骨钙素对小鼠星型胶质细胞周期调控作用的初步研究

发布时间：2018-11-15 08:45

【摘要】：研究背景:抑郁症(depression),一种以情绪低落,兴趣减退为主要症状的精神科常见临床疾病,给患者带来了极大的心理痛苦,也给社会带来了沉重的经济负担。根据最新流行病学调查数据显示,抑郁症的发病率呈现出逐年上升的趋势。目前学者对于抑郁症的发病机制存在多种学说,但尚无统一定论,因此对抑郁症的发病机制研究显得尤为迫切。行使大脑功能的基本单元是神经元,然而近来研究表明星型胶质细胞(astrocyte)不仅限于对神经元的支持营养作用,自身也存在信号传递的功能。星型胶质细胞的数量和功能与神经精神疾病存在密不可分的关系。大量的科学研究表明星型胶质细胞与抑郁症等多种认知功能障碍疾病具有相关性。骨钙素(osteocalcin)作为一种成骨细胞释放的内源性多肽,具有多种生物学功能,新近研究表明骨钙素对神经系统具有潜在的生物学效应,在神经精神疾病领域发挥着一定的作用。本实验通过研究不同浓度的骨钙素对星型胶质细胞的周期调控作用,初步揭示骨钙素参与神经精神疾病的机制。目的:本实验通过研究不同浓度的骨钙素对星型胶质细胞的周期蛋白调控作用及增殖能力的影响,初步揭示骨钙素是如何参与调节星型胶质细胞的周期调控机制。方法:使用Hanks细胞培养方法进行c57乳鼠星型胶质细胞的原代培养,并使用传代培养法,传三代获得纯化的星型胶质细胞。使用cck-8试剂盒进行细胞增殖实验,实验分为三组,与450nm波长,分别测得OD值,绘制细胞增殖曲线。使用流式细胞仪Annexin V-FITC双染法流式细胞术(FCM)对细胞周期分布进行检测[6]。Western Blot凝胶电泳免疫印迹蛋白半定量方法测定周期相关蛋白CyclinD2、CyclinD3、Cyclin E、CyclinB1的相对表达量,并使用Quatity One进行条带分析。结果:30ng/ml的骨钙素作用星型胶质细胞,明显的观察到增殖效应,与对照组比较,P㩳0.05。30ng/ml的骨钙素作用星型胶质细胞,具有影响细胞周期分布的效应,且具有剂量依赖性与对照组比较,P㩳0.05。不同浓度的骨钙素影响星型胶质细胞周期蛋白的表达,如周期蛋白Cyclin D2、D3、E、B1均有不同程度的表达量改变与对照组比较,P㩳0.05。
[Abstract]:Background: depression (depression), a common psychiatric clinical disease characterized by depression and decline in interest, has brought great psychological pain to patients and a heavy economic burden to the society. According to the latest epidemiological survey data, the incidence of depression has been increasing year by year. At present, there are many theories about the pathogenesis of depression, but there is no unified conclusion, so it is urgent to study the pathogenesis of depression. The basic unit of brain function is neurons. However, recent studies have shown that astrocyte (astrocyte) is not only limited to supporting the nutritional function of neurons, but also has the function of signal transduction. The number and function of astrocytes are closely related to neuropsychiatric diseases. A large number of scientific studies have shown that astrocytes are associated with many cognitive disorders such as depression. Osteocalcin (osteocalcin), as an endogenous peptide released by osteoblasts, has many biological functions. Recent studies have shown that osteocalcin has potential biological effects on nervous system and plays a certain role in neuropsychiatric diseases. In this study, we studied the effects of different concentrations of osteocalcin on the cycle of astrocytes, and revealed the mechanism of osteocalcin involved in neuropsychiatric diseases. Aim: to investigate the effects of different concentrations of osteocalcin on the cyclin regulation and proliferation of astrocytes, and to explore how osteocalcin is involved in the regulation of astrocyte cycle. Methods: the primary culture of c57 rat astrocytes was carried out by Hanks cell culture method, and the purified astrocytes were obtained by the method of subculture. The cck-8 kit was used to carry out the cell proliferation experiment. The experiment was divided into three groups. The OD value was measured with 450nm wavelength and the cell proliferation curve was plotted. The cell cycle distribution was detected by flow cytometry (FCM) with flow cytometry (Annexin V-FITC) double staining method. [6]. Western Blot gel electrophoresis was used to detect the relative expression of cyclin B1 by Western blot. Quatity One was used to analyze the bands. Results: compared with the control group, the effect of osteocalcin of 30ng/ml on astrocyte proliferation was obvious. Compared with the control group, osteocalcin of P?0.05.30ng/ml had an effect on the distribution of cell cycle. In a dose-dependent manner, compared with the control group, P0. 05. Different concentrations of osteocalcin affected the expression of cyclin in astrocytes. For example, the expression of cyclin Cyclin D _ 2, D _ 3, E _ (1) was different from that of the control group (P < 0.05).
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.4

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