| ObjectiveStroke,80 percent of which is ischemic,has become the world’s second leading cause of death and the first cause of disability in adults.The treatment of stroke has brought a huge financial burden to families and society.At present,in spite of t-PA which is the blood vessel recanalization treatment methods the in clinical,the other methods such as Anti-excitability toxicity,Anti-oxidation,Anti-apoptosis,Anti-inflammatory,etc.,failed to covert to clinical application.Therefore,for the most fundamental cause of neurologic dysfunction after stroke,the search for new targets or drugs to promote the recovery of neural function has become a major scientific problem to be solved urgently.Neuranagenesis is an important way to restore the nerve function of ischemic stroke.The autophagy determines the survival and function of neural stem cells,but the mechanism is unknown.Qi deficiency blood stasis is the main TCM syndrome type of ischemic apoplexy.The clinical and experimental studies(including previous studies of this research)have confirmed the exact function of the recovery of neural function,but the mechanism is unknown.The "qi and blood-kidney essence-brain marrow" is closely related to NSCs,which is closely related to autophagy.Based on the above,this study proposed hypothesis: NSCs autophagy activity changes after cerebral ischemia,affecting the survival and function of NSCs.It also regulates NSCs autophagy and promotes nerve regeneration and affects its function.And from the overall level to MCAO rats model,after the autophagy inhibitors and inducers intervention treatment,in ethology score,Western blot and immunofluorescence test,observe autophagy activity changes after cerebral ischemia influence on nerve regeneration,and the function of Bu Yang Huan Wu Decoction,and further explore its mechanism and targets.Methods1.Reasearch on the protective effect of Buyang Huanwu Decoction on brain protection of ischemic apoplexy rats.Modle :The MCAO model made of the longa classic line suppository meathod.Group: The SD male rats were randomly divided into three groups: the sham operation group,the model group and the Buyang Huanwu Decoction group;then the three large groups continued to be divided into seven subgroups:0d、1d、3d、5d、7d、10d、12d.Detection method:TTC staining was used to detect cerebral infarction area in each group.Obejectie:The optimal time point of the brain protection was selected.2.Reasearch of Buyang Huanwu Decoction on the nerve regeneration of rats with ischemic apoplexy was also promoted.Group:The SD male rats were randomly divided into the sham operation group,the model group,the Buyang Huanwu Decoction group,the rapamycin group,the3-methyladenine group,the hydroxychloroquinethe group,the Buyang Huanwu Decoction and rapamycin group,the Buyang Huanwu Decoction and 3-methyladenine group,the Buyang Huanwu Decoction and hydroxychloroquinethe group.Detection time point: According to the best time points obtained from experiment 1.Detection method: Immunofluorescence was used to detect the expression of protein of nerve regeneration such as Brain Derived Neurotrophic Factor and Doublecortin.Obejectie: Reinforcing the Buyang Huanwu Decoction to promote nerve regeneration in ischemic stroke rats.3.The regulatory effect of Buyang Huanwu Decoction on Neural Stem Cell autophagy in Rats with Ischemic Stroke.Group:The SD male rats were randomly divided into the sham operation group,the model group,the Buyang Huanwu Decoction group,the rapamycin group,the3-methyladenine group,the hydroxychloroquinethe group,the Buyang Huanwu Decoction and rapamycin group,the Buyang Huanwu and 3-methyladenine group,the Buyang Huanwu Decoction and hydroxychloroquinethe group.Detection method: The expression of autophagy associated protein Beclin-1 and LC3-Ⅱ was detected by Western-Blot.Obejectie: The effect of Buyang Huanwu Decoction on Neural Stem Cell autophagy in Rats with Ischemic apoplexy.Results1.Protective effect of Buyang Huanwu Decoction on brain of Rats with Ischemic apoplexy:Compared with the sham operation group,the model group had obvious cerebral infarction,and the Buyang Huanwu Decoction group had less cerebral infarction than the model group.The best time point was: 5 days after ischemia and reperfusion.2.The Buyang Huanwu Decoction can promote the function of nerve regeneration.BDNF expression: compared with the model group,the Buyang Huanwu Decoction of the expression of BDNF in the rat brain was up-regulated;the rapamycin group of the expression of BDNF in was up-regulated compared with the model group;the expression of BDNF was up-regulated in the group of the Buyang Huanwu Decoction +rapamycin group compared with the Buyang Huanwu Decoction;and the expression of in the group of Buyang Huanwu Decoction+rapamycin group was up-regulated compared with the rapamycin group;the 3-methyladenine group and the hydroxychloroquinethe group compared with the model group had no obvious regulation effect;the Buyang Huanwu Decoction +3-methyladenine Group and the Buyang Huanwu Decoction +hydroxychloroquinethe group had no obvious regulation effect.DCX expression: compared with the model group,the Buyang Huanwu Decoction of the expression of DCX in the rat brain was up-regulated;the rapamycin group of the expression of DCX in was up-regulated compared with the model group;the expression of DCX was up-regulated in the group of the Buyang Huanwu Decoction + rapamycin group compared with the Buyang Huanwu Decoction;and the expression of DCX in the group of Buyang Huanwu Decoction+rapamycin group was up-regulated compared with the rapamycin group;the 3-methyladenine group and the hydroxychloroquinethe group compared with the model group had no obvious regulation effect;the Buyang Huanwu Decoction +3-methyladenine group and the Buyang Huanwu Decoction +hydroxychloroquinethegroup had no obvious regulation effect.3.Buyang Huanwu Decoction regulates neural stem cell autophagy:The expression of autophagy associated protein Beclin-1 and LC3-Ⅱwas up-regulated in the model group compared with the sham operation group;the expression of Beclin-1and LC3-Ⅱ was up-regulated in the Buyang Huanwu Decoction group compared with the model group;.the protein expression of Beclin-1 and LC3-Ⅱwas up-regulated in rapamycin group compared with model group;and the expression of Beclin-1 and LC3-Ⅱ was down-regulated in the 3-methyladenine group compared with the model group;the expression of Beclin-1 and LC3-Ⅱ protein was down-regulated in hydroxychloroquine group compared with model group;the protein expression in Buyang Huanwu Decoction group was lower than that in Buyang Huanwu Decoction+rapamycin group.Compare to the Buyang Huanwu Decoction there was no significant difference in regulation of Buyang Huanwu Decoction+3-methyladenine group and Buyang Huanwu Decoction +hydroxychloroquinethegroup group.Conclusion1.Buyang Huanwu Decoction pretreatment and follow-up treatment have protective effect on the brain of ischemic stroke rats.2.Buyang Huanwu Decoction can upregulate the expression of BDNF and DCX,which can promote nerve regeneration in rats with ischemic stroke,and suggests that the effect of Buyang Huanwu Decoction on nerve regeneration is related to its regulation on autophagy.3.Buyang Huanwu Decoction can up-regulate the expression of autophagy related protein Beclin-1 and LC3-Ⅱ and promote autophagy,which may be the main mechanism and target of promoting nerve regeneration in ischemic stroke rats.4.Buyang Huanwu Decoction may promote Cerebral nerve regeneration by regulating autophagy in Rats with Ischemic Stroke... |
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