The Effects Of YiQi-HuoXue Therapy On Expressions Of TSP-1, TSP-2 And Their Receptor CD36 In Intracerebral Hemorrhagic Rat Brains

Object and Background: Intrcerebral Hemorrhage(ICH)is a common severe clinical disease, belonging to the category of stroke in Traditional Chinese Medicine(TCM). It brings heavy burden to society, with high incidence, death and mutilation rate. Since the view of Qixu Xueyu posed by a famous doctor of Qing dynasty, Wang Qingren, who initially invented the Buyang Huanwu Decoction, Yi-Qi Huo-Xue(YQHX) therapy has been widely used in clinic. The modern researches have found that YQHX therapy can improve the blood flow in the brain hemorrhage zone, rectify the abnormal blood rheology changes in brain and so on. However, the vessel system of the hematoma was damaged. Only through the reconstruction of capillary vessel system in the focal zone, can the perfusion in the hematoma zone be restituted, the microenvironment of the focus be improved and the repair of nervous system be accelerated. In recent years, most researches of reconstruction of capillary system have paid attention to VEGF. But the angiogenesis is completed by the kinetic balance of promotive and inhibitive factors. Thrombospondins-1 (TSP-1) and TSP-2 can combine its receptor CD36 and inhibit the angiogenesis. Therefore, we suppose that through the influence on the expression of TSP-1, TSP-2 and CD36, the Yiqi Huoxue therapy may adjust the inhibition of angiogenesis in injured region, improve the absorption of the hematoma and accelerate the repair of brain tissue.By observing the effects of YQHX therapy on angiogenesis and measuring the areas of hematomas, we try to explore the distribution feature of TSP-1, TSP-2 and CD36 and space-time change rules after intervention of YQHX, to study the mechanism of YQHX therapy on reconstitution of capillary vessel system in the hematoma area after ICH initially.Methods: 180 SD rats were randomly divided into six groups including normal control group, sham control group, ICH model control group, Yiqi-treated group, HuoXue-treated group and YQHX-treated group. By injecting collagenase ICH model was made. Yiqi, HuoXue and YQHX-treated group were respectively administered at Buyang Huan wu decoction, YiQi decoction and HuoXue decoction. At 1d,4d,7d,14d, 21d, 28d and 35d after operation, rats were randomly chosen to be perfused with paraformaldehyde-PBS through ascending aorta and getting the brain. Dehydrating in sugar solution, cutting slice. Angiogenesis was observed by argentation. Image pro plus 5.0 software measured the areas of hematomas. Immunohistochemistry was used to measure the expression of TSP-1, TSP-2 and CD36 in the haematoma.Results:1. angiogenesis after Intreerebral HemorrhageAt 1d, Some distended capillarys surrounding hematoma area could be seen; At 4d, it could be seen that some capillarys had pullulated in each groups; At 7d, every group of hematoma was surrounded by a great deal of blood vessel-like reticular fibers, which began to push toward the center of the hematoma, and it was obvious in YQHX-treated group; At 14d, Mesh kind structure had already overloaded the whole area of hematoma. At the edge of the mesh, the blood vessel-like structure could be seen accidentally. The capillary segment had been seen in YQHX-treated group; At 21d, the mesh kind structure was further good in order. Obvious capillary segment could be seen in every group, and there was a little more in HuoXue and YQHX-treated group; At 28-35d, The new-born capillary beside the mesh began to obliterate.2. the area of the hematoma after Intreerebral HemorrhageAt 1d, the hematoma of the HuoXue-treated group was biger than Yiqi (p＜0.01) and YQHX-treated group(p＜0.05); Yiqi-treated group had the smallest hematoma(p＜0.01). At 4d, the model group had the bigest hematoma(p＜0.01); At 7d, the model group had the bigest hematoma(p＜0.01or p＜0.05); YQHX-treated group had the smallest hematoma(p＜0.01). At 14d, 21d, 28d, 35d, comparing with other groups of the same time, the model group had the bigest hematoma(p＜0.01); YQHX-treated group had the smallest one(p＜0.01).3. the expression of TSP-1, TSP-2and CD36 after Intreerebral HemorrhageTSP-1 was mainly expressed in blood endothelial cells and its surroundings of hippocampus, thalamencephal, basal ganglia and cortex of each group. As the time passed by, the expression of the blood endothelial cells was obvious at the edge of the haematoma and its surroundings. At 4d, it was significantly increased in each group (p＜0.01) and reached to the point; YQHX-treated group's expression was higher than that in model (p＜0.01)and HuoXue-treated group(p＜0.05). From 4d to 35d, the expression of each group turned weak gradually and tended to normal level.TSP-2 was mainly expressed in blood endothelial cells and its surroundings of hippocampus, thalamencephal, basal ganglia and cortex of each group. As the time passed by, the expression of endothelial cells of the hemorrhage and its surroundings was obvious. From 1d to 28d, the expression of each group increased gradually. From 14d to 21d, the expression of YQHX-treated group was slower than others(p＜0.01or p＜0.05). At 28d, every group achieved the peak at the same time, The expression of YQHX-treated group was stronger than model group (p＜0.05).CD36 was mainly expressed in blood endothelial cells of hippocampus, thalamencephal, basal ganglia and cortex of each group. As the time passed by, the expression of hemorrhage endothelial cells was obvious. At 4d, the expression was significantly increased and reached the peak in each group, model group's expression was higher than that in HuoXue-treated group(p＜0.01). From 7d to 14d, the expression became weak gradually. From 14d to 28d the expression of each group increased gradually and reached peak. At 28d, the expression of YQHX-treated group was stronger than model group(p＜0.01).Conclusion.1. The expression of TSPs was upregulated following intracerebral hemorrhage, which of CD36 present double-peak expression.2. By adjusting the expressions of TSP-1, TSP-2 and CD36 in the rat brain of ICH, YQHX therapy may relieve the inhibition of angiogenesis, accelerate the absorption of hematoma and promote the tissue repair of brain.