| With population growth, the aging society is coming, As the incidence of the ischemic cerebrovascular disease (ICVD) is increasing year by year, it is becoming the focus on prevention and treatment. In recent years, many studies have shown that traditional Chinese medicine Ginseng, Sanqi, Dansen have therapeutic effect on incidence ICVD and experimental ICVD.Ginsenoside Rbl, Rg1, Rg3, Re and notoginsenoside R1were selected as experimental objects. Uniform design compatibility combination was adopted in6concentrations ranging from1×10-4to1×10-9mol·L-1. By antioxidant experiment in vitro, the effect on eliminating free radicals and reduction ability of the samples were observed to proceed preliminary potency exploration. By the nerve cells serum deprived model the best combination samples for the subsequent animal experiment and the further potency studies were screened using nerve cell as target. By animal pharmacodynamics study, the potency of the best combination samples was further confirmed. In order to get combination sample of efficacy significantly, and provide theoretical and experimental basis for the clinical application of composition sample of saponin monomer, for the prevention and treatment of ICVD and cerebrovascular dementia, uniform design high-throughput screening was used in this topic and the design was riched in pharmacodynamics study on traditional Chinese medicine and its compound.1The result of antioxidant experiment in vitroThe result indicated that the activity of combination samples on anti-DPPH (the highest clearance ratio was only31.7%) and reduction ability (the highest was38%) were higher than5kinds of saponin monomers in the concentration of1×10-3mol·L-1(the highest clearance ratio was27%, reduction ability was10%), far less than the positive control vitamin E(the highest clearance ratio was87%, reduction ability was89%).2The result of cytology experimentThe result indicated that the nerve cells were protected by ginsenoside Rb1, Rg1, Rg3, Re, notoginsenoside R1and vitamin E (the highest ratio of cell survival was78%). The protective effect of compatibility combination samples was stronger than the monomer and vitamin E for the nerve cells(the cell survival ratio were higher than78%, up to93%).After many times of screening, finally three combination samples of the A11, A12, P5whose survival ratio was more than90%had been gotten.3The result of animal pharmacodynamics experimentThe result of mice individual behavior experiment indicated that passive evading dysfunction of mice, shortening hypoxia survival time and mouth breathing number were caused by cerebral ischemia-reperfusion (p<0.01). Compared with solvent model group, the potency of A11, A12, P5combination samples were stronger than monomers (final concentration of1×10-5mol·L-1). The learning and memory impairment were obviously improved and the hypoxia tolerance ability were obviously enhanced by A11, A12, P5combination samples (p<0.01). There was no significant differnence at pharmacodynamic action between the combination samples.The result of mice enzymology experiment indicated that the activity of SOD in serum, brain and heart were reduced, the content of NO in serum, brain and heart, and the content of ALT、LDH in the serum were increased, and the content of LDH in brain and heart were reduced by cerebral ischemia-reperfusion (P<0.01). Compared with the solvent model group, the potency of A11, A12, P5combination samples were stronger than monomers (final concentration of1×10-5mol·L-1). The activity of superoxide dismutase was increased, the content of nitric oxide was reduced and the content stability of glutamic-pyruvic transaminase and lactic dehydrogenase were held by A11, A12, P5combination samples (P<0.01). Among the combination sample groups, A12combination samples had a stronger efficiency on the activity of SOD in the serum and the content of NO in brain (P<0.05).The research results above indicated that the uniform design compatibility used in this study was similar with the traditional Chinese medicine compatibility. Three best combination samples screened by high-throughput had significantly protective effect on cerebral ischemia and reperfusion injury. The potency of A12was the best in them. A12would be carried on further formulation studies in order to be used in clinical treatment for ICVD. The uniform design-high throughput screening technology is suitable for mass potency screening of the traditional Chinese medicine effective component, and plays an important technical support role in promoting the study on mass potency screening of the traditional Chinese medicine and its compound. |
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