p38MAPK信号通路对小胶质细胞功能及NGF、TNFα分泌的影响
发布时间：2018-10-10 20:50
【摘要】：目的：观察p38MAPK信号通路变化对小胶质细胞功能及细胞因子分泌的影响，探讨p38MAPK信号通路在小胶质细胞功能变化中的作用机制。 方法：以BV2细胞株（小鼠小胶质细胞瘤细胞）和PC12细胞株（大鼠嗜铬细胞瘤细胞）为研究对象，，分别用其来代表小胶质细胞和神经元。在不同程度缺氧条件下培养BV2细胞，再用其培养液培养PC12细胞。通过CCK-8法测定PC12细胞存活率以此来评价BV2细胞功能；应用ELISA法检测不同状态下BV2细胞分泌TNFα、NGF和IL-1β的情况。在低氧培养BV2细胞的条件下将p38MAPK信号通路阻断剂（SB203580）加入到BV2细胞培养液中，测定阻断p38MAPK信号通路后BV2细胞的功能变化及TNFα、IL-1β、NGF的分泌情况。 结果：1、BV2细胞低氧培养1h分泌NGF、TNF-α和IL-1β的量分别为；215.81±13.11pg/ml、808.91±7.41pg/ml和7.89±0.47pg/ml，阻断p38MAPK信号通路后NGF分泌增多为376.25±11.67pg/ml，TNF-α和IL-1β分泌降低分别是544.85±8.13pg/ml和4.75±0.23pg/ml(p0.01)。低氧培养12h分泌NGF、TNF-α和IL-1β的量分别为：142.35±1.60pg/ml、1569.27±19.07pg/ml和28.53±6.2pg/ml，阻断p38MAPK信号通路后TNF-α分泌量下降为1127.88±14.03pg/ml，NGF分泌量升高为166.40±3.74pg/ml(p0.01)，而IL-1β的分泌量无明显变化(p0.05)。 2、正常对照组PC12细胞存活率若按100%计算，损伤对照组细胞存活率为48.2%，低氧1h组细胞存活率为71.4%。与低氧1h组相比阻断p38MAPK信号通路后PC12细胞存活率上升，存活率为86.9%（p0.01）。低氧12h组细胞存活率为29.6%。与低氧12h组相比阻断p38MAPK信号通路后PC12细胞存活率上升，为51.4%（p0.01）。 结论：1、p38MAPK信号通路参与了小胶质细胞的损伤性作用，阻断p38MAPK信号通路小胶质细胞的保护性作用增强。 2、p38MAPK信号通路参与调控小胶质细胞的NGF、TNF-α的分泌表达，阻断p38MAPK信号通路NGF分泌增多、TNF-α分泌减少。
[Abstract]:Aim: to investigate the effects of changes of p38MAPK signaling pathway on microglial function and cytokine secretion, and to explore the mechanism of p38MAPK signaling pathway in microglial function change. Methods: BV2 cell lines (mouse microglioma cells) and PC12 cell lines (rat pheochromocytoma cells) were used to represent microglia and neurons respectively. BV2 cells were cultured in different degrees of hypoxia and PC12 cells were cultured in the medium. The function of BV2 cells was evaluated by CCK-8 assay, and the secretion of TNF 伪, NGF and IL-1 尾 by BV2 cells in different states was detected by ELISA assay. P38MAPK signal pathway blocker (SB203580) was added to the culture medium of BV2 cells cultured in hypoxia. The function of BV2 cells and the secretion of TNF 伪, IL-1 尾 and NGF were measured after blocking p38MAPK signaling pathway. Results: 1 the levels of NGF,TNF- 伪 and IL-1 尾 secreted by BV2 cells were 215.81 卤13.11pg / ml 808.91 卤7.41pg/ml and 7.89 卤0.47pg / ml, respectively. After blocking the p38MAPK signaling pathway, the secretion of NGF increased to 376.25 卤11.67pgml / ml and the secretion of IL-1 尾 decreased by 544.85 卤8.13pg/ml and 4.75 卤0.23pg/ml, respectively. The secretion of NGF,TNF- 伪 and IL-1 尾 were 142.35 卤1.60pg / ml 1569.27 卤19.07pg/ml and 28.53 卤6.2pg / ml, respectively. After blocking the p38MAPK signaling pathway, the secretion of TNF- 伪 decreased to 1127.88 卤14.03pgml / ml, and the secretion of IL-1 尾 increased to 166.40 卤3.74pg/ml (p0.01), while the secretion of IL-1 尾 did not change (p0.05). 2. The survival rate of PC12 cells in the control group was 100%. The cell survival rate was 48.2 in the injured control group and 71.4 in the hypoxia 1 hour group. Compared with hypoxia for 1 h, the survival rate of PC12 cells was 86.9% (p0.01) after blocking the p38MAPK signaling pathway. The cell survival rate of 12 h hypoxia group was 29.6%. Compared with 12 h hypoxia group, the survival rate of PC12 cells increased to 51.4% (p0.01) after blocking the p38MAPK signaling pathway. Conclusion: 1 the p38 MAPK signaling pathway is involved in the injury of microglia, and the protective effect of blocking the p38MAPK signaling pathway is enhanced. 