神经钙调蛋白在促炎细胞因子诱导神经元凋亡中作用机制

发布时间：2018-01-16 04:18

本文关键词：神经钙调蛋白在促炎细胞因子诱导神经元凋亡中作用机制　出处：《吉林大学》2005年博士论文　论文类型：学位论文

【摘要】：本实验主要利用炎症细胞因子IL-1β和TNFα作用于原代培养的皮层神经元和星形胶质细胞,探讨神经钙调蛋白是否参与了炎症因子引起的神经细胞的损伤及可能的机制。方法:用IL-1β和TNFα诱导培养大鼠皮层神经元和星形胶质细胞的损伤,采用四唑盐比色法(MTT),乳酸脱氢酶释放率(LDH)及免疫荧光方法观察细胞生存力和损伤程度, Western blot 分析检测p-JNK,p-ERK,NF-κB 的表达。结果:在皮层神经细胞中,IL-1β和TNFα与正常对照组比较,均可明显引起皮层神经元的损伤(p0.05)。而IL-1β与CsA 联合应用组,皮层神经元的生存力则明显提高,TNFα与CsA 联合应用组,皮层神经元的生存力改变不明显。在LDH 释放率测定试验中,也观察到了类似的结果。CsA 可明显降低IL-1β引起的LDH 释放率的增加。IL-1β与CsA 联合作用组,与IL-1β单独作用组比较可明显降低LDH 的释放率(p0.001),而TNFα与CsA 联合作用组,与TNFα单独作用组比较LDH 的释放率没有明显改变(p0.05)。在星形胶质细胞中炎性细胞因子IL-1β,TNFα均可引起细胞释放LDH 增多,IL-1β与CsA 可明显降低IL-1β引起的星形胶质细胞生存力降低和LDH 释放率的增加。IL-1β与CsA 联合作用组,与IL-1β单独作用组比较可明显提高细胞的生存力和降低LDH 的释放率(p0.001),而且TNFα与CsA 联合作用组,与TNFα单独作用组比较LDH 的释放率可明显降低(p0.05)。通过免疫荧光观察发现,皮质神经元中IL-1β和TNFα引起神经元凋亡数(green)和死亡数(red)的细胞数均明显增多,与正常对照组比较有显著差异(p0.05)。而CsA 组神经元凋亡及坏死数量均比IL-1β和TNFα损伤组少。CsA 与IL-1β联合应用组,神经元凋亡和坏死率与单纯
[Abstract]:In this study, the inflammatory cytokines IL-1 尾 and TNF 伪 were used in primary cultured cortical neurons and astrocytes. To investigate whether calmodulin is involved in the injury of nerve cells induced by inflammatory factors and its possible mechanism. Methods: IL-1 尾 and TNF 伪 were used to induce the injury of cortical neurons and astrocytes in cultured rats. Cell viability and damage were observed by MTT assay, lactate dehydrogenase release rate (LDH) and immunofluorescence assay. The expression of NF- 魏 B was detected by Western blot analysis. Results: in cortical neurons. Compared with the normal control group, IL-1 尾 and TNF 伪 could significantly induce the cortical neuron injury (p0.05), while the IL-1 尾 and CsA combined treatment group. The viability of cortical neurons was significantly increased in the combination of TNF- 伪 and CsA group. The viability of cortical neurons did not change significantly. In the test of LDH release rate, there was no significant change in the viability of cortical neurons. Similar results were observed. CSA could significantly reduce the increase of LDH release rate induced by IL-1 尾. IL-1 尾 combined with CsA group. Compared with IL-1 尾 alone, the release rate of LDH was significantly decreased in TNF 伪 and CsA group. Compared with TNF 伪 alone, the release rate of LDH did not change significantly. The inflammatory cytokine IL-1 尾 was found in astrocytes. TNF 伪 could induce the increase of LDH release. IL-1 尾 and CsA significantly decreased the viability of astrocytes induced by IL-1 尾 and the increase of LDH release. Compared with the IL-1 尾 alone group, the viability of the cells and the release rate of LDH were significantly increased and the release rate of LDH was decreased. Moreover, the combination of TNF 伪 and CsA group. Compared with TNF 伪 alone, the release rate of LDH was significantly lower than that of TNF 伪 alone. In cortical neurons, the number of IL-1 尾 and TNF 伪 induced neuronal apoptosis and the number of dead neurons increased significantly. The number of apoptosis and necrosis of neurons in CsA group was less than that in IL-1 尾 and TNF 伪 injury group. CSA combined with IL-1 尾 group was significantly different from normal control group. Neuron apoptosis and necrosis rate and simple
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R363

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