【摘要】：目的：在大鼠皮層神經元缺血性損傷模型中,探討兩種半胱氨酰白三烯受體(CysLT1R和CysLT2R)拮抗劑的抗氧化作用。 方法：以CysLT1R拮抗劑孟魯司特和CysLT2R拮抗劑HAMI3379預處理,對體外培養(yǎng)大鼠皮層神經元進行缺糖缺氧／恢復(OGD/R)或H202處理,觀察神經元活化氧(ROS)產生、線粒體膜電位以及神經元存活率變化,以及兩種藥物在這些過程中的作用。同時,以RNA干擾抑制CysLT1R和CysLT2R表達,觀察以上變化。 結果：OGD處理1h后,神經元ROS產生增加,在恢復0.5h時達高峰；0.01-1μM孟魯司特和HAMI3379能輕度但顯著地抑制OGD/R誘導的ROS產生(P0.05)。OGD/R誘導神經元線粒體膜電位下降,0.01～1μM孟魯司特有輕度抑制作用(P0.05),而HAMI3379無明顯作用。外源性H202濃度和時間依賴性損傷神經元,孟魯司特和HAMI3379在0.1～1μM濃度范圍內,能輕度但顯著地減輕H202誘導的損傷(P0.05)。CysLT1R siRNA和CysLT2R shRNA對以上反應均無顯著影響。 結論：孟魯司特和HAMI3379在神經元缺血性損傷中,均有輕度抗氧化作用,且可能是不依賴受體的作用。 目的：初步研究大鼠原代小膠質細胞半胱氨酰白三烯受體表達情況。 方法：取出生24h的SD新生鼠,從皮層組織分離、純化原代小膠質細胞。通過免疫熒光染色方法,觀察原代小膠質細胞半胱氨酰白三烯受體CysLT1、CysLT2受體以及GPR17的表達情況,確定是否存在共定位；并且,使用激動劑LPS、NMLTC4誘導,觀察受體的表達變化。 結果：分離、純化并獲得原代小膠質細胞,免疫熒光染色結果顯示,CysLT1、 CysLT2受體和GPR17都有表達,它們之間存在明顯的共表達(即CysLT1R/CysLT2R、CysLT1R/GPR17和CysLT2R/GPR17)。LPS能刺激CysLT2受體發(fā)生內移,并有一定的聚集；而CysLT2受體激動劑NMLTC4誘導的CysLT2受體也誘導內移和聚集,但比LPS弱。 結論：原代小膠質細胞表達半胱氨酰白三烯受體CysLT1、CysLT2受體和GPR17,并存在明顯的共定位；炎癥誘導劑LPS能夠誘導CysLT2受體聚集和內移,CysLT2受體選擇性激動劑引起的CysLT2受體聚集和內移有待進一步確認。
[Abstract]:Aim: to investigate the antioxidation effects of two cysteinyl leukotriene receptor antagonists (CysLT1R and CysLT2R) in the rat model of cortical neuron ischemic injury. Methods: CysLT1R antagonist montelukast and CysLT2R antagonist HAMI3379 were pretreated with glucose deficiency hypoxia / recovery (OGD/R) or H2O2 to observe the production of activated oxygen (ROS) in cultured rat cortical neurons in vitro. Changes in mitochondrial membrane potential and neuronal viability and the role of both drugs in these processes. At the same time, the expression of CysLT1R and CysLT2R was inhibited by RNA interference, and the above changes were observed. Results: after treatment with OGD for 1 h, the production of ROS increased and reached the peak at 0.5 h. Montelukast and HAMI3379 at 0.01-1 渭 M inhibited ROS production induced by OGD/R (P0.05), decreased mitochondrial membrane potential induced by OGD / R and slightly inhibited montelukis specific to 0.01-1 渭 M (P0.05). HAMI3379 had no obvious effect. Exogenous H2O2 concentration and time-dependent damage to neurons, montelukast and HAMI3379 in the concentration range of 0.1 ~ 1 渭 M, slightly but significantly reduced the injury induced by H2O2 (P0.05). Both CysLT1R siRNA and CysLT2R shRNA had no significant effect on the above responses. Conclusion: both montelukast and HAMI3379 have mild antioxidant effect in neuronal ischemic injury, and may be receptor-independent. Aim: to study the expression of cysteinyl leukotriene receptor in primary microglia of rats. Methods: primary microglia were isolated from cortex of 24 h-old SD newborn mice. The expression of cysteinyl leukotriene receptor CysLT1,CysLT2 receptor and GPR17 in primary microglia was observed by immunofluorescence staining, and the co-localization was determined. In addition, the expression of cysteinyl leukotriene receptor was induced by LPS,NMLTC4. Results: primary microglia were isolated, purified and obtained. The results of immunofluorescence staining showed that both CysLT1, CysLT2 receptor and GPR17 were expressed, and there was obvious co-expression between them (that is, CysLT1R/CysLT2R,). CysLT1R/GPR17 and CysLT2R/GPR17). Lipopolysaccharide (LPS) can stimulate the inward migration and aggregation of CysLT2 receptors. CysLT2 receptors induced by NMLTC4, a CysLT2 receptor agonist, also induced inward migration and aggregation, but were weaker than those induced by LPS. Conclusion: the primary microglia express cysteinyl leukotriene receptor CysLT1,CysLT2 receptor and GPR17, and have obvious co-localization. LPS, an inflammatory inducer, can induce aggregation and inward migration of CysLT2 receptors. The aggregation and inward migration of CysLT2 receptors induced by selective agonists of CysLT2 receptors need to be further confirmed.
【學位授予單位】：浙江大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R965

【參考文獻】

相關期刊論文 前4條

1 劉鋒,朱長庚;小膠質細胞激活的分子機制[J];解剖科學進展;2003年02期

2 ;Pranlukast reduces neutrophil but not macrophage/microglial accumu-lation in brain after focal cerebral ischemia in mice[J];Acta Pharmacologica Sinica;2006年03期

3 胡欣;葛求富;張緯萍;魏爾清;;半胱氨酰白三烯受體激動劑和拮抗劑對大鼠原代皮質神經元的作用[J];浙江大學學報(醫(yī)學版);2007年02期

4 魏爾清;;半胱氨酰白三烯受體調節(jié)腦內炎癥病變的研究展望[J];浙江大學學報(醫(yī)學版);2011年02期

，

本文編號：2449835