2 the p38 MAPK signaling pathway is involved in regulating the secretion of NGF,TNF- 伪 in microglia. The secretion of NGF increased and the secretion of TNF- 伪 decreased after blocking the p38MAPK signaling pathway.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
本文编号：2263199
[Abstract]:Aim: to investigate the effects of changes of p38MAPK signaling pathway on microglial function and cytokine secretion, and to explore the mechanism of p38MAPK signaling pathway in microglial function change. Methods: BV2 cell lines (mouse microglioma cells) and PC12 cell lines (rat pheochromocytoma cells) were used to represent microglia and neurons respectively. BV2 cells were cultured in different degrees of hypoxia and PC12 cells were cultured in the medium. The function of BV2 cells was evaluated by CCK-8 assay, and the secretion of TNF 伪, NGF and IL-1 尾 by BV2 cells in different states was detected by ELISA assay. P38MAPK signal pathway blocker (SB203580) was added to the culture medium of BV2 cells cultured in hypoxia. The function of BV2 cells and the secretion of TNF 伪, IL-1 尾 and NGF were measured after blocking p38MAPK signaling pathway. Results: 1 the levels of NGF,TNF- 伪 and IL-1 尾 secreted by BV2 cells were 215.81 卤13.11pg / ml 808.91 卤7.41pg/ml and 7.89 卤0.47pg / ml, respectively. After blocking the p38MAPK signaling pathway, the secretion of NGF increased to 376.25 卤11.67pgml / ml and the secretion of IL-1 尾 decreased by 544.85 卤8.13pg/ml and 4.75 卤0.23pg/ml, respectively. The secretion of NGF,TNF- 伪 and IL-1 尾 were 142.35 卤1.60pg / ml 1569.27 卤19.07pg/ml and 28.53 卤6.2pg / ml, respectively. After blocking the p38MAPK signaling pathway, the secretion of TNF- 伪 decreased to 1127.88 卤14.03pgml / ml, and the secretion of IL-1 尾 increased to 166.40 卤3.74pg/ml (p0.01), while the secretion of IL-1 尾 did not change (p0.05). 2. The survival rate of PC12 cells in the control group was 100%. The cell survival rate was 48.2 in the injured control group and 71.4 in the hypoxia 1 hour group. Compared with hypoxia for 1 h, the survival rate of PC12 cells was 86.9% (p0.01) after blocking the p38MAPK signaling pathway. The cell survival rate of 12 h hypoxia group was 29.6%. Compared with 12 h hypoxia group, the survival rate of PC12 cells increased to 51.4% (p0.01) after blocking the p38MAPK signaling pathway. Conclusion: 1 the p38 MAPK signaling pathway is involved in the injury of microglia, and the protective effect of blocking the p38MAPK signaling pathway is enhanced. 2 the p38 MAPK signaling pathway is involved in regulating the secretion of NGF,TNF- 伪 in microglia. The secretion of NGF increased and the secretion of TNF- 伪 decreased after blocking the p38MAPK signaling pathway.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
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