Effect Of Supplementing Qi And Activating Blood Therapy And Tonifying The Kidney To Promote Generation Of The Brains Therapy On Proliferation And Differentiation Of Neural Stem Cells Cultured In Vitro And In Cerebral Ischemic Rats

ObjectiveThis dissertation is intended to do a preliminary and exploring research on the proliferation and differentiation of neural stem cells (NSC) by means of Traditional Chinese Medicine (TCM) therapies on treating ischemic stroke. We try to investigate whether different TCM therapies can promote the proliferation and differentiation of NSC and the expression of neurotrophins such as brain-derived neurotrophic factor(BDNF) and basic fibroblast growth factor(bFGF) after cerebral ischemia-reperfusion injury. Based on TCM theory and doctrine, focusing on the proliferation and differentiation of NSC, we try to explore the effects of supplementing qi and activating blood therapy and tonifying the kidney to promote generation of the brains therapy, probe into the mechanism, and establish a scientific foundation for further application.Methods1 Cerebral cortex and subcortex tissues were dissected mechanically from Wistar E14.0 rat embryos. Cell clones were acquired by using serum-free primary and passage culture. Limiting dilution was used to obtain sub cell line clone from the same cells. The clone cells were identified with Nestin and bromodeoxyuridine(BrdU) specific monoclonal antibody by immunofluorescence staining, so the ability of self-renewal and proliferation of NSC was confirmed. The differentiated cells were identified with mature neural cells specific monoclonal or polyclonal antibody (neuron-specific enolase, glial fibrillary acidic protein, galactocerebroside)by immunofluorescence staining , so the potential ability of multipotent differentiation of NSC was confirmed.2 Rat embryonic neural stem cells were cultured and induced to differentiate by Naoluoxintong serum supplementation and Zuoguiwan serum supplementation respectively. Their proliferative changes were detected via phase contrast microscopy and specific markers of neurocytes (neuron, astrocyte and oligodendrocyte) were detected via immunofluorescence staining.3 Rat models of local cerebral ischemia-reperfusion were established by thread-occluding the middle cerebral artery (MCA).The rats were randomly divided into 4 groups: sham-operated group (as normal control group);model group; supplementing qi and activating blood therapy group (Naoluoxintong–treated group);tonifying the kidney to promote...