Effects Of Chronic Stress Hippocampus Of Rats Tiaogan Therapy Research On Stem Cell Differentiation Mechanisms

ObjectiveIn recent years, increasing the pressure of life makes people long-term chronic stress, chronic psychological stress injury has become a threat to people’s physical and mental health of super risks, depression, mental illness is one of them. Chronic psychological stress leads to neural stem cell damage, which affects learning and memory-related diseases have become an important issue biology research community.Zang in TCM theory, the liver controlling dispersion, liver function through clear, regulate body air machine, so the organs and meridians of the gas, blood running unhindered, to maintain the body’s normal emotional activity. Emotion is closely related to "Liver catharsis" function, its function is normal or not, plays a very important role in human emotional and mental activities. The substance of the liver controlling dispersion is smooth air-smooth strip body, regulate the normal operation of the human essence, gas, god, blood, water, catharsis function of the liver is the basic form of the body organs functional activity summary. TCM believe that emotional disorder caused by the body blood running disorder, yin and yang imbalance is emotional factors cause basic pathogenesis of disease, and blood running disorders and psychological stress in the pathophysiology of modern theory of understanding about the disease have many similarities, two person of bad mood inducing or disease-causing internal mechanisms have many similarities. Catharsis liver failure due to liver depression caused more by the long-term poor emotional. The clinical manifestations of depression, malaise, chest tightness, inflation threat, no appetite, insomnia, paranoia consider good, sad to cry, good long sigh based disease, these symptoms are similar to depression and chronic psychological stress. Catharsis with chronic psychological stress and TCM LIVER like, especially in the emotional depression lead to liver failure catharsis, is closely related to qi stagnation. Can be considered, the liver that is catharsis liver failure is an important part of the pathogenesis of chronic stress depression. Stagnation clinically demonstrated systemic stress disorder, actually neuroendocrine immune function disorders, which may be a common pathological basis of chronic stress depression and liver failure catharsis of Stagnation. Therefore considered chronic psychological stress are Chinese cause emotional category.Studies have confirmed that liver main Shuxie Emotions functions not only have a moderating effect on the HPA axis, and have a moderating effect on the central nervous mechanism. Treatment of liver qi stagnation disease psychological stress has been proved by many studies to be an effective therapy, liver controlling dispersion can improve the chronic stress depression in rats hippocampal neural stem cell damage, improve mood activity and learning and memory. Shugan Recipe Sini differentiate into neurons by activating the Wnt signaling pathway, promoting neural stem cells in the hippocampus. The team preliminary study found that Sini can promote Wntl, high levels of (3-catenin expression and activation of canonical Wnt signaling pathway promotes the proliferation and differentiation of NSCs. Sini can significantly increase the expression of Wnt5a and JNK, activation of non-canonical Wnt signaling pathway that level on NSCs, liver controlling dispersion mainly to facilitate the migration and differentiation of NSCs. Fine Contrast origin stem cells (SCs) and the birth of Chinese medicine, the role and development of the law, many scholars from various angles found similar effects of these two substances in the body, stem cells through the perspective of the innate essence and kidney is the innate essence meaning discussed, the innate essence of physiological function of stem cells is very similar to traditional Chinese medicine, refined form and the birth of the body stem cells present in the body is similar in form. Stem cells are a class of self-renewal and multi-differentiation potential of highly proliferative cells that have the ability to divide indefinitely and can differentiate into a variety of cells, including neural stem cells. Proliferation and differentiation of chronic psychological stress on hippocampal neural stem cells, liver qi stagnation disease Treatment of psychological stress has been proved by many studies to be an effective therapy, especially to improve the stress on hippocampal damage.Category chronic psychological stress injury are Chinese cause emotional injuries, TCM Viscera theory, the liver controlling dispersion adjusting qi is important for regulating the body’s emotional activity. Therefore, the neural stem cell proliferation and differentiation mechanisms angle Study chronic psychological stress injury hippocampal Liver dispersion, from the cellular level to elucidate the Liver dispersion connotation, rich and emotional as TCM liver cause theory, discuss kidney Tibet vent