Experimental Study And Clinical Evaluation Of The Effect Of Naomaitong In Regulating CAMP,VEGF To Promote Angiogenesis After Cerebral I/R In Aged Rat

Background:Cerebrovascular disease(cerebrovascular diseases,CVD)is a common severe disease,has high morbidity,mortality,morbidity and recurrence rate.Of ischemic cerebrovascular disease(ischemic cerebrovascular diseases,ICVD)accounted for about 60%?80%of all cerebrovascular disease,which have various etiology complex,fast,symptoms,and a serious threat to the life and health of the China's elderly.So to explore a more effective treatment is valuable.Since the Angiogenesis(Angiogenesis)was applied in tumor treatment,the method is quickly to follow to the study of vascular disease,including ischemic cerebrovascular disease(ICVD).The study found that in recent years,some signal transduction pathways activated the cascade can promote the extension of new blood vessels and the establishment of collateral circulation.Among them,the cAMP/PKA signal transduction pathway was found to be a close relationship with the VEGF/VEGFR system,and effective participation in the microvascular formation after cerebral ischemia.In this paper,by the animal experiment and clinical curative effect observation of two parts.Of the animal experiments through the establishment of rat focal middle cerebral artery embolism model(middle cerebral artery occlusion,MOCA),different time points and observe the degree of nerve dysfunction in rats,the cerebral cortex ischemic area surrounding the trypan-blue and pathological features,cAMP,and positive expression of VEGF changes to study microvascular generation mechanism after ischemia rats.At the same time,through the observation in aged rats with Naomaitong through particle intervened central cAMP system and dynamic changes of VEGF to explore the mechanism of Naomaitong.Clinical observation of different times by comparing to the hospital patients NIHSS scale,TCM symptoms rating scale to evaluate the clinical efficacy of Naomaitong.Objective:1 From the young and the elderly in ischemia/reperfusion rats in different time points(I3h I/R24h,3 d,6 d,12 d)nerve dysfunction and brain tissue pathological characteristics of observation,discussion of ischemia rats after injury and mechanisms of microvascular generation;2 From tthe young model group,the aging model group,the Nimodipine group and the Naomaitong group through different time points in the cerebral ischemia/reperfusion(I/R24h I3h,12 d),3 d,6 d,cAMP and VEGF positive expression of contrast,analysis the law of the cAMP and VEGF expression changes and pulse through the mechanism of action of the brain;3 For all into the group of patients from the baseline period(day 0),treatment for 14 days,28 days NIHSS scale and grade of TCM symptoms(qi deficiency blood stasis)rating scale,reveal Naomaitong on neural function in patients with cerebral ischemia,the influence of blood running status.Methods:1 Choose Longa8 likert scale method,were compared relatively young and old in ischemia/reperfusion rats in different time points(I3h I/R24h,3 d,6 d,12 d)nerve dysfunction and pathological changes;2 Observed with immunohistochemical method and contrast comparison the young model group,the aging model group,the Nimodipine group and the Naomaitong group through different time points in the cerebral ischemia/reperfusion(I/R24h I3h,12 d),3 d,6 d,cAMP and the positive expression of VEGF;3 Choose the NIHSS scale and the grade of TCM symptoms rating scale to assess baseline period(day 0),treatment for 14 days,28 days progress situation.Results:1 The improvement of Naomaitong of aging rats nerve dysfunction score improved after ischemia.Score results showed that Nimodipine group compared with the aged model group,I/R 24 h can nerve function obstacle score lower(P<0.05),the Naomaitong group I/R 24 h,3 d lower(P<0.05).The Naomaitong group compared with the Nimodipine group,I/R 24 h and 3d,6 d,12 d are reduced.2 The improvemen of Naomaitong through an aged rats after ischemia brain tissue pathological structure.Under the light of ICVD and surrounding tissue ischemia rats central half dark belt for pathological observation,can be found in three hours after ischemia tissue edema began to emerge,microvascular around neurons and a gradual increase(photo 8).When I/R 24 h microscopically neurons and glial cells necrotic degeneration,microvascular structure damage,light microscopy showed obvious ischemia central necrosis area structure,capillaries,neurons clearance from I/R 3 h significantly expand(photo 9).I/R 6 d microvascular,neurons appear narrow gap trend of ischemia central structure fuzzy(photo 11);When I/R12d microscopically neurons,the number of nerve fibers decreased