected the two saponins immediately, and then blood supply was recovered for 22 h. Then The expressions of NF-κB protein were assayed by IHC, I-κBαand Phospho-I-κBαby Western blot.ResuIt: After MCAO, the expression of NF-κB increased markedly in vehicle group, and showed nuclear translocation. 56mg/kg,112mg/kg PNS-TSA can decrease the nuclear translocation, the positive area and integral optical density of .NF-κB. Furthermore, the expression of p- I-κBαincreased markedly and I-κBαdecreased markedly in vehicle group. 56mg/kg,112mg/kg PNS-TSA can decreased markedly p- I-κBαand increased markedly I-κBα.The positive area and integral optical density of .NF-κB in 56mg/kg group decreased obviously in contrast to PNS group. Conclusion:PNS-TSA can inhibit I-κBαdegradation and NF-κB translocation in brains.In this study , we studied the effects of PNS and TS A on the NF-κB signal transduction and its acting gene expression of in rats subjected to cerebral ischemia and reperfusion, whichhas not been reported in our country up to now. The relationship of the compatibility of total saponins of Astragalus (TSA ) and total saponins of Panax notoginseng (PNS) is to be sure by the Uniform design test, and ANOV for 2×2 factorial experiment and two linear regression, which is a few in pharmacological study in traditional Chinese medicine. Therefore, our study is innovative to a certian extent.
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