essence of interoperability relationships, and provide theoretical and experimental basis for the application Tiaogan treatment method combined with stem cell technology in the clinical treatment of major ailments.MethodsTake 8-week-old SPF adult male wistar rats for the study, using chronic unpredictable mild stress (shock, pinch the tail, fasting, restraint, ban water, ice swimming, tilt, dampness) The chronic psychological stress rats. Experimental observation of the effect of water maze Shugan Recipe Sini chronic psychological stress rats weight, reflect anhedonia sugar consumption test, reflecting activity and space exploration capabilities open field test and reflect on learning and memory abilities; by electron microscopy observation of Shugan Recipe Sini San on chronic psychological stress and structural changes in the number of synapses in rat hippocampal DG area; by Westen-blot and RT-PCR method to observe Shugan Recipe Sini chronic psychological stress rats hippocampus DG area of hippocampus neural stem cells with β-catenin, JNK Rho signaling protein expression and β-catenin, Wnt3a, Wntll, Wnt5a, JNK differentiation associated Wnt signaling pathway,, Rho gene expression.Result:1.Effect of Sini Powder on Chronic Stress Induced Depression Model Behavior Science1.1. Weight:30d and 15d consistent statistical results. The control group was significantly higher than the rest of body weight gain model group (P<0.05).1.2. sugar consumption test:modeling 30d, the control group, fluoxetine group, low-dose group Sini sucrose preference is higher than the model group (P<0.05).1.3. The open field test:① modeling 15d:blank group, the total distance, average speed, the central zone residence time than the model group(P<0.05). ② modeling 30d, the control group, fluoxetine group, middle dose group movement Sini total distance, average speed, the central zone residence time, the central area away than the model group (P <0.05)1.4. The water maze test:place navigation latencies, space exploration 120s across platforms frequency was no significant difference (P<0.05).2.Effect of Sini Powder on the number of synapses in hippocampal DG area and ultrastructureUltrastructure of the DG district found the control group shows the number of rat hippocampus synaptosomes, large synaptic structure, organelles such as mitochondria clear, Golgi apparatus, endoplasmic reticulum and other structures, relatively dense postsynaptic thick, is formed between the dendrites of nerve cells, seen a lot of microtubules in dendrites, mitochondria found no degeneration. Model group compared with the control group, the dendrites nerve filaments and microtubuIes significantly dissolve significantly fewer number of synapses, synaptic structure significantly smaller, occurrence of significant vacuolar degeneration of mitochondria, the cell structure is not clear. Fluoxetine group and model group dendritic microtubules clear, and no more than the amount of synaptic mitochondria vacuoles, clear cell structure. Sini seen in low dose Photos dendrites formed between nerve cells, the number of synapses between dendritic structure formation decreased synaptic structure shorter section dendrites decreased microfilaments, stained areas appear empty some mitochondrial swelling, cristae shorter. Sini visible dose Photos dendrites formed between nerve cells, synapses between dendrites formed somewhat reduce the number of synapses, synaptic structure significantly smaller, showing mitochondria, found no mitochondrial degeneration. Dendritic Sini high dose group the nerve cell formation, the formation of synapses between dendrites, increasing the number of synapses, synaptic structure becomes large, visible mitochondria, a small amount of some mitochondrial vacuolar degeneration, edema is not obvious.3.Effect of Sini Powder on β-catenin, Wnt3a, Wnt11, Wnt5a, JNK, Rho gene expression in hippocampal DG area3.1. Wnt11:control group, fluoxetine group, Sini low dose group compared with model group increased expression (P<0.05).3.2. Wnt5a:control group, fluoxetine group, Sini low dose group compared with model group increased expression(P<0.05).3.3. Wnt3a:fluoxetine group, Sini low, medium and high dose groups compared with model group increased expression (P<0.05).3.4. JNK:the remaining five groups are the expression of the model group increased (P< 0.05).3.5. Rho:control group, fluoxetine group, Sini dose group compared with model group increased expression (P<0.05).3.6. β-catenin:fluoxetine group; Sini dose group compared with model group increased expression (P<0.05).4.Effect of Sini Powder on JNK, Rho influence, β-catenin protein expression in hippocampal DG area4.1. JNK:control group, fluoxetine group, Sini low dose group, Sini dose group compared with model group increased expression (P<0.05).4.2. Rho:control group, fluoxetine group, Sini dose group compared with model group increased expression (P<0.05).4.3. β-catenin:fluoxetine group, Sini dose group compared with model group increased expression(P<0.05).Conclusion1.Effect of Sini Powder on Chronic Stress