| Objective:1.Collating the related literature of syndromes in recovery period of ischemic apoplexy to summarize the relationship between syndromes in convalescent period,which the evolution of its etiology and pathogenesis can be understand,and provide some basis for further experimental research.2.Observing the effect of Invigorating Spleen Formula recipe on apoptosis of primary cortical nerve cells of fetal rats after hypoxia injury in vitro and the expression of apoptosis correlation Bax/Bcl-2/MEK/Erk2,which are downstream of INT-FAK pathway,and to explore the mechanism of intervention of Invigorating Spleen Formula recipe on neuronal apoptosis.Methods:1.Using Correlation Analysis to analyse correlation analysis of syndromes in the recovery period of ischemic stroke based on Literature study.Retrieving the relevant documents of CBM,CNKI,VIP from 2007 to 2017.The association rules of syndromes in recovery period of ischemic stroke were analyzed by SQL Server 2012 MAddin data mining software and Apriori algorithm,and to summarize the common syndromes and the correlation between syndromes in recovery period.2.The primary neurons of fetal rat were isolated from SD rats on gestational day 17 and cultured in six-hole plate,dividing them into(A)normal serum control group,(B)hypoxia injury model group,(C)hypoxia + nerve growth factor group,(D)hypoxia +Invigorating Spleen Formula recipe serum group,(E)hypoxia +Invigorating Spleen Formula recipe serum+ FAK inhibitor group,using Hypoxia / reoxygenation of incubator induces the latter four groups of specimens neurons to form a model of cerebral ischemia.The apoptosis of neurons was detected by flow cytometry,the expression of m RNA of Bax/Bcl-2/MEK/Erk2 was detected by RT-q PCR,and The expression of protein of Bax/Bcl-2/MEK/Erk2 was detected by was Western blot and immunocytochemistry.Results:1.Correlation analysis showed that there were 5 common site syndrome elements of ischemic apoplexy in convalescent stage,which were brain,heart,spleen,kidney and liver,and four common syndrome factor were deficiency of Qi deficiency,blood stasis,phlegm turbidity,Yin deficiency.(A)The correlation between brain and heart was the highest,followed by brain and kidney,brain and spleen,heart and kidney.(B)The relationship between qi deficiency and blood stasis was the highest,followed by blood stasis and phlegm turbidity.Qi deficiency phlegm turbidity and blood stasis,qi deficiency and phlegm turbidity,Yin deficiency and blood stasis.2.In vitro experiments:(A)Apoptosis rate of neurons in vitro: compared with normal serum control group,the apoptosis rate of hypoxic injury model group was significantly increased(P<0.05).Compared with hypoxic injury model group,hypoxia + nerve growth factor group and hypoxia +Invigorating Spleen Formula recipe serum group decreased significantly(P<0.05).Compared with the hypoxic + nerve growth factor group,the invigorating spleen and soil group decreased,the difference was not significant(P>0.05).Compared with hypoxia+Invigorating Spleen Formula recipe serum group,the apoptosis rate of neurons in hypoxia+Invigorating Spleen Formula recipe serum+ FAK inhibitor group was significantly higher,and the difference was statistically significant(P<0.05).(B)The expression of m RNA and protein of Bax/Bcl-2/MEK/Erk2: compared with the normal serum control group,the expression of m RNA and protein of Bax/MEK/Erk2 increased significantly in the model group of hypoxia injury,and Bcl-2 decreased significantly,The difference were statistically significant(P<0.01).Compared with the hypoxic injury model group,the expression of m RNA and protein of Bcl-2 in the hypoxia+ nerve growth factor group and hypoxia+Invigorating Spleen Formula recipe serum group was significantly increased,the expression of Bax/MEK/Erk2 decreased significantly(P<0.05).Compared with hypoxic + nerve growth factor group,the expression of m RNA and protein of Bcl-2 in the hypoxia +Invigorating Spleen Formula recipe serum group was slightly higher(P>0.05).Compared with the hypoxia +Invigorating Spleen Formula recipe serum group,the expression of Bax/MEK/Erk2 in the hypoxia +Invigorating Spleen Formula recipe serum+ FAK inhibitor groupwas significantly increased and the expression of Bcl-2 was significantly decreased(P<0.05).Conclusion:1.By sorting out the literatures of the recovery period of ischemic stroke and using the method of association analysis of data mining,it is revealed that the distribution of TCM syndromes in the recovery period of ischemic cerebral apoplexy has certain regularity,and the syndromes are mainly located in the brain,heart,spleen and kidney.The main factors of disease syndrome are qi deficiency,blood stasis,phlegm turbidity,yin deficiency,and the disease nature of each disease position is related to each other.With the evolution of the disease,it is mixed or transformed with each other.Its law more clearly shows the characteristics of syndromes and the evolution of pathogenesis in the recovery period of ischemic cerebral apoplexy,and the finds provide some literature support for Invigorating Spleen Formula recipe of treating ischemic apoplexy from etiology and pathogenesis that strongly related between "brain" and "spleen",and the characteristics of the relationship between "Qi deficiency","Blood stasis" and "phlegm turbidity" and the function of spleen.2.By simulating the cerebral ischemia/reperfusion injury model after neuronal hypoxic /reoxygenation injury in vitro,we studied the expression of Invigorating Spleen Formula recipe to Neuron apoptosis and the m RNA and protein of Bax/Bcl-2/MEK/Erk2 and they under the condition of the specific gene FAK was blocked.The conclusions are as follows:(A)Invigorating Spleen Formula recipe can reduce neuronal apoptosis in vitro.(B)The mechanism may be by activating the INT-FAK pathway,upregulating the expression of FAK,and further cascade reaction through the Raf/MEK/ERK kinase signaling pathway,which inhibits the expression of apoptosis correlation Bax/MEK/Erk2.and increase Bcl-2,so as to inhibit neuronal apoptosis and protect the brain from ischemic injury. |
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