小胶质细胞不同功能状态下MAPK信号通路变化及梓醇对其影响

发布时间：2018-01-03 10:30

本文关键词：小胶质细胞不同功能状态下MAPK信号通路变化及梓醇对其影响　出处：《大连医科大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的:在前期研究工作基础上进一步观察小胶质细胞不同功能状态下MAPK(丝裂原活化蛋白激酶)信号转导通路变化及梓醇对其影响,探讨小胶质细胞功能状态变化机制及梓醇作用机制。方法:以BV2细胞株(小鼠小胶质细胞瘤)和PC12细胞株(大鼠嗜铬细胞瘤)为研究对象,分别用其来代表小胶质细胞和神经元。在不同程度缺氧及有无梓醇存在条件下培养BV2细胞,用其培养液培养PC12细胞,通过CCK-8法测定PC12细胞存活率并以此来反映BV2细胞功能,同步应用Western blot法检测BV2细胞不同功能状态下及加入梓醇前后MAPK信号转导通路JNK、p38MAPK、ERK1/2蛋白表达及磷酸化水平。结果:1.蛋白表达量应用灰度值来表示(灰度值与蛋白表达量成正比),当BV2细胞对神经元起保护作用时,p-ERK1/2表达明显高于p-p38MAPK与p-JNK,其灰度值分别为:0.277±0.010、0.221±0.019和0.163±0.003;当BV2细胞对神经元起损伤作用时,p-p38MAPK与p-JNK表达较保护作用时进一步增强,灰度值分别为:0.398±0.005和0.495±0.050,而p-ERK1/2表达无明显变化,其灰度值为:0.282±0.005。2.当BV2细胞起保护作用时,梓醇可进一步增强这种保护作用,使PC12细胞存活率进一步增加,而使p-p38MAPK(0.125±0.009,P0.01)与p-JNK(0.077±0.002,P0.01)表达明显减低;当BV2细胞对起损伤作用时,梓醇则明显降低了这种损伤作用,并显著抑制了p-p38MAPK(0.265±0.011,P0.01)与p-JNK、(0.304±0.002,P0.01)的表达。梓醇对p-ERK1/2的表达无论是在损伤还是保护条件下均未见明显变化(0.279±0.020, P0.050 ;0.283±0.023, P0.05)。结论:1.小胶质细胞在不同程度缺氧条件下功能状态的变化与p38MAPK、JNK、ERK1/2磷酸化水平有关,p38MAPK、JNK信号通路参与了损伤作用,而ERK1/2参与了保护作用。 2.缺氧条件下,梓醇可通过抑制小胶质细胞p38MAPK与JNK信号通路发挥神经保护作用。
[Abstract]:Objective: to investigate the changes of MAPK (mitogen-activated protein kinase) signal transduction pathway and the effect of catalpol on it. To explore the mechanism of microglia functional state change and catalpol action mechanism. Methods: BV2 cell line (mouse microglioma) and PC12 cell line (rat pheochromocytoma) were studied. BV2 cells were cultured in different degrees of hypoxia and with or without catalpol, and PC12 cells were cultured in the medium. The survival rate of PC12 cells was measured by CCK-8 method to reflect the function of BV2 cells. Western blot assay was used to detect the MAPK signal transduction pathway JNKP38 MAPK in different functional states of BV2 cells and before and after catalpol addition. Expression and phosphorylation of ERK1/2 protein. Results 1. The expression of protein was expressed by gray value (the gray value was proportional to the expression of protein, when BV2 cells protected neurons. The expression of p-ERK1 / 2 was significantly higher than that of p-p38 MAPK and p-JNK.The gray value of p-ERK1 / 2 was 0.277 卤0.010, respectively. 0.221 卤0.019 and 0.163 卤0.003; The expression of p-p38 MAPK and p-JNK was further enhanced when BV2 cells were injured. The gray values were 0.398 卤0.005 and 0.495 卤0.050, respectively, but the expression of p-ERK1 / 2 did not change significantly. The gray value of catalpol was 0.282 卤0.005.2.When BV2 cells were protected, catalpol could further enhance the protective effect and increase the survival rate of PC12 cells. The expression of p-p38 MAPKN 0.125 卤0.009 P0.01) and p-JNKN 0.077 卤0.002 P0.01) were significantly decreased. Catalpol significantly decreased the damage of BV2 cells and inhibited p-p38 MAPK 0.265 卤0.011 P0.01) and p-JNK. The expression of p-ERK1 / 2 in catalpol was not significantly changed (0.279 卤0.020) under the condition of injury or protection. P0.050; 0.283 卤0.023, P 0.05. Conclusion the changes of functional state of microglia under different degrees of hypoxia are related to the phosphorylation level of p38 MAPK1 / 2 of JNKN ERK1 / 2. JNK signaling pathway is involved in injury and ERK1/2 is involved in protection. 2. Catalpol can play a neuroprotective role by inhibiting p38 MAPK and JNK signaling pathway in microglia.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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