Differentiation of human adipose-derived stem cells into neuron-like cells by Radix Angelicae Sinensis

Human adipose tissues are an ideal source of stem cells. It is important to find inducers that can safely and effectively differentiate stem cells into functional neurons for clinical use. In this study, we investigate the use of Radix Angelicae Sinensis as an inducer of neuronal differentiation. Primary human adipose-derived stem cells were obtained from adult subcutaneous fatty tissue, then pre-induced with 10% Radix Angelicae Sinensis injection for 24 hours, and incubated in serum-free Dulbecco＇s modified Eagle＇s medium/Nutrient Mixture F-12 containing 40% Radix Angelicae Sinensis to induce its differentiation into neuron-like cells. Butylated hydroxyanisole, a common in- ducer for neuronal differentiation, was used as the control. After human adipose-derived stem cells differentiated into neuron-like cells under the induction of Radix Angelicae Sinensis for 24 hours, the positive expression of neuron-specific enolase was lower than that of the butylated hydroxyani- sole-induced group, and the expression of glial fibrillary acidic protein was negative. Alter they were induced for 48 hours, the positive expression of neuron specific enolase in human adipose-derived stem cells was significantly higher than that of the butylated hydroxyanisole-induced group. Our experimental findings indicate that Radix Angelicae Sinensis can induce human adipose-derived stem cell differentiation into neuron-like cells and produce less cytotoxicity.

Synaptic and blood-brain barrier structural changes in a rat epilepsy model induced by coriaria lacton Replication experiment with animals

BACKGROUND： Structural and functional synaptic changes, as well as blood-brain barrier （BBB） changes, affect the micro-environment of nervous tissue and excitation, both of which play an important role in epilepsy. OBJECTIVE： To observe synaptic and BBB ultrastructural changes in the motor cortex of a rat epilepsy model induced by coriaria lacton, and to investigate the synaptic and BBB effects on the mechanism of epilepsy. DESIGN： A randomized controlled animal experiment. SETTING： Department of Histology and Embryology, Luzhou Medical College; and Electron Microscopy Laboratory, Luzhou Medical College. MATERIALS： Twenty healthy male Sprague Dawley rats, aged 8 weeks, were chosen for this study. The rats weighed （280 ± 50） g and were supplied by the Experimental Animal Center of Luzhou Medical College. Experimentation was performed in accordance with the ethical guidelines for the use and care of animals. The animals were randomly divided into a control group and an epilepsy group, with 10 rats in each group. METHODS： This study was performed at the Department of Histology and Embryology, and Electron Microscopy Laboratory, Luzhou Medical College between February and December 2006. According to the protocol, the epilepsy group was injected with 10 μ L/100 g coriaria lacton into the lateral ventricles to establish an epileptic model. The control group rats were not administered anything. Eight days after the model was established, all rats were anesthetized with ether. The motor cortex was removed and sectioned into ultrathin sections. Synaptic and BBB ultrastructural changes were observed by electron microscopy. MAIN OUTCOME MEASURES： （1）Structural changes of three different parts of the synapses, synaptic cleft width, postsynaptic density thickness, proportion of perforation synapses, curvature of synaptic interface, and length of active zones. （2）Capillary and BBB changes （endothelium, basement membrane, pericyte, and the astrocyte endfeet）. RESULTS： （1）Curvature of synaptic interface, length of active zones, thickness of postsynaptic density, and percentage of perforation synapses increased significantly. （2）There was significant edema in the endothelium, basement membrane, and the pericyte of the epilepsy group; the electron density of the basement membrane was reduced. CONCLUSION： （1） The coriaria lacton treatment altered synaptic ultrastructure, as well as BBB characteristics, in the epileptic rat model, and also improved synaptic transmission efficiency, as well as BBB permeability; （2）Synaptic and BBB ultrastructural changes might play an important role in the mechanism of epilepsy.

