| Objective and significance:Using NF-κB signaling pathway and NLRP3 inflammatory body axis as the starting point,the changes of target protein expression were detected by observing rat m NSS score,brain water content,infarct area,cell morphology and apoptosis.The intervention effect of Yiqi Tongmai Powder on focal CIRI inflammation was verified.Methods:（1）Experimental group:all rats were randomly divided into sham operation group,model group,Yiqi Tongmai Powder low dose group,Yiqi Tongmai Powder medium dose group,Yiqi Tongmai Powder high dose group and Ginkgo Leaf Group,with 12 rats in each group.（2）The MCAO rat model was established by wire bolting method.After 90minutes,the outlet bolt was pulled out to simulate local reperfusion.False surgery only do blood vessel separation,do not insert thread suppository.（3）With reference to the Zea-Longa five-point neural function score table,the rat model with low and too high sifting score.When the thread bolt is placed in the place where the depth is not up to standard or the bleeding is too much,it is also eliminated.（4）The ginkgo leaf group was given Ginkgo biloba leaf 680 mg kg-1、Yiqi Tongmai Powder low dose group mg kg-1、traditional Chinese medicine 290 mg kg-1、Yiqi Tongmai Powder medium dose group 540 mg kg-1、Yiqi Tongmai Powder high dose group 1080 mg kg-1,sham operation group and model group equal dose saline.（5）The morphological changes and apoptosis of nerve cells were observed by HE staining and TUNEL staining.The expression of brain tissue was measured by ELISA method and the expression of brain tissue NF-κBp65、TLR-4、NLRP3、ASC、Caspase-1 by WB method.Result:（1）the success rate of MCAO model rats was about 76.9%.（2）The expression of m NSS score,brain water content and percentage of infarct area were clearly rised in the model subset than in the sham subset（P<0.01）.Compared with the model subset,Yitongmai Powder and Ginkgo Leaf subset decreased visibly（P<0.01）.Compared with Ginkgo Leaf subset,Yitongmai Powder in middle and high dose subset than in Ginkgo Leaf subset（P<0.01）.The m NSS score of low dose group was significantly lower than that of ginkgo leaf group（P<0.01）,and there was no significant difference in the rest.（3）Histopathological changes and apoptosis of cerebral edema:compared with sham operation group,model group was distinctly enhance than sham operation group（P<0.01）.There was distinctness drop in each dose subsit and Ginkgo leaf subsit of Yiqi Tongmai Powder（P<0.05,P<0.01）,Compared with Ginkgo leaf subsit,the apoptosis of middle and high dose group was more palpable than that of depress（P<0.05）.（4）WB protein expression was detected:compared with sham operation subset,the expression of model subset was visibly heightened（P<0.05,P<0.01）.The expression of each dose subset of Yiqi Tongmai Powder was significantly depress（P<0.05,P<0.01）,and the NF-κBp65、NLRP3、ASC content of brain tissue in Ginkgo Leaf subset was distinctly decreased.Compared with ginkgo leaf group,the NF-κBp65、ASC、Caspase-1 content of Yiqi Tongmai Powder in low and medium dose group was significantly down-regulated（P<0.01）,and the NF-κBp65、ASC、Caspase-1、NLRP3 content in the high dose group of Yiqi Tongmai Powder decreased significantly（P<0.05,P<0.01）.（5）ELISA detection of protein expression:compared with sham operation subset,histone expression in model was distinctly growed（P<0.01）.The L-18content of brain tissue in low and middle dose group of Yiqi Tongmai Powder decreased significantly（P<0.01）compared with that in Ginkgo biloba group,and that in high dose group of Yiqi Tongmai Powder decreased significantly（P<0.01）.Conclusion&Significance:（1）The model of cerebral ischemia-reperfusion was successful after the model was established by the method of linear suppository.（2）Yiqi Tongmai Powder canvisibly antagonize the cascade reaction induced by inflammatory signal in CIRI,can effectively inhibit the expression of TNF-α、IL-18,improve the manifestations of nerve function injury,cerebral infarction area and brain water content in rats.It also inhibited the degeneration and apoptosis of rat brain tissue.（3）Yiqi Tongmai Powder can effectively regulate the expression of TLR4,inhibit the transcription and expression of NF-кB inflammatory signals downstream,and also interfere with the expression of NLRP3 inflammatory body axis mediated by the NF-кB.In order to prevent theinflammatory response and play a neuroprotective role.（4）Yiqi Tongmai Powder is more effective in treating cerebral ischemia-reperfusion injury than Ginkgo biloba leaves,and the curative effect is positively correlated with its concentration. |
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