The Intervention Of Tong Xinluo On Structure Of Cerebral Microvascular Of MCAO Rats

Ischemic cerebrovascular disease is one of the most intractable problems in the field of modern medicine. Its morbidity, mortality and disability rate severely threaten patients’life and quality of life, also cause huge losses to the society. However, there is no breakthrough in the treatment of the disease at present.Ischemic cerebrovascular disease includes transient ischemic attack, cerebral thrombosis, cerebral embolism, etc. Ischemic brain injury involves a series of complicated pathophysiologic changes including excitatory poisoning, damage of ionic homeostasis, depolarization of peripheral area of infarction, oxygen free radical damage, inflammatory reaction, blood-brain barrier damage and cells apoptosis. Which presents dynamic changes on. According to the distribution of arteries, ischemic brain tissue can be divided into entral ischemic area and peripheral area, cerebral blood flow in the central ischemic drops rastically. There are different degree of cerebral blood flow is reduced in ischemic peripheral zone. The farther area from central has less cerebral blood flow reduction. In the center of the cerebral infarction area, neuron necrosis occurs within several minutes. While in ischemic and half dark band in ischemia because insufficient collateral vessels can be part of the ompensation of cerebral blood flow. In recent years, under the guidance of TCM collaterals disease theory,our team gained a lot of research achievement based on " Toxin damage brain collaterals" theory proposed by academician Wang Yongyan. In order to further elaborate the orrelation between brain tissue structure, function under pathological state and brain microenvironment changes, their interaction with brain cell function, and improvimg the clinical cure rate of cerebrovascular disease and exponding the academician Wu Yiling put forward "Based on microvascular disease degeneration disease from the context to pass, rendezvous and biochemical research" are carried out in full swing. We selected this study based on brain microvascular pointcuts, using modern technology, we proved the morphology and function of TCM collaterals in the human body and innovated.Therefore, it is of rofound to explore cerebral infarction pathological mechanism and to seek effective intervention strategies and treatment measures, to clarify its mechanism of action. There is great significance to restore damaged the body function, clinical patients with reduced mortality and morbidity.Objective1. To validate the restorative effect of Tong xinluo in aspect of neural function and morphology on MCAO rats;2. To observe the intervention of Tong xinluo on structure of cerebral microvascular of MCAO rats;3. To observe the intervention of Tong xinluo on promoting and inhibiting factor in microenvironment of cerebral microvascular regeneration on MCAO rats.Method1. We evaluated PMCAO model and Tong xinluo’s effect on neural functional recovery through two behavioral test, neurological function score and gait analysis system;2. We observe general morphological structure of brain tissue and various cells through HE stain;3. Cerebral infarction volume ratio is calculated after TTC staining to evaluate MCAO model and pharmacological effect of Tong xinluo;4. ELISA and IHC were implemented to observe the amount of vascular endothelial growth factor (VEGF), angiopoietin-1(Ang-1), glial acidic protein (GFAP) fibre, endodermis chalone (ES) which are associated with environment of brain microvascular regeneration, finally revealed regeneration of cerebral microvascular after acute infarction on rats;5. To observe the effects of Tong xinluo on each link above.Result1. Nerve function score results:relative to the1day model group, Tong xinluo group and positive control medicine group two scoring ratio have no statistical difference. Relative to the3day model group, Tong xinluo group two scoring ratio have with significant difference (P<0.01). Two score ratio in the positive control medicine group has obvious difference (P<0.05). Results indicate that two neural function recovery in drug groups were better than the model group, especially in the3day.2. The gait analysis system results:1)In the Step Sequence Regularity Index:Drug delivery after1day, model group that compared with and sham group decreased, with obvious difference (P<0.05); Tong xinluo group that compared with and model group rised, with obvious difference (P<0.05). Drug delivery after3day, model group that compared with and sham group decreased, with obvious difference (P<0.05).2) In the Run Duration:Drug delivery after1day, model group that compared with and sham group rised, with obvious difference (P<0.05); Tong xinluo group that compared with and model group decreased, with obvious difference (P<0.05). Drug delivery after3day, model group that compared with and sham group rised, with extremely significant difference (P<0.001); Tong xinluo group that compared with and model group decreased, with obvious difference (P<0.05).3) In the Right Paws Print Area:Drug delivery after3day, model group that compared with and sham group decreased, with obvious difference (P<0.05); Tong xinluo group that compared with and model group rised, with obvious difference (P<0.05).3. Cerebral infarction volume percent results:by TTC staining, sham control group have not see the white infarcts in the rats brain. Model group, Tong xinluo group and positive control medicine group all have occurred different degrees of the white infarcts. Relative to the1day and3day model group, the cerebral infarction volume percent of