| ObjectiveCerebrovascular disease is the first disease of China urban and rural residents to death and disability.As for the further study of mechanism of cerebral ischemia reperfusion injury,people realize that oxidative stress and cell apoptosis and autophagy both play an important role in it.Tong Sheng Pill is an experienced prescription of Chinese medicine in the treatment of acute ischemic stroke in hospital.In this study,the effects of Tong Sheng Pill on cerebral ischemia reperfusion injury rats and related neuroprotective mechanisms were discussed from the perspectives of oxidative stress,apoptosis and autophagy reaction.MethodEstablish the rat model of Middle Cerebral Artery Occlusion/reperfusion(MCAO/ R)by line suppository.In the experiment,SPF grade SD male rats were used for 6-8 weeks and weight of 250-270 g.It was divided into six groups,namely the high,middle and low dose group of Tong Sheng Pill,the An Gong Niu Huang pill group,the sham operation group and the model group.With Tong Sheng Pill,An Gong Niu Huang pill,pure water,continuous lavage for 1 week,1 times a day,and observe the general situation of cerebral ischemia reperfusion rats,measuring weight,rats neurological function score in the 1,3,7 day,after the last delivery,put to death in the rat.Collect the blood and brain of rats.The size of cerebral ischemia in rats was measured by TTC staining,and the damage of cerebral ischemia was observed.The pathological damage of rat brain was observed by means of HE staining and Nissl staining.The content of LPO,MDA and SOD in rats were determined by biochemical kit.The expression of Bax,cyt-c and Lc-3 in the brain of rats was detected by Western blotting(WB),and the mechanism of neurologic function in rats with cerebral ischemia reperfusion was studied.Result Part ?: General observationThere is no significant difference in weight of rats before operation(P>0.05).On the 7th day after operation,compared with the sham operation group,the weight level of the model group was significantly different(P<0.01),and the weight of the high,middle,low dose group and the An Gong Niu Huang group increased in different degrees.Part ?: Neurological score observation1 days after operation,compared with the model group,the neurological score of rats in sham group was statistically significant(P<0.01).And compared with the model group,the high,middle and low dose group of Tong Sheng Pills,An Gong Niu Huang group was no significant difference(P> 0.05).7 days after operation,compared with the sham operation group,the neurological score of rats in the model group was statistically significant(P<0.01).And compared with the model group,the neurological score of rats in the high and middle dose group of Tong Sheng Pill and An Gong Niu Huang were statistically significant.Part ?:Cerebral infarction area observationCompared with the sham operation group,the area of cerebral infarction of rats in the model group was statistically significant(P<0.01).Two group of high dose(P<0.01)and the An Gong Niu Huang(P<0.01),compared with model group,the results have significant difference,but the difference between high dose and the the middle and low dose group was not significant(P > 0.05).Part?:Pathological morphology observationIn the sham operation group,HE staining show no obvious signs of ischemic damage in cerebral neurons.In the model group,the nerve cell ischemia damage was serious.The pathological changes of brain tissue in every medicine group were less than that of the model group,and there was a certain dose dependence trend in the improvement.In the Nissl staining method,the Nissl bodies in the sham group were densely arranged and the cells were clearly delineated.In the model group,there are sparse arrays of Nissl bodies,and even neurons disintegrate.The number of survival Nissl bodies in the brain tissue of theAn Gong Niu Huang group and the Tong Sheng pill group was more than that of the model group.Part?:LPO ?MDA?SOD ObservationCompared with the sham operation group,the LPO and MDA level of the model group was significantly higher(P<0.05),while the high and low dose group of LPO was statistically significant reduced(P<0.05)compared with model group compared,and the low dose group(P<0.01)and An Gong Niu Huang group(P<0.05)MDA has a considerable decline,the SOD activity of model group significantly reduced compared with sham operation group,the difference was statistically significant(P<0.01).Compared with the model group,the SOD activity of the high,low dose group and An Gong Niu Huang group was significantly higher(P<0.01).Part?:Bax?Cyt-c?LC-3 ObservationModel group compared with sham operation group,the expression of Bax,Cyt-c protein has increased,while the expression of LC-3 were significantly reduced,and each drug group of Cyt-c,Bax expression all have different degrees of cut,the expression of LC-3 are raised,including.The trend of An Gong Niu Huang group and high dose group is more obvious.ConclusionThe experimental results show that Tong Sheng can effectively reduce oxidative stress damage,while inhibiting the development of apoptosis after cerebral ischemia reperfusion happen,keep the autophagy reactivity,improve neurological function defect,and play a neuroprotective role.It provides a theoretical basis for clinical application of patients with AIS,and provides a new way for traditional Chinese medicine to treat acute and critical illness. |
PDF Full Text Request