静松灵对体外大鼠脑神经细胞的麻醉影响研究
发布时间：2018-11-28 19:38
【摘要】：本实验以离体培养神经元细胞为研究模型,模拟麻醉过程中大脑皮层神经元所处环境,通过对神经递质及信号转导通路相关指标的测定,探究静松灵麻醉剂对中枢神经系统的麻醉效应机理,为进一步分析静松灵麻醉剂的作用机制,更合理地指导临床麻醉用药,提供理论与数据支持。 实验结果显示：原代培养大鼠脑神经细胞技术成熟、完善,可以作为麻醉机理研究的新型实验模型；LC-MS/MS检测神经递质方法可靠、准确性好、灵敏度高、重现性好。可以将该方法应用于静松灵麻醉剂的作用机制研究。 不同浓度的静松灵麻醉剂对体外培养的神经细胞分泌儿茶酚胺类神经递质有明显的抑制作用,对5-羟色胺类神经递质含量有明显的升高作用；可以有效的降低兴奋型神经递质Glu和Asp的含量,使抑制型神经递质GABA的含量升高,Gly含量先下降再升高。在对信号转导通路的实验结果中,静松灵麻醉剂对Na+-K+-ATP酶与ca2+-Mg2+-ATP酶活性产生明显的抑制作用；对cAMP信号转导通路产生了一定的抑制作用,使得cAMP的含量随时间的变化呈先上升再下降的趋势；在对NO.cGMP信号通路的影响研究中,实验结果显示,在不同浓度的静松灵麻醉剂作用下,NO的含量、NOS的活性和cGMP浓度均呈明显下降的趋势。 从而得出结论：DA和NE可能是静松灵麻醉剂产生麻醉作用的靶对象,静松灵麻醉剂对5-羟色胺类神经递质的作用机制可能是通过5-HT1A受体和5-HT3受体来发挥的；Glu与Asp在麻醉机制中起到重要作用,GABA能够参与静松灵麻醉机制,Gly在静松灵麻醉剂的麻醉机制中确切作用原理,有待进一步研究；Na+-K+-ATP酶和ca2+-Mg2+-ATP酶可能是静松灵麻醉剂产生麻醉作用的靶位之一,cAMP信号转导通路参与了静松灵的麻醉作用机制,静松灵麻醉剂对No-cGMP信号通路的抑制,可能是通过抑制a2-肾上腺素能受体所介导通路的活化来实现的。
[Abstract]:In this study, in vitro cultured neuronal cells were used as the model to simulate the environment of cerebral cortical neurons during anaesthesia, and the related indexes of neurotransmitters and signal transduction pathways were measured. To explore the mechanism of the anesthetic effect of Jingsongling anesthetic on central nervous system, and to provide theoretical and data support for further analysis of the mechanism of Jingsongling anesthetic and more reasonable guidance for clinical anaesthesia. The results showed that the technique of primary cultured rat brain nerve cells was mature and perfect and could be used as a new experimental model for the study of anesthetic mechanism, and the method for detecting neurotransmitters by LC-MS/MS was reliable, accurate, sensitive and reproducible. This method can be applied to study the action mechanism of Jingsongling anesthetic. Different concentrations of jingsongling anesthetic can inhibit the secretion of catecholamine neurotransmitters and increase the content of 5-hydroxytryptamine neurotransmitters. It can effectively reduce the content of Glu and Asp, increase the content of GABA, decrease the content of Gly and then increase the content of Gly. In the experimental results of signal transduction pathway, the activity of Na-K-ATP enzyme and ca2-Mg2-ATP enzyme was significantly inhibited by jingsongling anesthetic. The cAMP signal transduction pathway was inhibited and the content of cAMP increased first and then decreased with time. In the study of NO.cGMP signaling pathway, the experimental results showed that the content of NO, the activity of NOS and the concentration of cGMP decreased significantly under the action of different concentrations of Jingzongling anesthetic. It is concluded that DA and NE may be the target targets for the anesthetic effect of Jingzongling anesthetic, and the mechanism of the action of JZL on serotonin neurotransmitters may be played by 5-HT1A receptor and 5-HT3 receptor. Glu and Asp play an important role in the mechanism of anesthesia, GABA can participate in the mechanism of Jisongling anesthetic, and the exact mechanism of Gly in the anaesthesia mechanism of JZL needs further study. Na-K-ATP enzyme and ca2-Mg2-ATP enzyme may be one of the targets for the anaesthesia effect of jingsongling anesthetic. CAMP signal transduction pathway is involved in the mechanism of jingsongling's anaesthesia and the inhibition of No-cGMP signal pathway by jingsongling anesthetic This may be achieved by inhibiting the activation of a-2-adrenergic receptor-mediated pathways.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：S859.791
本文编号：2364063
[Abstract]:In this study, in vitro cultured neuronal cells were used as the model to simulate the environment of cerebral cortical neurons during anaesthesia, and the related indexes of neurotransmitters and signal transduction pathways were measured. To explore the mechanism of the anesthetic effect of Jingsongling anesthetic on central nervous system, and to provide theoretical and data support for further analysis of the mechanism of Jingsongling anesthetic and more reasonable guidance for clinical anaesthesia. The results showed that the technique of primary cultured rat brain nerve cells was mature and perfect and could be used as a new experimental model for the study of anesthetic mechanism, and the method for detecting neurotransmitters by LC-MS/MS was reliable, accurate, sensitive and reproducible. This method can be applied to study the action mechanism of Jingsongling anesthetic. Different concentrations of jingsongling anesthetic can inhibit the secretion of catecholamine neurotransmitters and increase the content of 5-hydroxytryptamine neurotransmitters. It can effectively reduce the content of Glu and Asp, increase the content of GABA, decrease the content of Gly and then increase the content of Gly. In the experimental results of signal transduction pathway, the activity of Na-K-ATP enzyme and ca2-Mg2-ATP enzyme was significantly inhibited by jingsongling anesthetic. The cAMP signal transduction pathway was inhibited and the content of cAMP increased first and then decreased with time. In the study of NO.cGMP signaling pathway, the experimental results showed that the content of NO, the activity of NOS and the concentration of cGMP decreased significantly under the action of different concentrations of Jingzongling anesthetic. It is concluded that DA and NE may be the target targets for the anesthetic effect of Jingzongling anesthetic, and the mechanism of the action of JZL on serotonin neurotransmitters may be played by 5-HT1A receptor and 5-HT3 receptor. Glu and Asp play an important role in the mechanism of anesthesia, GABA can participate in the mechanism of Jisongling anesthetic, and the exact mechanism of Gly in the anaesthesia mechanism of JZL needs further study. Na-K-ATP enzyme and ca2-Mg2-ATP enzyme may be one of the targets for the anaesthesia effect of jingsongling anesthetic. CAMP signal transduction pathway is involved in the mechanism of jingsongling's anaesthesia and the inhibition of No-cGMP signal pathway by jingsongling anesthetic This may be achieved by inhibiting the activation of a-2-adrenergic receptor-mediated pathways.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：S859.791
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