Tong xinluo group have obvious difference (P<0.05).4. HE dyeing observation results:Observed under low power light microscopy, rats brain gray matter and white matter of sham group and sham control group clearly boundaries. Pia mater is complete. Cortex layers distribution of cells are normal. Nerve cells have a variety of morphology. And nerve cells arranged neatly in the hippocampus. After1day and3day in the model group,cortical parietal area of nerve cells loss a lot. Nerve cells and astrocytes have obviously changes in pathology. After giving Tong xin luo,brain tissue in general form can be intuitive to medicine group was observed on infarcts area compared with the model group decreased obviously. Observed under high power light microscopy, drug groups of neurons and astrocytes reduced damage degree, and there are new blood vessels, neurons to form.5. By ELISA method for detection of rats after acute cerebral infarction group and risk of side (left) brain serum in homogenate glial fibers acidic protein (GFAP), vascular endothelial growth factor (VEGF), angiogenin (Ang-1), endodermis chalone (ES) content changes, result:1) GFAP levels in erum:delivery after1day, model group increased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Delivery after3day, model group increased compared with sham group, has the extremely significant difference (P <0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Content of GFAP in brain homogenate:delivery after1day, model group increased compared with sham group, has the extremely significant difference (P <0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Delivery after3day, model group increased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001).2) VEGF levels in serum:delivery after1day, model group increased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Delivery after3day, model group increased compared with sham group, has the extremely significant difference (P <0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Content of VEGF in brain homogenate:delivery after1day, model group increased compared with sham group, has the extremely significant difference (P <0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Delivery after3day, model group increased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001).3) Ang-1levels in serum:delivery after1day, model group increased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Delivery after3day, model group increased compared with sham group, has the extremely significant difference (P <0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Content of Ang-1in brain homogenate:delivery after1day, model group increased compared with sham group, has the extremely significant difference (P <0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001). Delivery after3day, model group increased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001).4) ES levels in serum:delivery after1day, model group decreased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group increased, with extremely significant difference (P<0.001). Delivery after3day, model group decreased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group increased, with extremely significant difference (P<0.001). Content of ES in brain homogenate:delivery after1day, model group decreased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group increased, with significant difference (P<0.01). Delivery after3day, model group decreased compared with sham group, has the obvious difference (P<0.05). Tong xinluo group compared with model group increased, with extremely significant difference (P<0.001).6. By immune histochemical method analyse the expression of GFAP, VEGF, Ang-1, ES in rats brain parietal cortex after acute cerebral infarction group,result:1)GFAP:delivery after1day, model group compared with sham group, had a tendency to rise, has the extremely significant difference (P<0.001).Tong xinluo group compared with model group decreased, with significant difference (P<0.01). Delivery after3day, model group increased compared with sham group, has the extremely significant difference (P<0.001). Tong xinluo group compared with model group decreased, with extremely significant difference (P<0.001).2) VEGF:delivery after1day, model group increased compared with sham group, has the obvious difference (P<0.05). Tong xinluo group compared with model group decreased, but no statistical difference. Delivery after3day, model group compared with sham group, had a tendency to rise, but no statistical difference. Tong xinluo group compared with model group decreased, has the obvious difference (P<0.05).3)Ang-1:delivery after1day, model group increased compared with sham group, but no statistical difference. Tong xinluo group compared with model group decreased, with obvious difference (P<0.05). Delivery after3day, model group decreased compared with sham group, has the obvious difference (P<0.05). Tong xinluo group compared with model group increased, with extremely significant difference (P<0.001).4)ES:Delivery after3day, model group increased compared with sham group, has the obvious difference (P<0.05).Conclusion1. There is significant damage on rats’neurons after acute cerebral infarction, Tong xinluo has a certain promotion effect on recovery of neuroal function;2. Tong xinluo improve microvascular structure in rats with acute cerebral infarction, to achieve neuro protective effect;3. After acute cerebral infarction on rats,Tong xinluo probably adjust microvascular regeneration in promoting and inhibiting factors in the environment, to improve the establishment of collateral circulation in ischemia half dark area.