| Acupuncture of Chinese medical science had much good effect on treating cerebral ischemia. The mechanism of acupuncture has been accumulated by many studies. Acupuncture can treat cerebral ischemia by many pathways. One of them is that, acupuncture can increase blood flow, ameliorate blood circulation and promote neovascularization in brain.Endothelial progenitor cells (EPCs) have close relation with neovascularization (angiogenesis or vasculogenesis). Bone marrow derived EPCs are mobilized to peripheral blood effective, which is an important pathway of neovascularization. So that, we took EPCs as study objects and explored the mechanism of acupuncture treating cerebral ischemia in neovascularization initially. The results would be a basis on the thesis of acupuncture promoting neovascularization after cerebral ischemia. The main contents of research and conclusions are as follows:â‘ Isolation,culture and identification of EPCs in vitro: Mononuclear cells were isolated from rat bone marrow and induced with VEGF and bFGF. For about 7 days, the cells observed the clone-like morphology. EPCs were identified by test of immunofluorescence staining for the expression of PECAM-1/CD31, VE-cadherin/CD144, fluorescein Ulex Europaeus agglutinin-1 (FITC-UEA-1) and uptake of Dil complexed acetylated low-density lipoprotein (Dil-ac-LDL ). The results shown that, bone marrow derived mononuclear cells could become functional EPCs by VEGF and bFGF inducing.â‘¡Cerebral ischemia model and acupuncture treatment: Male SD rats were randomly and evenly assigned to normal group, sham-operation group, model group and electro-acupuncture (EA) group. Model group and EA group were taken the model of acute focal cerebral ischemia and reperfusion (IR) by occluding middle cerebral artery (MCAO). EA was delivered via acupuncture needles inserted into"quchi"and"zusanli". EA treatment was performed before and after the model made. Cerebral IR model was established by occlusion of the middle cerebral artery for 120min. The results shown that, the injury of MCAO model was steady and manageable. Nerve functions of cerebral IR rats which were treated by acupuncture got back faster than the ones of which did not be treated.â‘¢The detection of EPCs quantity and other factors in blood serum: Peripheral blood was obtained from abdominal aorta at 24hr, 48hr and 72hr time points after reperfusion. EPCs quantity was displayed using flow cytometry. Blood serum was assembled. VEGF, MMP-9 and bFGF level was measured using enzyme-linked immunosorbent assay (ELISA). The activity of the different isoforms of NOS was measured by spectrophotometry. The results shown that, the injury of cerebral IR could increase EPCs quantity. But acupuncture could regulate this change in a certain extent. The mobilization of acupuncture was significant in IR-48hr. Whereas the EPCs quantity of EA groups were lower than that in model group in IR-24hr and IR-72hr. Acupuncture could decrease iNOS activity and increase VEGF level (vs model group). MMP-9 level in EA groups were also different from model group with the time changing. In IR-72hr, bFGF was detected in model and EA group. Moreover,bFGF level in EA group was significantly higher than that in model group.There are EPCs which have differentiation ability in bone marrow. Acupuncture may mobilize EPCs and regulate its quantity in blood. The effect of acupuncture is likely by increasing VEGF, MMP-9, bFGF levels and influencing NOS activity in blood serum. |
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