| Summary: | 博士 === 中山醫學大學 === 醫學分子毒理學研究所 === 98 === This study focused on endogenous and exogenous stem cells transplanted in the neural stem cells for ischemic stroke of neuroprotective properties associated with the treatment mechanism. In recent years, stem cell research has become the nerve repair and nerve regeneration of the popular subject of studies have shown that stem cells have the potential to divide, in harm organs, can be given appropriate support and repair, and even division, and replaced by renewable sources. As a result, this study using rats in the brain artery ischemic stroke reperfusion animal model to explore the endogenous stem cells in stroke, the source of their case distribution and cycle time for self-repair and neural contributions to the protection, and then from the use of before the fetal rat brain by the isolated neural stem cells to give cell treatment, evaluation of such treatment after a stroke in nerve protection and restoration of acts of effective treatment and may participate in an in-depth understanding of the elements is the means through which the elements of the mechanism.
In this study, first of all rats in the brain artery embolization surgical sutures and then reperfusion (ischemia - reperfusion) experiment, hoping to simulate a stroke after thrombolytic therapy in patients after brain tissue changes; ischemic stroke after an hour later, though immediately the implementation of blood reperfusion, but rats have caused irreversible brain of ischemic injury after nerve defect classification determined that 2 to 3 defects in rats for experimental analysis of the wind objects; by TTC staining was informed analysis of the stroke and brain injury area Brain shrinkage ratio of brain damage after a stroke in the area of 1 to 3 days up to the maximum, accounting for about 25% of the whole area of the brain; 7 days after the stroke in the brain begins to shrink, accounting for about half of the 5% of the brain, a stroke after 14 to 28 days , Brain atrophy rate stability, has accounted for about half the brain of 10%. The current study will be regular stem cells Nestin as one of the standard elements, so this experiment as a preliminary to explore the elements of endogenous stem cells of the subject. The use of immune to stain the way for a stroke after the protein in rat brain tissue for analysis and found that nestin was obviously a large number of induced stroke in 3 days after the stroke injury agency (cortex) and stroke, peripheral damage (striatum), the results of the stroke to sustainable After 14 days, 28 days after the stroke in the left cerebral cortex has a surplus of performance, type of stem cells will be displayed by the stroke was induced, but its source and distribution, is not yet clear. So the use of immune staining nestin cell in the brain tissue of stroke.
Slice in the distribution, nestin is found in a large number of strokes after three days in the neighboring ventricle, striatum and the cerebral cortex were injured in the region in seven days after the stroke, a large number of nestin cell to form a clear nestin boundary. In the range of stroke damage did not side with the injured region, the distribution of its movement from the cortex through the striatum, has been extended to the bottom of the particle layer; but after a stroke in the first 14 days, there nestin Signals in both the scope and restricted to only the ventricle, the striatum and the surrounding olfactory tubercle, in a stroke 28 days after immunization with the results of the stain to the same, only a stroke left side of the cerebral cortex were injured in the region have shown; literature that stem cells themselves have more Strong proliferative properties, the use of BrdU demarcation of the observation post-stroke newborn rat brain cells, showed that only 3 days after the stroke of the large side stroke found that the cells significantly, the majority present in the cortex, striatum and olfactory tubercle region, and in 7 days after the stroke of the new cells significantly less, showing not nestin cells by cells from newborn. The stroke group to assess the conduct, whether running or forelimb grasp, in 7 to 14 days there was a slight recovery and enhance the value, but 28 days after the show are 3 to 4 percent of the recession, show self-rescue of endogenous stem cells for stroke recovery is limited, even in the 3-day-old stroke after stroke side of a large number of nestin signal, so that these signals are not true stem cell signal, then choose another stem cell demarcation of the molecular PSA-NCAM (Polysialic acid-neural cell adhesion molecule), immuno-blot analysis of the way, the results of its performance compared to the amount of nestin expression as expected a lot less of what a truly endogenous stem cells to restore the effect is less obvious.
In addition, the study of the use of exogenous stem cell treatment of stroke in rats to observe the extent of brain damage after a stroke and to assess their conduct and to restore the situation. Exogenous stem cells from fetal rat brain before the separation, the serum in vitro cultivation of self-differentiation into neurons can be divided into experimental and stellate cells in plastic. After 1 hour rats embolism reperfusion after three hours will be derived stem cells outside the demarcation of the CM-DiI and groin by intravenous injection of stroke rats, after a cycle of migration in the body of the brain regions damaged in the calibration of up to 7 days The maximum stem cells, and the apparent stroke on the side of the striatum, the use of TTC staining confirmed that both the area of the brain injury or shrinking proportion of the treatment group than in both stroke damage in small groups; forelimb grip behavioral assessment showed that after stem cell treatment After the stroke in rats 7 to 14 days to grasp the power is better than the untreated group (P <0.01), with statistical significance, but in a stroke within 28 days after the treatment group grasp its power is still about a recession as two shows that foreign although stem cell therapy to a certain degree of protection, but do not represent in a long time, to protect the brain from stroke damage the function of thinking can be adjusted in the future as stem cell therapy in a row to give the course of treatment. And give in-depth study stem cell treatment of stroke may improve the molecular mechanism and found that inflammation-induced COX-2 and IL-1β will be given stem cell treatment, the performance of its volume will be decreased, and the protection of the protein HSP27 (heat shock protein 27 ) Will be treated for three days after the stroke was induced obvious, can protect nerve cells, able to bear the increase in apoptosis injury. Therefore, this experimental study has established that endogenous and exogenous stem cells for the treatment of stroke and the ability to Molecular changes in the future for the molecular mechanism of stroke and treatment methods can provide a more complete set of applications.
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