| Objective:To investigate the effect of An Nao Ping Chong Tang on the expression of PKA,pCREB and BDNF protein in rats with cerebral hemorrhage,and to explore its mechanism of neuroprotection in rats with cerebral hemorrhage.Methods:SD rats were divided into the following groups according to the random number table method: normal group,sham operation group,model group,An Nao Ping Chong Tang low dose group,An Nao Ping Chong Tang high dose group,piracetam group,autologous The cerebral hemorrhage model was made by blood caudate nucleus injection method.Each group consisted of 4 subgroups of 6h,3d,7d and 14 d.The corresponding groups were given the corresponding drugs,1 time/d,normal group,sham operation group and model group.The same dose of distilled water was given,and then the brain tissue was decapitated at the corresponding time points.The neurological deficit scores,brain water content,HE staining were used to evaluate the changes of brain tissue morphology and Western blot.Expression of PKA,P-CREB,and BDNF in brain tissue.Result:1.Neurological deficit scores:The neurological deficit scores of the model group were higher than those of the normal group and the sham operation group(P<0.05).Compared with the normal group,the neurological deficit scores of the sham operation group did not change significantly(P>0.05).Compared with the model group,the low dose of traditional Chinese medicine Group,Chinese medicine high-dose group,piracetam group cerebral hemorrhage decreased on the 3d?7d?14d(P<0.05),high-dose Chinese medicine group compared with low-dose Chinese medicine group,piracetam group,after cerebral hemorrhage On the 7d and 14 d,it decreased(P<0.05).2.Brain water content: The water content of brain tissue in the model group was higher than that in the normal group and the sham operation group(P<0.05).Compared with the normal group,the water content in the brain tissue of the shamoperation group did not change significantly(P>0.05).Compared with the model group,the low dose group of Chinese medicine and piracetam group decreased on the3d?7d?14d of cerebral hemorrhage(P<0.05),and the high dose group of traditional Chinese medicine decreased at each time point(P<0.05).Compared with the low dose group of Chinese medicine and piracetam group,the water content of brain tissue decreased on the 7d and 14 d after cerebral hemorrhage(P<0.05).3.HE staining: Microscopic microscope observation: The normal group and the sham operation group were densely packed and arranged neatly.The neurons were rich in cytoplasm,lightly stained,the nucleus was in the middle,the nucleolus was clear,the cytoplasm was not red stained,and the intercellular space was dense and edema.Model group: visible cell necrosis,necrotic cells mainly manifested as nuclear pyknosis,vacuolization;cell edema,increased glial cells,and neuronal reduction and degeneration.Low-level,high-dose group and western medicine group showed different degrees of cell edema,nerve fiber degeneration and inflammatory cell infiltration,and degeneration and necrotic cells appeared.With the prolongation of administration time,the degree of edema and inflammation decreased gradually.On the third day of the dose group,a small amount of mild edema was found in the cells,and the inflammatory cells were decreased.The effect of the western medicine group and the low dose group was inferior to that of the high dose group.4.Western Blot test:(1)PKA: There was no significant change in the expression of PKA in the normal group and the sham operation group.Compared with the normal group and the sham operation group,the expression of PKA in the model group increased at the same time point and reached the peak at 7 days;compared with the model group.The expression of PKA was significantly decreased in the low-dose group of Chinese medicine and the piracetam group on the 7d and 14d(P<0.05),and the high-dose group of Chinese medicine began to decrease significantly on the 3d(P<0.05).The high-dose group of traditional Chinese medicine was compared with the low-dose group of Chinese medicine and piracetam group,and the expression ofPKA was down-regulated on 3d of high-dose Chinese medicine group(P<0.05).(2)pCREB: The expression of pCREB was not significantly changed in the normal group and the sham operation group.Compared with the normal group and the sham operation group,the expression of pCREB was increased at the same time point in the model group,and increased significantly on the 7d(P<0.05).In comparison,the expression of pCREB increased in the low dose group and the piracetam group at 7 d and 14 d(P<0.05),and increased in the third dose of the high dose group(P<0.05).Compared with the group,the expression of the low dose group of Chinese medicine and the piracetam group decreased on the 7d and 14d(P<0.05).(3)BDNF: There was no significant change in the expression of BDNF in the normal group and the sham operation group.Compared with the normal group and the sham operation group,the expression of BDNF increased at the same time point in the model group,and increased significantly on the 7d(P<0.05).Compared with the model group The expression of BDNF increased in the low-dose group,the high-dose group of Chinese medicine and the piracetam group at different time points.Compared with the high-dose group,the expression of BDNF decreased in the low-dose group and the piracetam group at the 7d and 14d(P< 0.05).Conclusion:1.An Nao Ping Chong Tang can improve the score of neurological deficit in rats with cerebral hemorrhage,reduce brain edema,and play a role in cranial nerve protection.2.Possible mechanism: inhibit the expression of PKA in cAMP/PKA signaling pathway,down-regulate the expression of AQP4 in brain tissue,promote the phosphorylation of CREB,induce the expression of BDNF and play a role in brain neuroprotection.3.An Nao Ping Chong Tang has a dose-effect relationship in the treatment of cerebral hemorrhage in rats. |
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