Effect Of Qi-tonifying And Stasis-eliminating Therapy On Activition Of VEGFR-2 In Brain Of Intracerebral Hemorrhagic Mice

Background and prupose:Intracerebral hemorrhage (ICH) accounts for 10-30% of all strokes, with high morbidity and mortality. Prognosis is poor for present insufficient effectiveness of hematoma eliminating and supportive treatments and half death in acute stage, as well as 50% mortality within 30 days. ICH can be categorized in "stroke" of Traditional Chinese Medicine. Buyang Huangwu decotection was invented by a famous doctor Wang Qingren in Qing dynasty, which is used extensively in clinical practice, and get an identified efficient with the function of Qi-tonifying and stasis-eliminating. The recent studies on Buyang Huanwu decotection mainly focus on inibition of inflammatory reaction, apoptosis, rectification of hemorheological abnormal, reduction excitability toxicity, lessening injury of brain tissue, promotion of neurons and brain tissue. As the most potent mitogen, vascular endothelial growth factor(VEGF) must bind to its receptor-vascular endothelial growth factor receptor-2, makes it phosphorylate, then finishes a series of signal conductions downstream, plays the role of promoting proliferation, migration, survive of endothelial cells(ECs), as well as vasopremeability, in order to form a mature and steady vessel structure. In this study, ICH model was induced by injection collagenase type VII into right globus pallidus stereotaxically, and the Buyang Huanwu decoction as the typical prescription of Qi-tonifying and stasis-eliminating therapy was used to intervene in ICH mice. Changes of ICH mice in neuroethology and level of p-VEGFR-2 which is detected by Immunohistochemistry were observed. The prupose of our study is to discuss the effect that Qi-tonifying and stasis-eliminating therapy plays on p-VEGFR-2 in the brains of ICH mice, therefore try to reveal the mechanisms how the Qi-tonifying and stasis-eliminating therapy promots angiogenesis following ICH.Method:25 Kunming mice were divided into 5 groups randomly, as sham control(n=5), ICH group(n=5), ICH+SU5416 group(n=5), BYHWT group(n=5), BYHWT+SU5416 group(n=5). ICH model was induced by injection collagenase type VII into right globus pallidus stereotaxically. BYHWT group and BYHWT+SU5416 group were administered orally with Buyang Huanwu decoction, while sham control, ICH group, ICH+SU5416 group were administered orally with distilled water with an equally amount of Buyang Huanwu decoction. ICH+SU5416 group and BYHWT+SU5416 group were injected SU5416 intraperitoneally The neuroethology was observed on the first, third and seventh day following the surgery, and recorded the scores of it; Mice were executed the seventh day of surgery, decoherenced the skull and get the tissue of the brain around haematoma to detect the level of p-VEGFR-2 by immunohistochemistry. Results:1. On the first day, there is no difference amount the groups in weight, NSS, corner Turn Test, foot-fault test except sham control(P> 0.05), On the third and seventh day, the weight of ICH group is significantly heavier than ICH+SU5416 group, the weight of BYHWT group is significantly heavier than BYHWT+SU5416 group, the weight of BYHWT group is significantly heavier than ICH group (P<0.05);The NSS in ICH group is significantly lower than ICH+SU5416 group, the NSS in BYHWT group is significantly lower than BYHWT+SU5416 group, the NSS in BYHWT group is significantly lower than ICH group (P<0.05); In corner Turn Test and foot-fault test, the times for right-turn and foot-fault in ICH group are much less than ICH+SU5416 group, the times for right-turn and foot-fault in BYHWT group are much less than BYHWT+SU5416 group, the times for right-turn and foot-fault in BYHWT group are much less than ICH group (P<0.05)2. On the seventh day, there is no expression of p-VEGFR-2 in sham control. The expression of p-VEGFR-2 in ICH group and BYHWT group are significantly higher than their corresponding SU5416 groups (P< 0.05); The expression of p-VEGFR-2 in BYHWT group is significantly higher than ICH group (P<0.05); The expression of p-VEGFR-2 is located in vWF-labeled ECs.3. On the seventh day, there is no BrdU-labeled nuclei in vessels were observed in sham-control mice. BrdU-labeled nuclei and vessels in ICH group and BYHWT group are significantly much than their corresponding SU5416 groups (P<0.05). BrdU-labeled nuclei and vessels in BYHWT group are significantly much than ICH group (P< 0.05)Conclusions:1. The blockage of the phosphorylation of VEGFR-2 can inhibite angiogenesis in the brain of ICH mice, and hindrance the recovery of Neurological behavioral function.2. Qi-tonifying and stasis-eliminating therapy might promote angiogenesis and recovery of Neurological behavioral function via elevating the level of p-VEGFR-2 in the brain of ICH mice.