Based On Analysis Of Neurovascular Unit With Diabetic Cerebral Ischemia Model Of Rats Brain Tissue Pathological Changes

Objective:1.To establish the model of diabetic rats,observing the effect of diabetes mellitus on the neurovascular unit structure and function of rat model;2.To establish diabetic cerebral ischemia model, And what changes in a rat model of diabetic cerebral ischemia at different times on neurovascular unit structure and function.Method:1. Constructing model:intraperitoneal injection of streptozotocin (ST Z) duplicated diabetes model, the level of blood glucosedetection to determine the successful rat model of middle cerebral artery ischemia model; replicate, observed the injury of rat brain ischemia.2. Grouping:the experimental animal were randomly divided into norma 1 control group and diabetic group, The normal control group include s normal group,control group, cerebral ischemia 3d and 7d group;diab etes mellitus group also contains four groups, diabetes group, contro 1 group, diabetic cerebral ischemia 3d and 7d groups. Using the met hod of the behavioral function tests and TTC staining, detection of di fferent groups of animal behavior, and to detect the brain infarct volu me.3.Abdominal aortic blood,Detecting the expression of serum IL-1,COX-2, TNF-α ,vWF, NSE in serum by enzyme linked immunosorbent ass ay (ELISA)4.Compared the expression of rats brain tissue of E-selectin, ICAM-1, ICAM-1 and CD34 using immunohistochemistry.5.Western Blot method for detection the protein expressionof rat brain cortex E-selectin, VCAM-1, CD346.Using the transmission electron microscope, to observe the change of brain tissue in rats of each group cells inthe infarct border zone ultrastructure.Result1) compared to the normal group and diabetic group①In diabetic group,the expression of TNF-α、IL-1、COX-2、vWF、NSE was significantly increased in serum,and have a significant difference (p<0.05);the expression of E-selection、VCAM-1、ICAM-1 increased and have obvious difference in brain tissue (P<0.05),the expression of CD34 in brain tissue decreased, there were statistical significance (p<0.05).②The diabetic group of neurons the regular shape, nuclear week gap narrow, dense nucleolus, heterochromatin decreased, swelling of mitochondria in the cytoplasm, pale, fewer cristae, Golgi complex dilatation, part of rough endoplasmic reticulum shed edema.2)Composite cerebral ischemia group compared with the pure cerebral ischemia group:①neurological score:The differences among different groups were not obvious (P>0.05); Composite cerebral ischemia difference 7d group between after waking up and before treatment was obvious (p< 0.05) Other groups were statistically significant difference.②The concentration of TNF-α、IL-1 and COX-2 in the diabetic group were higher than normal group(P<0.05); Compared with the normal g roup,The cerebral ischemia rats in 3 days and 7 days showed high ex pression,and showed significant differences(P<0.05);Compared to diabet ic group, in diabetic concurrent cerebral ischemia group, the concentra tion of them were lower with significant difference in 3 days an d i n 7 days after ischemia (p<0.05)③vWF:Compared with the normal group,The cerebral ischemia rats in 3 days and 7 days and the diabetic group showed high expression, The cerebral ischemia rats in 7 days was increased obvious and had statistical significance(p<0.05);But in diabetic concurrent cerebral isc hemia groups were lower expression in 3 days an d in 7 days,when t he composite ischemia 7d expression was the lowest.④NSE:Compared with the normal group and the normal Sham-operated group,The cerebral ischemia rats in 3 days and 7 days and the dia betic group showed high expression with significant difference (p<0.0 5);Compared with the cerebral ischemia 3 d group,The expression of t he diabetic concurrent cerebral ischemia group decreased in statistical significance (p<0.05);Compared with the diabeticl group and the di abetic Sham-operated group:the concentration of them were lowe in 3 days an d in 7 days after ischemia,The 7d group decreased significant ly and had statistical significance⑤Immunohistochemical detection the expression of E- selectin、VCA M-1、ICAM-1、CD34 in the infarct border zone:The concentration of E- selectin、VCAM-1、ICAM-1、CD34 in the diabetic group were high er than normal group;The Composite ischemia results showed the conc entration of E-selectin、VCAM-1、ICAM-1 were higher in 3 days an d in 7 days after ischemia than the normal group,but compared with t he corresponding time point was lower in cerebral ischemia;The expres sion of CD34 in infarcted zone:Compared with the normal group,The expression of ischemia group and diabetic group decreased,The expressi on of composite cerebral ischemia group was higher than the diabetic group.⑥The expression of protein expression is similar to immunohistochemistr y in rat brain infarct border zone in the Rat brain cortex E-selectin, VCA M-1,CD34,and in simple, complex cerebral ischemia blood levels and diab etes groups on the rise,complex cerebral ischemia group and 3d 7d group i t decreased within E-selectin, VCAM-1. CD34 ischemia and diabetic grou p decreased its composite cerebral ischemia 3d 7d group compared with th e upward trend.⑦Changes in morphology and ultrastructure of ischemic border areas: diabetes complex cerebral ischemia compared with simple cerebral isch emia:Ischemia 3d group appear loose organizational structure, a larg e number of nerve cell death, cell edema irregular, plastic stromal cel 1 proliferation, neuronal degradation heterochromatin, mitochondria sho rter, sparse, ribosome off; vascular endothelial cell swelling, reduction of synaptic vesicles. Ischemia 7d group has been reduced edema, isc hemic border zone in the form of structurally normal neurons, gliosis significantly, endothelial cell edema. On the other hand,diabetes co mplex cerebral ischemia than the simple more serious through observa ting of their morphology injury.Conclusion1).The model of Diabetes will injure blood vessels endothelial, and a ctivation of glial cells, stimulate inflammation,injure blood vessel neur ons.2). Combined diabetes Phlegm theory and modern material found Phleg m associated doctrine having a definite link contact with endothelial damage3).In the condition of diabetic composite cerebral ischemia, explore th e protective effect of the neurovascular unit,due to diabetes leading to cerebral ischemia is caused by many factors,and the mechanism is m ore complex,we will research through a multi-level, multi-channel, mul ti-target integrated acting on the various aspects of pathology.