Exploration On The Mechanism Of Cerebral Pulse Dryness In Ischemic Stroke And Jianpi Yizhi Method Regulating MCAO Rats Endothelial Function Based On �Pulse Fu-organ� And �Interrelated Spleen And Pulse� Theories

Theoretical research sectionObjects:The purpose of this study is to explore the mechanism and prevention of cerebral pulse dryness change after ischemic stroke(IS)from the view of the location and physiological characteristics of cerebral pulse.By carding the theory of pulse fu-organ of Traditional Chinese Medicine,discussing the correlation of spleen and pulse,and analying the characteristics of the disease location of IS,and the causative mechanism of "dryness" of IS is put forward,to clarify the basic pathology of IS closely related to the spleen,to establish the dominant position of spleen weakening and pulse dryness in the pathological changes of IS and to summarize the theory basis that Jianpi Yizhi method can improve the dryness of the cerebral pulse by invigorating the spleen,moistening the pulse and removing the dryness,and achieve the purpose of treating IS.Methods:Card the related theory of pulse fu-organ and related spleen and pulse of Traditional Chinese Medicine,analyze the location and physiological characteristics of the cerebral pulse,use the original thinking of �Quxiang Bilei� from the theory of Internal Dryness that�both Zhuse Kuhe and Ganjin Cunjie are belonged to �Dryness�,deeply explore the internal relationship between cerebral pulse dryness change caused by spleen weakening and IS,combine the composition analysis of Jianpi Yizhi Decoction and the preliminary research foundation of my tutor's team,provide theoretical basis for the treatment of IS from the point of invigorating the spleen,moistening the pulse and removing dryness by Jianpi Yizhi method.Outcomes:1.The pulse fu-organ has the physiological characteristics of constancy and change.2.There is a deep connotation of �basing on camp gas,moist as nature,taking blood and body fluid,and regulating pulse by spleen�between spleen and pulse.3.Because of its unique location and physiological characteristics,the cerebral pulse has the local pathological mechanism of easy dryness and easy change.The dryness of cerebral pulse,dominated by spleen weakening,is the basic pathology and deep risk pathogenesis of apoplexy.Either it is the mechanism of moistening the pulse of the spleen by the Taiyin,the nourishing pulse of the spleen,and the dispersing of the spleen to master the pulse,or the modern clinical and experimental studies,they can provide sufficient basis for the treatment of IS by invigorating the spleen,moistening the pulse and removing dryness.Jianpi Yihzi Decoction contains effective components for IS,which have good effects in clinical and experimental studies.Experimental research sectionObjects:In order to observe the expression of bioactive substances of vascular endothelium,growth factors,related gene and proteins,and to explore the effect and possible mechanism of Jianpi Yizhi method on improving endothelial function and neural function of model rats,neurological impairment score,nitrate reductase method,immunohistochemistry,ELISA,q PCR and Western blot were used to MCAO rat model.Methods:Male SPF SD rats were randomly divided into Blank group,Model group,fasudil group(HF),Jianpi Yizhi group(JPYZ).Zea Longa method was used to replicate rat MCAO model,The Blank group was given standard feed and independent drinking water,and the other groups were given medication 14.4m L/kg/d,once a day,once they woke up after surgery,which lasted for 3,7 and 14 days respectively,HF group was given intraperitoneally fasudil hydrochloride suspension(14.4m L/kg/d),JPYZ group was given stomach invigorating Jianpiyizhi decoction(14.4m L/kg/d),respectively.Model group was given constant dose of normal saline.Nitrate reductase method was used to observe and analyze the content of NO in the brain of rats in each group.The expression levels of e NOS and VEGF were detected by immunohistochemistry method,and the content of VEGF was detected by ELISA.The possible mechanism of improving endothelial function of model rats by Jianpi Yizhi Decoction was explored from the perspective of endothelial function.q PCR was used to detect RhoA m RNA gene expression,and Western Blot was used to detect ROCK2 protein content.The possible regulatory mechanism of Jianpiyizhi Decoction on endothelial function of model rats was explored from RhoA/ROCK2 pathway.The neurological function of rats was evaluated by neurological impairment score,and the expression of Caspase-3 in the brain of rats in each group was detected by Western Blot.The possible mechanism of the improvement of neural function of model rats by Jianpi Yizhi Decoction was explored from the perspective of neurological function damage and cell apoptosis.Outcomes:1.Detection of NO,e NOS and VEGF: Compared with the Blank group,the levels of NO,e NOS and VEGF in the Model group were increased significantly(P<0.01);Compared with Model group,the levels of NO,e NOS and VEGF(immunohistochemical method)in HF group and JPYZ group increased significantly,with statistical significance(P<0.01).The influence trends of NO,e NOS and VEGF in HF group and JPYZ group were roughly the same,and the efficacy became better with the extension of intervention time.2.Detection of RhoA m RNA and ROCK2: Compared with the Blank group,the levels of RhoA m RNA and ROCK2 in the Model group were increased significantly(P<0.01);Compared with Model group,RhoA m RNA and ROCK2 levels in HF group and JPYZ group were significantly decreased(P<0.01).HF group and JPYZ group had inhibitory effects on RhoA and ROCK2.3.Neurologic impairment score and Caspase-3 test: Compared with the Blank group,neurologic impairment score and Caspase-3 level in the Model group were increased,and the differences were significant(P<0.01);Compared with Model group,neurological impairment score and caspase-3 level in HF group and JPYZ group were decreased,and the decrease was most significant at the time point of 14 days,with statistical significance(P<0.01).HF group and JPYZ group showed similar trends in neurological impairment score and caspase-3,showing better protective effects with the lasting of intervention.Conclusions:1.The pulse fu-organ has the physiological characteristics of constancy and change.2.There is a deep connotation of "basing on camp gas,moist as nature,taking blood and body fluid,and regulating pulse by spleen" between spleen and pulse.3.Because of its unique location and physiological characteristics,the cerebral pulse has the local pathological mechanism of easy dryness and easy change.It is associated with the imbalance between water in the kidney and fire in the heart,the imbalance between damp in the spleen and dryness in the lung,but the root cause is the disorder of damp in the spleen due to spleen weakening.The dryness of cerebral pulse,dominated by spleen weakening,is the basic pathology and deep risk pathogenesis of apoplexy.The dryness of the cerebral pulse can be considered as a pathological state of vascular endothelial dysfunction.Either moistening the pulse of spleen by Taiyin,or nourishing the pulse of the spleen by storing Ying Qi,or unblocking the pulse of the spleen by dispersing damp,is able to regulate dryness and prevent pathology for the cerebral pules.They can provide sufficient theories for the treatment of IS from the point of spleen.The medicinal components of JPYZ Decoction contain effective ingredients for IS,which are used together to be able to invigorate the spleen,moisten the pulse and remove dryness,improve endothelial dysfunction,improve the pathological state of IS cerebral pulse dryness,reduce the damage of many pathological factors,and mobilize the role of spleen to actively repair the damage of IS cerebral pulse dryness,prevent sudden wind disease,and play an overall therapeutic role for IS.4.MCAO model was successfully replicated by embolization of middle cerebral artery in SD rats.Jianpi Yizhi Decoction could effectively improve the symptoms of impaired neurological function in MCAO rats.5.Jianpi Yizhi Decoction may improve vascular endothelial dysfunction and achieve protective effect by increasing the levels of NO,e NOS and VEGF,inhibiting the activation of RhoA,ROCK2 and Caspase-3.