【摘要】：本實(shí)驗(yàn)以離體培養(yǎng)神經(jīng)元細(xì)胞為研究模型,模擬麻醉過程中大腦皮層神經(jīng)元所處環(huán)境,通過對(duì)神經(jīng)遞質(zhì)及信號(hào)轉(zhuǎn)導(dǎo)通路相關(guān)指標(biāo)的測(cè)定,探究靜松靈麻醉劑對(duì)中樞神經(jīng)系統(tǒng)的麻醉效應(yīng)機(jī)理,為進(jìn)一步分析靜松靈麻醉劑的作用機(jī)制,更合理地指導(dǎo)臨床麻醉用藥,提供理論與數(shù)據(jù)支持。 實(shí)驗(yàn)結(jié)果顯示：原代培養(yǎng)大鼠腦神經(jīng)細(xì)胞技術(shù)成熟、完善,可以作為麻醉機(jī)理研究的新型實(shí)驗(yàn)?zāi)Ｐ�；LC-MS/MS檢測(cè)神經(jīng)遞質(zhì)方法可靠、準(zhǔn)確性好、靈敏度高、重現(xiàn)性好。可以將該方法應(yīng)用于靜松靈麻醉劑的作用機(jī)制研究。 不同濃度的靜松靈麻醉劑對(duì)體外培養(yǎng)的神經(jīng)細(xì)胞分泌兒茶酚胺類神經(jīng)遞質(zhì)有明顯的抑制作用,對(duì)5-羥色胺類神經(jīng)遞質(zhì)含量有明顯的升高作用；可以有效的降低興奮型神經(jīng)遞質(zhì)Glu和Asp的含量,使抑制型神經(jīng)遞質(zhì)GABA的含量升高,Gly含量先下降再升高。在對(duì)信號(hào)轉(zhuǎn)導(dǎo)通路的實(shí)驗(yàn)結(jié)果中,靜松靈麻醉劑對(duì)Na+-K+-ATP酶與ca2+-Mg2+-ATP酶活性產(chǎn)生明顯的抑制作用；對(duì)cAMP信號(hào)轉(zhuǎn)導(dǎo)通路產(chǎn)生了一定的抑制作用,使得cAMP的含量隨時(shí)間的變化呈先上升再下降的趨勢(shì)；在對(duì)NO.cGMP信號(hào)通路的影響研究中,實(shí)驗(yàn)結(jié)果顯示,在不同濃度的靜松靈麻醉劑作用下,NO的含量、NOS的活性和cGMP濃度均呈明顯下降的趨勢(shì)。 從而得出結(jié)論：DA和NE可能是靜松靈麻醉劑產(chǎn)生麻醉作用的靶對(duì)象,靜松靈麻醉劑對(duì)5-羥色胺類神經(jīng)遞質(zhì)的作用機(jī)制可能是通過5-HT1A受體和5-HT3受體來發(fā)揮的；Glu與Asp在麻醉機(jī)制中起到重要作用,GABA能夠參與靜松靈麻醉機(jī)制,Gly在靜松靈麻醉劑的麻醉機(jī)制中確切作用原理,有待進(jìn)一步研究；Na+-K+-ATP酶和ca2+-Mg2+-ATP酶可能是靜松靈麻醉劑產(chǎn)生麻醉作用的靶位之一,cAMP信號(hào)轉(zhuǎn)導(dǎo)通路參與了靜松靈的麻醉作用機(jī)制,靜松靈麻醉劑對(duì)No-cGMP信號(hào)通路的抑制,可能是通過抑制a2-腎上腺素能受體所介導(dǎo)通路的活化來實(shí)現(xiàn)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.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：S859.791

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