| The anemic cerebrovascular disease which includes the temporary brain ischemia outbreak, the thrombosis and the cerebral embolism, is a common and frequently-occurring disease with high fatality rate and crippling rate. It threats humanity's health seriously, and is becoming one of the most important research subjects in the medicine domain. Anemic stroke sickness can be categorized as "winds sickness" in Chinese medicine, and its pathogenesis is "damages the brain to wind", the basic research of "collateraliasis theory" reveals that the damage of brain endothelial cell of blood vessel plays an influential role in the pathology link of the brain winds. And macrophage chemotactic factor-1β, which is produced by the brain capillaries endothelial cell, plays an important role in the micro environment of brain through paracrine or autocrine manner.Under the instruction of winds sickness theory in Chinese medcine, the brain capillaries endothelial cell was taken as a breakthrough point when we study the characteristic of the factor-1βduring its chage, which is secreted form the brain capillaries endothelial cell. The mechanism of influence by the Qingkailing inoculation fluid in the body experimental is also studied. For the sake of enriching modern biology basis of "the disintoxicating passes winds" of Qingkailing and providing the referencial methodology for the research of the traditional Chinese medicine compound prescription when it's appllied in the treatment of anemic cerebrovascular disease, the mechanism of stability maintenance of the anemic damage neuron living environment as well as its neuron protective function of the Chinese medicine,which passes through the disintoxicating winds to make the well distributed essence of food, is discussed.Objective(1) To explore the dynamic changes pattern of inflammation chemotactic factors MIP-1 beta under acute brain ischmia.(2) To explore the QKL's 24h protection mechanism of ischemia injury of rats'brain(3) To explore the QKL's 24h influence to microglia express of ischemia injury to ratsMethods(1) Useing Enzyme Linked Immunosorbent Assay (helicase determination, ELISA) to observe the dynamic changes pattern of MIP-1 beta in serum, cerebrospinal fluid (CSF) after acute ischemic damage in the process of 6h 12h,24h,72h period. We use 2,3,5-trephenyltetrazolium chloride,TTC method to observe the cerebral infarction volume changes after Permanent brain artery embolism cerebral ischemia model.(2) Useing ISA to determining the MIP-1 beta expression of PMCAO after 24h between the normal group (M group), JiaShouShu group (group S), model group (M group) and QKL treatment group (group T). Useing frozen section immunohistochemical method (IHC) to semi-quantitative analysis MIP-1 beta in four groups; Useing TTC method to observe four groups infarction volume change。(3) Using paraffin wax dyeing IHC method to observe MIP-1 beta's only receptor Chemokine CCR5 receptor 5 (C)and microglia tumor-specific markers CD11b expression after PMCAO made 24h between N group, S group, M group and T group; Using HE to observe the brain changes. Using transmission electron microscope to observe the ultrastructural changes of blood-brain barrier and the microglia organellesResult(1) After Focal cerebral infarction 6h,12h,24h,72h we tested the serum and CSF MIP-1 beta content and having found that it is higher than that of normal. Among them, the serum MIP-1 beta has reached a peak in12h point. Compared with normal there has a statistically significant (P< 0.05). CSF of MIP-1 beta has reached a peak an 6h point and continue to maintain a high level. Comparing with normal, there is a statistically significant (P< 0.05). In addition, we have found that serum MIP-1 beta content are slightly higher than which is in CSF at all four different time points. TTC dyeing results showed that model group rats'organizations came to different degree of cerebral infarction volume among 6h-72h. Among them,24h group of cerebral infarction volume significantly greater than 6h,12h,72h's group, and was statistically significant (P< 0.05, P< 0.01, P< 0.01); 72h group's infarcts are more serious than 12h group, and was statistically significant (P< 0.05).(2) After 24h Focal cerebral infarction, compared with normal group and the relative Jia Shou Shu group, the serum MIP-1 beta express in model group increased significantly (P< 0.01, P< 0.01). After QKL interventions, compared with model group the MIP-1 beta expression has increase trend, but this was not statistically significant; In the CSF models, the treatment group and normal group have been relatively increased, statistically significant (P< 0.01, P< 0.01); In brain tissue homogenate, in group compared with Jia Shou Shu groups,there has a rising trend in model group, but not statistically significant; And compared with normal, JiaShouShu model group,the QKL treatment group has a significantly increased, and was statistically significant (P< 0.01, P< 0.01).these has the uniformity with the IHC method observation. Also found in the model group and QKL treatment group, there has different degrees of brain infarcts, distributed in rats artery of the previous two/three brain areas. QKL after treatment can effectively reduce local brain infarcts volume (P< 0.01), and effectively restore embolism after the nerve dysfunction (P< 0.01).(3) 24h focal cerebral infarction, the model group's CCR5 expression are significantly increased when compared with nonnal group and JiaShouShu (P< 0.01, P< 0.01).After giving QKL injection, compared with model group,the CCR5 expression is significantly reduce in suffering side's (P< 0.01). Compared with JiaShouShu and normal group the Model group's CDllb expression is significantly increased (P< 0.01, P< 0.01).When giving QKL injection, compared with mode group,CD11b expression in suffering side are significantly reduced. (P< 0.01).Conclusion(1) Focal cerebral infarction can lead to significantly neurons impaired, microglia activation.QKL can protect neural damage.(2) in acute cerebral ischemic injury, BMEC can express MIP-1 beta and can have effect by activating microglial cell. QKL can inhibit the MIP-1 beta's combination with its receptor. It can also reduce the bad effect of activating microglia cells.through these it can maintain brain microenvironment steady. |
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