significantly,the lesions of central necrosis liquefied stove gradually formed photos(photo 12).Compared with the young model group,the aging model group ischemic brain tissue pathological damage progress faster,damage level is higher,edema,subside speed is relatively slow;Compared with pathological damage degree aging model group,the Naomaitong group and the Nimodipine group were lighter,edema,restore fast,cerebral vein through group effect is more obvious(photo 14 to 24).3 The affect of Naomaitong about through against older ischemia rats afterbrain tissue cAMP expression level.Compare with the SO group of youth,youth each point in time I/R group were reduced,the youth I/R group I 3 h,24 h I/R,3 d,6 d,12 d of the cAMP,the positive level were reduced(P<0.05,P<0.01).Compare with the SO group of older,older each point in time I/R group were reduced,older I/R group I 3 h,24 h I/R,3 d,6 d,12 d cAMP positive expression were lower(P<0.05,P<0.01).Older I/R group cAMP,the positive level as a whole with the extension of ischemia time and depressed,and I/R 3 d after reaching valley value gradually rising slowly,when I/R6 d,12 d rose significantly(P<0.05).Each moment than older I/R group,the NImodipine group I 3 h,24 h I/R,3 d,6 d,12 d cAMP positive expression were higher(P<0.05,P<0.01).Than older each moment I/R group,the Naomaitong group I 3 h and 24 h I/R,3 d,6 d,12 d cAMP positive expression were higher(P<0.05,P<0.01).Than the Nimodipine group of each moment,the Naomaitong group I 3 h and 24 h I/R,3 d,6 d,12 d of the cAMP,the positive expression were higher(P<0.05,P<0.01).4 The affect of Naomaitong about through against older ischemia rats afterbrain tissue VEGF expression level.Compared with the SO group of youth,all the positive moment VEGF levels of youth I/R group rise,youth I/R group I 3 h,24 h I/R,3 d,6 d,12 d of positive VEGF levels were increased(P<0.05,P<0.01).Compared with the SO group older,the level of VEGF positive I/R group each time has the obvious rise,aged model group I 3 h,24 h I/R,3 d,6 d,12 d of VEGF positive expression were increased(P<0.05,P<0.01).Youth I/R group of VEGF positive levels increased gradually with the extension of ischemia time,and I/R 3 d after reaching the highest maintain a higher degree.Older I/R group level of VEGF positive with ischemia time gradually increased,and the I/R 24 h after reaching the highest maintain a higher degree.Each moment than older I/R group,the nimodipine group I 3 h,24 h I/R,3 d,6 d,12 d of VEGF positive expression were higher(P<0.05,P<0.01).Than older each moment I/R group,the Naomaitong group I 3 h and 24 h I/R,3 d,6 d,12 d of VEGF positive expression were higher(P<0.05,P<0.01).Than the Nimodipine group of each moment,the Naomaitong group I 3 h and 24 h I/R,3 d,6 d,12 d of VEGF positive expression were higher(P<0.05,P<0.01).5 The adjustment of Naomaitong through nerve dysfunction in patients with senile ICVD,qi and blood of traditional Chinese medicine?The clinical efficacy evaluation results suggest:compared with the conventional treatment group,NIHSS scale score of the Naomaitong group in 14 days and 28 days is lower(P<0.01).Compared with the conventional treatment group,TCM symptoms rating scale score of the Naomaitong group in 14 days and 28 days is lower(P<0.01).Conclusion1 Due to age factors of the central nervous system degenerative changes,the degree of pathological damage after cerebral ischemia in aged rats compared with young rats is serious,and a slow speed of recovery.Naomaitong can obviously reduce the aging degree of pathological damage after ischemia rats,and promote the brain microvascular generation,improve energy metabolism disorders;2 The positive expression of cAMP can be found in the aging rats after ischemia,it decreases with time in the early ischemic(I3h-24 h)rapidly,there was a valley in I 24 h,then it became a slow recovery.The characteristics of cerebral ischemic hypoxia caused by energy depletion,acidosis may be caused.Cerebral vein access by enhancing cAMP in the expression of ischemia/reperfusion in different sites,promoting the formation of cerebral ischemia in the hindbrain capillaries,improve energy metabolism disorder.3 The expression of VEGF in normal brain tissue is found in the whole brain but less visible.After cerebral ischemia,that enhance VEGF positive expression can be found in the peripheral ischemia central area,I3h-24 h increased gradually over time,there was a peak in the I 3 d,after slow down.The characteristics for hypoxia inducing factor(HIF)VEGF caused a large number of generated.Cerebral vein access by increasing the expression of VEGF in ischemia/reperfusion different loci,promote the formation of cerebral ischemia in the hindbrain capillaries.