Induced Depression Model Behavior ScienceWhether Western medicine antidepressant fluoxetine, or Chinese medicine prescription Sini liver qi stagnation can not reverse this situation because of chronic stress causes weight of slow growth; Sini low and high dose groups should improve due to chronic shock caused by the lack of pleasure in rats, decreased interest depressive symptoms; chronic stress to a certain degree of time and led to rats reduced excitability of rat activity and exploratory behavior are affected, Tiaogan medicine Sini this depression can be, but with different doses of Sini improve the depressive symptoms are not exactly the same, indicating that the liver controlling dispersion function can be improved as some symptoms of depression chronic stress appears. Chronic psychological stress on learning and memory and hippocampal damage neural stem cells with the time and extent, may be followed by the modeling method, the environment, the use of rodent species, also have a certain relationship.2.Effect of Sini Powder on the number of synapses in hippocampal DG area and ultrastructureThe results of this study may indicate that the efficacy of liver qi stagnation Sini have scattered, whether low, medium and high doses, can improve the chronic stress depression in rats hippocampal neural stem cell damage, increasing the number of synapses, synaptic ultra improve micro-structure damage, and Sini, the high-dose group, especially high-dose group more effective. In promoting the hippocampal neural stem cells differentiate into neurons aspect, Sini high-dose group was better. That the liver controlling dispersion in the proliferation and differentiation of neural stem cells, primarily manifested as promote differentiation.3.Effect of Sini Powder on β-catenin, Wnt3a, Wnt11, Wnt5a, JNK, Rho gene and protein expression in hippocampal DG areaThe different doses of Sini Powder on gene expression and protein expression, although slightly different, but not statistically significant, indicating that Sini Powder regulation of the Wnt pathway with a dose-related genes and proteins unrelated. Sini Powder can promote the differentiation of hippocampal neural stem cells, the mechanism may promote differentiation by Wnt pathway activation and regulation to promote β-catenin, Wnt3a, Wnt11, Wnt5a, JNK, Rho genes were upregulated, prompting the β-catenin, JNK, Rho regulated genes and protein expression, thereby increasing the proportion of hippocampal neural stem cells to differentiate into neurons. Liver dispersion hippocampal damage can partially reverse the chronic stress caused by drug use Shugan Sini may improve due to chronic stress caused by the decrease in hippocampal neural stem cells, promote hippocampal neural stem cells to differentiate into neurons. The regulation of the substance to the relationship between the liver and kidney Tibet vent interoperability of neural stem cell proliferation and differentiation, mainly reflected in the "vent" in terms of promoting the differentiation of neural stem cells. Innovation1. To propose new hypotheses:On the cellular level, that the theory of TCM Viscera mainly hidden in the kidney of innate essence associated with stem cells, and liver stores like the liver controlling dispersion function can cause can cause the neural stem cells differentiation, liver and kidney Tibet vent Shuxie interoperability features not only a fine reproduction of catharsis, but also the performance impact on the proliferation and differentiation of neural stem cells in, the activation of the Wnt pathway, mainly in the non-classical pathway activation promote hippocampal neural stem cells to differentiate into neurons.2. The first time wistar rats in vivo experiments, the regulation from the perspective of the proliferation and differentiation of neural stem cells to explore Liver dispersion, kidney, liver and kidney essence Tibet vent substance interoperability relations, to expand the theory of TCM viscerait provides a new way of thinking. Problems and ProspectsPrimarily by chronic stress model of depression that is liver depression model, in vitro binding preliminary team hippocampal neural stem cells have been studied in vivo studies are not performed kidney model, the drug in vitro and in vivo environment environment there are some pharmacological effect the difference, which is defective in this experiment. Sini How Wnt signaling in the regulation of neural stem cells, namely their corresponding mechanism remains unclear. Therefore, the need for further study Wnt signaling pathway, determine its functional component, Sini regulation of its components; Sini for the compound, which act as drugs, medicaments which ingredients, these active ingredients through What pathway modulator, which for purposeful intervention neurogenesis research treatment of neurological disease or injury and drug targets, in-depth innovation and basic research of traditional Chinese medicine theory, revealing its scientific connotation, to clarify its scientific conduct innovative research, has very important.