| Objective Ischemia cerebral vascular disease has become one of frequently occurring illness and common diseases to mid-aged people with higher incidenced disease, higher death and mutilation rate. With the profound investigation into cerebral arterial thrombosis, it could induce the injury of ischemia-reperfusion after bloodstream reviving recanalization. The therapeutic angiogenesis established a new channel to angiogenesis of ischemia cerebral vascular disease. Therefor, it is necessary to properly control the process of therapeutic angiogenesis which is effective to improve cerebral ischemia. Vascular endothelial growth factor is a kind of cell factor to promote angiogenesis best as we know. The activation of VEGF-VEGFR system is to cause hyperplasia endothelialis and to induce neovascularization. The new vessels, extensing from normal brain tissue to ischemic region, augment reperfusion and ventilation of brain tissue to be involved in, and lessen ischemical reperfusion injury. Acupuncture possesses multilevel, multichannel, multitarget protection and intervention role. Used the MCAO model, this research approached the mechanism of early acupuncture treating resisting cerebral ischemical reperfusion injury and based on the signal transduction of angiogenesis mediated by VEGF in brain.Methods SD rats were randomly divided into the normal control group, sham operation group, model group(ischemia 0.5h reperfusion 1 day, 2days, 4days, 8days), electroacupuncture treatment group(ischemia 0.5h reperfusion 1 day, 2days, 4days, 8days). Used the suture in MCA to establish MCAO ischemia-reperfusing model. The remarks of nerve function deficit referring to Lundy remarking standards. The acupoints of BAIHUI, SHUIGOU and ZUSANLI were selected, stimulated with slow-fast wave for 30 minutes. Observed ischemia brain tissue microstructure, detected MVD in right ipsilateral cortex after ischemia-reperfusion and electroacupuncture by immunohistochemistry SABC (spreptavidin-biotin-peroxidase complex) method. Measured the expression of VEGF mRNA of the right ipsilateral cortex by RT-PCR method. Detected by Westernblot method to investigate the content of p-Akt and PTEN in right ipsilateral cortex and used SABC method to investigate the expression of cyclinD1, caspase9 in rats' right ipsilateral cortex in the ischemic brain tissue after ischemia-reperfusion and electroacupunctureo All the datum were processed by SPSS11.0 statistics software package and all results were demonstrated by mean±standard deviation.Results (1)The electroacupuncture could significantly improve the nerve missing sign of the rats with cerebral ischemic reperfusion (p＜0.05, p＜0.01), improve the pathologic of ischemic brain tissue, and increase the microvessel density in rats' ischemic ipsilateral cortex (p＜0.05). (2)The expression of VEGF mRNA in ischemic ipsilateral cortex of model group rats were significantly higher than those of normal and sham operation group rats (p＜0.05, p＜0.01), the expression peak of VEGF mRNA was appeared at 2d; while the expression of VEGF mRNA in ischemic ipsilateral cortex of electro-acupuncture group rats were significantly higher than those of corresponding phase rats of model group(p＜0.01). (3)The content of p-Akt in ischemic ipsilateral cortex of model group rats were significantly higher than those of normal and sham operation group rats (p＜0.05, p＜0.01), the content peak of p-Akt was appeared at 1d; the content of PTEN were a little higher than those of normal and sham operation group rats, the content peak of PTEN was appeared at 2d. The content of p-Akt in ischemic ipsilateral cortex of electro-acupuncture group rats of 1d, 2d, 4d were significantly higher than those of corresponding phase rats of model group(p＜0.05), while the content of p-Akt in ischemic ipsilateral cortex of electro-acupuncture group rats of 8d was lower than those of corresponding phase rats of model group; the content of PTEN in ischemic ipsilateral cortex of electro-acupuncture group rats were little higher than those of corresponding phase rats of model group. (4)The expression of cyclinD1 and caspase9 increased in rats' ischemic ipsilateral cortex after cerebral ischemia reperfusion (p＜0.05, p＜0.01), the expression peak of cyclinD1 was appeared at 4d and that of caspase9 was at 2d. The expression of cyclinD1 increased in rats' ischemic ipsilateral cortex after electroacupuncture treatment at 1d, 2d (p＜0.01), while the expression of cyclinD1 decreased at 8d; The expression of caspase9 were lower after electroacupuncture treatment(p＜0.01).Conclusion VEGF,Akt,PTEN,cyclinD1,caspase9 participate in angiogenesis after cerebral ischemic-reperfusion injury. Electroacupuncture adjust the signal transduction of angiogenesis meditated by VEGF after cerebral ischerr.tic-reperfusion injury. This effect of Electroacupuncture adjusting could be carried out by the following ways: effectively regulating signal transduction of PI3K/Akt/PTEN, effectively regulating the activity of Akt and control the regulation of PTEN to Akt, increasing the expression of VEGF mRNA, activating the protein expression of cyclinD1 which is subsequently to regulate generation cycle in the signal pathway, restraining the protein expression of caspase9 which is subsequently pro-apoptosis gene in the signal pathway. Electroacupuncture can adjust the signal transduction system with polylink and polytarget, so as to urge angiogenesis in ischemic brain tissue and reduce pathologic process in ischemic brain tissue and improve the prognosis of CVD. |
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