| Objective:Stroke is the second major cause of human death,which was with high morbidity,high mortality,high disability rate and low prognosis rate.The pathogenesis of stroke was very complicated and we also lack of effective treatment for it.From the clinical treatment,we found that traditional Chinese medicine Fu yuan Xing nao Decoction has great cure rate and good prognosis effect about stroke.We constructed the cerebral ischemia reperfusion injury model in SD rat and oxygen glucose deprivation cells model in PC12 cells.We study to Fu yuan Xing nao Decoction and its active ingredient such as stachydrine and colchicine in cerebral protective effect.We preliminarily study the effects and possible autophagy mechanisms of stachydrine and colchicine in vitro.Methods:We constructed the ischemic reperfusion injury model by the MCAO method.The animal were randomly divided into sham group,ischemia reperfusion group,Fu yuan Xing nao Decoction low dosage group(5.5g/kg),medium dose group(11g/kg)and high dosage group(22g/kg).After 15 min ischemia,the decoction was gavaged once to rat.The stachydrine(30mg/kg)and colchicine(0.5mg/kg)was administered to rat through vena caudalis after ischemia by once.After reperfusion,the whole blood was centrifuged and serum was taken to detect the changes of SOD and MDA.ELISA was used to detect the change of TNF-α and IL-1β in the serum,and brain tissues were stained with related staining to observe brain tissue damage,pathological changes and neuronal apoptosis.TUNEL staining was used to detect apoptotic cells in rats brain tissues;HE staining and transmission electron microscope were utilized to observe the pathological damage in micro structure and ultra structure in the rats brain tissues.Results:1.Compared the model of ischemia reperfusion group to Fu yuan Xing nao Decoction low,medium and high(5.5,11,22g/kg)dosage group,we also compare to stachydrine(30mg/kg)and colchicine(0.5mg/kg)treatment group,we found that rats neural function score decreased significantly;TTC staining results showed that compared with model group the percentage of cerebral infarction of Fu yuan Xing nao Decoction treatment group was significantly decreased(P<0.05);H&E staining showed that the cell damage in brain tissue of rats in the model group,the cell gap changed,shrinkaged,the damage to the brain tissue of rats were reduced with treatment by those drugs;TUNEL staining showed that the morphology of apoptotic cells in sham group of brain cells of normal brown,compared with the model group,the treatment group were reduced the number of apoptotic cells and had statistical significance(P<0.05);NISSL staining showed that the drugs were given complete the number of nerve cells increased in comparison with the model group(P<0.05).The content changes of SOD and MDA in serum were compared with model group,the content of SOD treated groups were increased(P<0.01);the content of MDA decreased compared with the model group(P<0.05);the changes of ELISA in serum of TNF-α and IL-1β,the experimental groups were decreased compared with the model group(P<0.05).2.With MTT detection of cell survival rate,compared with OGD groups,stachydrine and colchicine treatment group cell survival rate were increased(P<0.05);stachydrine and colchicine can reduce the apoptosis induced by OGD stimulation,and reduce the expression of ROS in cells;stachydrine and colchicine can reduce TNF-α and IL-1βexpression in the m RNA level(P<0.05),compared with drug group the OGD groups in the expression of abnormal protein were improved(P<0.05).Conclusions:1.Fu Yuan Xing Nao Decoction can improve the ischemia reperfusion injury in rats caused by behavioral disorders and reduce cerebral infarction percentage,has protective effects on brain nerve cells and reduce brain injury;stachydrine and colchicine has a protective effect on cerebral ischemia reperfusion injury.2.Stachydrine and colchicine can increase the survival rate of PC12 cells after OGD stimulation,decrease cell apoptosis and the content of ROS in cells. |
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