Effects of movement training on synaptic interface structure in the sensorimotor cortex and hippocampal CA3 area of the ischemic hemisphere in cerebral infarction rats

BACKGROUND： Movement is an effective way to provide sensory, movement and reflectivity afferent stimulation to the central nervous system. Movement plays an important role in functional recombination and compensation in the brain. OBJECTIVE： To observe movement training effects on texture parameters of synaptic interfaces in the sensorimotor cortex and hippocampal CA3 area of the ischemic hemisphere and on motor function in cerebral infarction rats. DESIGN, TIME AND SETTING： This neural morphology and pathology randomized controlled animal experiment was performed at the Center Laboratory, Affiliated Hospital of Luzhou Medical College, China from November 2004 to April 2005. MATERIALS： A total of 32 healthy male Wistar rats aged 8 weeks were equally and randomly assigned into model and movement training groups. METHODS： Rat models of right middle cerebral artery occlusion were established using the suture occlusion method in both groups. Rats in the movement training group underwent balance training, screen training, and rotating rod training starting on day 5 after surgery, for 40 minutes every day, 6 days per week, for 4 weeks. MAIN OUTCOME MEASURES： Texture parameters of synaptic interfaces were determined using a transmission electron microscope and image analyzer during week 5 following model induction. The following parameters were measured： synaptic cleft width; postsynaptic density thickness; synaptic interface curvature; and active zone length. Motor function was assessed using balance training, screen training, and rotating rod training. The lower score indicated a better motor function. RESULTS： The postsynaptic density thickness, synaptic interface curvature, and active zone length were significantly increased in the sensorimotor cortex and hippocampal CA3 area of the ischemic hemisphere of rats from the movement training group compared with the model group （P 〈 0.05 or 0.01）. Curved synapses and perforated synapses were seen in the sensorimotor cortex and hippocampal CA3 area at the ischemic hemisphere of rats from the movement training group, while fiat synapses were found in the model group. Balance training, screen training, and rotating rod training scores were lower in the movement training group than the model group （P 〈 0.05）. CONCLUSION： Movement training enhances synaptic plasticity in the sensorimotor cortex and, hippocampal CA3 area at the ischemic hemisphere of cerebral infarction rats, and promotes the recovery of motor function.

Altered mitochondria and Bcl-2 expression in the hippocampal CA3 region in a rat model of acute epilepsy

BACKGROUND： Previous studies have shown that the mitochondrial structure and function are damaged in animal models of epilepsy. In addition, the Bcl-2 protein is capable of regulating mitochondrial stability. OBJECTIVE： To observe and validate changes in mitochondrial structure and Bcl-2 expression, and to analyze these characteristics in the hippocampal CA3 region of rat models of epilepsy. DESIGN, TIME AND SETTING： This randomized, controlled, animal experiment was performed at the Laboratory of Electron Microscopy and Department of Histology and Embryology, Luzhou Medical College between 2007 and 2008. MATERIALS： Coriamyrtin was provided by the Pharmacy Factory of West China University of Medical Sciences. The primary and secondary antibodies were provided by Zhongshan Goldenbridge Biotechnology, Beijing. METHODS： A total of 44 adult, male, Sprague Dawley rats were randomly divided into control （n = 11） and epilepsy （n = 33） groups. Rats in the epilepsy group were induced by coriamyrtin （50 μ g/kg）, which was injected into the lateral ventricles. The rats were then observed at 3, 6, and 24 hours after epilepsy induction, with 11 rats at each time point. Epilepsy was not induced in rats from the control group. MAIN OUTCOME MEASURES： Pathological changes in the hippocampal CA3 region were observed by light microscopy; Bcl-2 expression was analyzed by immunohistochemistry; and mitochondrial changes in the hippocampus were observed under transmission electron microscopy. RESULTS： （1） The control group displayed very little Bcl-2 protein expression in the hippocampal CA3 region. However, after 3 hours of epilepsy, expression was visible. By 6 hours, expression peaked and then subsequently decreased after 24 hours, but remained higher than the control group （P 〈 0.05）. （2） Mitochondria were damaged to varying degrees in the epilepsy groups. For example, mitochondria edema, cristae space increase, and disappearance of mitochondria were apparent. Moreover, mitochondrial damage occurred prior to pathological changes in the neurons and nucleolus. CONCLUSION： Bcl-2 expression and mitochondrial damage increased in the hippocampal CA3 region in rats with epilepsy. Moreover, mitochondrial damage occurred prior to increased Bcl-2 expression and nucleolus damage.