The Effect Of Acupuncture On Controlling Vessel,Governing Vessel And Bladder Channel On The Neural Stem Cell Proliferation And Differentiation In The Neonatal SD Rat After Hypoxia-ischemic Insult

Background Hypoxia-ischemic encephalopathy (HIE) is one of the most severe diseases inthe neonate, it characterized by the mass degeneration and death of the neural cells in the brain which result in the serious nervous system dysfunction of sufferer. And nowadays there is no effective treatment to this frequent occurring disease in western medicine. According to the theory of traditional Chinese medicine, we adopted the principle of "supply earlier heaven essence, regulate root of later heaven, restore brain marrow, strengthen bone, resolve tetany" to use acupuncture on Controlling Vessel , Governing Vessel and Bladder Channel to treat the HIE and had achieved a sound curative effect. However, the mechanism of acupuncture-therapy on HIE has not completely defined yet. The discovery of the stem cell in the CNS for decades had raise a new hope for the therapy of the CNS disease. There are a close relationship between the potential of the proliferation and differentiation of neural stem cells and the change of the content of neural growth factors in the brain, on the other hand, many experiments had reveal that acupuncture can stimulate human or animals to secrete a lot of neural growth factors when curing the CNS disease. Combining the biological feature of the neural stem cell and the therapeutic effect of acupuncture, we assume that acupuncture can promote the proliferation and differentiation of neural stem cells through reforming the mini-circumstance in the brain of HIE sufferer (such as changing the secretion of neural growth factors) after the HIE had occurred. And the new born cells stimulated by acupuncture furthermore compensate the loss of mass neurons during HI insult and as a result contribute to the recovery of dysfunction after HIE.Purpose In our current study, we detect the new born neural cells in the HIE model animalbrain in different time point during performing acupuncture on Controlling Vessel , Governing Vessel and Bladder Channel, and furthermore analysis the possible reason of the proliferation anddifferentiation of neural stem cells, finally, reveal the cytology mechanism of the acupuncture-therapy on the HIE and even other disease.Materials and methods Postnatal day 7 Sprague-Dsawley rats underwent left carotidartery ligation followed by 8% O2 exposure for 2 hours. Animals were divided into 3 groups: Acupuncture group, control group and normal group, after establishing the HIE animal model, acupuncture was performed once daily on Controlling Vessel > Governing Vessel and Bladder Channel, which contain some well known points beneficial to cure many diseases in animals and human. Control group and normal group did not accept acupuncture. Bromodeoxyuridine (BrdU) was administered intraperitoneally twice daily to label the proliferating cells , cell phenotypes and quantities of the progenitors in the brain of each group were compared by anti-Brdu immunohistochemistry in the time point of 3d >. 7d% 14d and 28d, and double-labeling Immunofluorescence were adopted to observe the differentiation of the new born neural cells 40days after the acupuncture had performed.Results 1. The HIE model animals is hypogenetic, and remarkable neural cell degeneration or necrosis was detected in the ipsilateral cerebral cortex and hippocampus in the model animals. 2. Anti-BrdU imunohistochemistry revealed that Brdu positive cells spread around the brain of normal neonatal SD rats, and the cells density in the DG of hippocampus and the subventrical zone are especially higher than other areas. 3. There are a cluster of BrdU positive cells in the necrotic areas of the cerebral cortex and hippocampus after HI insult. A significant increase of the neural precursor cell in the cerebral cortex and hippocampus from day 3 to day 28 after hypoxia-ischemic insult compared with normal group.4 .The quantities of BrdU-positive cells in the acupuncture group is much higher than those in the control group at day 14 and day 28 after the acupuncture-therapy, however, the cells quantities has no significant difference between the acupuncture group and the control group from day 3 to day 7 after the acupuncture is performed, double-labeling Immunofluorescence reveal that the Brdu-positive cells stimulated by acupuncture 40 days later are mostly coexist with the neuron marker NSE, and only a few coexist with the astrocytes marker GFAP.Conclusion l.The development and the cerebral pathologic change of the neonate animals display that the improved approach to establish a HIE model has the advantage of low mortalityand credible pathological change, so , it can be used in our current study. 2. HI insult stimulates the stem cell proliferation in the cerebral cortex and hippocampus in the neonatal SD rat. 3.Acupuncture on Controlling Vessel % Governing Vessel and Bladder Channel promote the potential of the neural stem cell proliferation after HI insult in the neonatal rat. 4.The BrdU-positive cells are mostly differentiate into neuron ,this result suggest that the new born cells stimulated by acupuncture may be a compensation to the loss of mass neurons and furthermore contribute to the recovery of dysfunction after HIE.