| Purpose: Total Saponins of Liriope Spicata Lour(TSL),extracted with macroporous resin from the roots of Liriope Spicata Lour from HuBei province, is quantitatively determined by spectrophotometry. Previous studies showed that TSL has the function of antiarrhythmia and anti-cardiac ischemia.In this study we aim to investigate the neuroprotective effect of TSL on cerebral ischemia injury and further to exploit its possible mechanism.Methods一. The extraction and refinement of TSL is quantitatively measured by spectrophotometry.二. Chemical reagent FeCL3 was locally applied on MCA to cause thrombosis.The followings are observed:the neurological deficit score;brain infarct area;the expression of nNOS immunoreactive neurons;the pathological changes of brain tissue.三. Complete cerebral ischemia reperfusion model was induced by repeated ligation and release of both common carotid artery.The SOD activity of each group was observed.四. The coagulation time with capillary method and bleeding time with tail breaking method were measured.五. The effect of TSL on RBC in rabbit was observed with hemolysis test. Results一. TSL was effectively extracted with macroporous resin under the selected experimental conditions,It showed good linear relation within 0.004mg/ml—0.02mg/ml.二. Effects of TSL on focal cerebral ischemia in rats.1. neurological deficit score (1)The neurological deficit score in TSL 11mg/kg,44mg/kg and heparin group is significantly lower (5.25±0.89,p<0.05;3.75±1.04, p<0.01;1.50±0.72,p<0.01) than the negative control group(NS group).(2)Significant difference also exists between TSL groups.2. Cerebral infarct area(1)TSLSL 11mg/kg,44mg/kg and heparin group reduced the infarct area by 43.68%(p<0.01) ,80.59%(p<0.01) and 88.99%(p<0.01) respectively.(2)Significant difference also exists between TSL groups.3. The expression of nNOS immunoreactive neurons at the border of infarct area.(1)The expression of nNOS immunoreactive neurons in NS control group and drug treated groups is apparently upregulated in comparison with sham operation group.(P<0.01).(2)The expression of nNOS immunoreactive neurons in TSL11mg/kg,44mg/kg group was downregulated by 11.57%(P<0.01) and 28.82%(P<0.01) respectively.(3)Significant difference also exists between TSL groups.4. Pathological changes in cerebral tissueSerious damage took place in NS group ,while alleviated in TSL groups ,compared with sham group.三. The effect of TSL on SOD activity after complete cerebral ischemia reperfusion in mice.1. The SOD activity in NS group dropped in comparison with sham group.2. No marked difference was found about the SOD activity among TSL groups and NS group,while it was in nimodipine group(p<0.01).四. The effect of TSL on the coagulation time and bleeding time in mice1. coagulation time(1)TSL22mg/kg,66mg/kg and heparin group prolonged the coagulation time significantly (37.25%, P<0.05; 68.75%, p<0.01; 1878.12%, p<0.01) compared with NS group.(2)Significant difference also exists between TSL groups.2. bleeding time(1)TSL 22mg/kg,66mg/kg and heparin group prolonged the bleeding time(42.19%P<0.05;73.84%,P<0.01;161.18%,P<0.01).(2)Signaficant difference also exists between TSL groups.五. The effect of TSL on RBC in rabbitsIt has no hemolysis effect on RBC.conclusionsThe extraction of TSL with macroporous resin is relatively easy and quick while the quantitative measurement of it with spectrophotometry is accurate.TSL injection could reduce the infarct area,alleviate neurological deficit,and downregulate the expression of nNOS at the border of infarct area in focal cerebral ischemia in rats;It also prolonged the coagulation time and bleeding time in mice and had no hemolysis effect. |
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