| Objective:To explore the possibility of pathogenesis of spleen deficiency after cerebral ischemia injury by literature investigation.And to explore the protective effect of invigorating the spleen andtonifying the earth formula(JPBTF)on cerebral ischemia injury and its mechanism of intervention on extracellular matrix and integrin(INT)-focal adhesion kinase(FAK)signal pathway from in vivo and in vitro,to proof by contradiction that pathogenesis of spleen deficiency after cerebral ischemia injury is possible,and to provide a new therapeutic method and entry point for clinical anti cerebral ischemia injury.Methods:1.Based on the literature review,we searched the relevant journals and literatures on acute stage of stroke in the past 10 years,and used frequency analysis to explore the distribution of syndrome elements in ischemic stroke.2.Taking the famous doctors' experience of stroke treatment included in "ancient and modern famous doctors' Golden Mirror in clinic-stroke volume" as the research object,frequency analysis and cluster analysis were used to explore the rule of medication for ischemic stroke.3.Based on the thought of microcosmic syndrome differentiation and analogy,this paper compares the similarity betweem the microcosmic structure of ECM and the functional of"spleen earth" of traditional Chinese medicine,and puts forward the treatment method of strengthening spleen and tonifying the earth to protect ECM against cerebral ischemia injury.In vivo experiments,the rats models of cerebral ischemia-reperfusion were established by middle cerebral artery occlusion.The rats were divided into sham operation group,model group,positive control group,small dosed JPBTF group,medium dosed JPBTF group and high dosed JPBTF group.The neurological deficit score,pathological morphology,blood-brain barrier electron microscopy and blood-brain barrier permeability were measured,ECM,caspase-3 were detected by immunohistochemistry,volume fraction of collagen(CVF)was detected by Masson staining,MMP were detected by RT-PCR,the int?3,p-fak,p-Akt and bcl-2 protein were detected by Western blot,neuron apoptosis was detected by TUN EL.In cell test,INT,ILK and FAK was detected by Western blot and immunocytochemistry.Results:1.There are 13 common syndrome types in the acute stage of stroke.Among which there are 7 syndrome types related to "phlegm",the cumulative frequency accounts for 53.8%;there are 5 syndrome types related to "blood stasis",the cumulative frequency accounts for 50.2%;there are 3 syndrome types related to "qi deficiency",The cumulative frequency accounts for 22.9%.2.In the acute stage of stroke,the top three syndrome key factors of disease's location in frequency are brain,spleen and liver,the top three syndrome key factors of disease's nature are phlegm,blood stasis and Qi deficiency.3.The top 10 single drugs are paeony,Achyranthes bidentata,angelica,Pinellia ternata and tuckahoe,Acorus calamus,Arisaema,Rhynchophylla,Astragalus membranaceus,Prepared Radix Rehmanniae in stroke prescriptions.4.The top 5 drugs types were tonic medicine,liver calming and wind calming medicine,blood activating and stasis removing medicine,heat clearing medicine and phlegm resolving medicine.5.Cluster analysis of high-frequency use drugs can be divided into 6 categories:the addition and subtraction of Ditan Decoction;the addition and subtraction of Lingjiao Gouteng Decoction;the addition and subtraction of Tianma,wolfberry and chrysanthemum Decoction;the addition and subtraction of Zhenganxifeng Decoction;the addition and subtraction of Liuwei Dihuang Pill;is the addition and subtraction of Buyang Huanwu Decoction.6.The different doses of JPBTF could improve the neurological symptoms of cerebral ischemia-reperfusion rats(P<0.01),improve the pathological morphology of brain tissue;The The high dosed JPBTF and the middle dosed JPBTF could protect the integrity of blood-brain barrier,reduce the permeability of blood-brain barrier,reduce the exudation of Evans blue in brain tissue(P<0.05);The different doses of JPBTF could reduce the apoptosis of nerve cells(P<0.05-0.01),The high dosed JPBTF and the middle dosed JPBTF could reduce the degradation and loss of Col IV,LN and FN in brain tissue(P<0.05-0.01);The different doses of JPBTF could inhibit the expression of MMP-9 and MMP-2(P<0.05?0.01);The high dosed JPBTF and the middle dosed JPBTF could promote the INT-FAK signaling pathway key proteins expression such as int?3,ILK,p-FAK,p-Akt and Bcl-2(P<0.05?0.01):The different doses of JPBTF could reduce the expression of Caspase-3(P<0.05?0.01).Conclusion:1.Literature research shows that in addition to brain in syndrome key factors of disease's location,spleen ranks the first,while phlegm ranks the first and Qi deficiency ranks the third in syndrome key factors of disease's nature,respectively.The results show that dysfunction of spleen in transportation result in insufficient generation of Qi and blood,at the same time dysfunction of spleen in transportation result in the dampness brewing and phlegm generation are the important pathological links of stroke.Spleen deficiency is an important pathogenesis after cerebral ischemia.According to the data mining of "ancient and modern famous doctors'Golden Mirror in clinic-stroke volume",it shows that invigorating spleen and benefiting Qi is an important therapy unit in the treatment of stroke.It provides theoretical support for the treatment of stroke with the method of invigorating spleen and tonifying earth.2.The invigorating the spleen and tonifying the earth formula can improve the state of spleen deficiency,and protect the nerve function and pathological morphology of brain tissue,reduce the degradation of ECM and inhibit the apoptosis of nerve cells in rats with cerebral ischemia-reperfusion.3.The invigorating the spleen and tonifying the earth formula may reduce the brain ischemic nerve damage by protecting ECM and intervening in INT-FAK signal pathway.4.The method of invigorating the spleen and reinforcing the earth based on the idea of microcosmic syndrome differentiation and analogy--comparing ECM with "spleen earth"is feasible to protect the neurons ECM against cerebral ischemia injury. |
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