【摘要】：目的：觀察p38MAPK信號(hào)通路變化對小膠質(zhì)細(xì)胞功能及細(xì)胞因子分泌的影響，探討p38MAPK信號(hào)通路在小膠質(zhì)細(xì)胞功能變化中的作用機(jī)制。 方法：以BV2細(xì)胞株（小鼠小膠質(zhì)細(xì)胞瘤細(xì)胞）和PC12細(xì)胞株（大鼠嗜鉻細(xì)胞瘤細(xì)胞）為研究對象，，分別用其來代表小膠質(zhì)細(xì)胞和神經(jīng)元。在不同程度缺氧條件下培養(yǎng)BV2細(xì)胞，再用其培養(yǎng)液培養(yǎng)PC12細(xì)胞。通過CCK-8法測定PC12細(xì)胞存活率以此來評價(jià)BV2細(xì)胞功能；應(yīng)用ELISA法檢測不同狀態(tài)下BV2細(xì)胞分泌TNFα、NGF和IL-1β的情況。在低氧培養(yǎng)BV2細(xì)胞的條件下將p38MAPK信號(hào)通路阻斷劑（SB203580）加入到BV2細(xì)胞培養(yǎng)液中，測定阻斷p38MAPK信號(hào)通路后BV2細(xì)胞的功能變化及TNFα、IL-1β、NGF的分泌情況。 結(jié)果：1、BV2細(xì)胞低氧培養(yǎng)1h分泌NGF、TNF-α和IL-1β的量分別為；215.81±13.11pg/ml、808.91±7.41pg/ml和7.89±0.47pg/ml，阻斷p38MAPK信號(hào)通路后NGF分泌增多為376.25±11.67pg/ml，TNF-α和IL-1β分泌降低分別是544.85±8.13pg/ml和4.75±0.23pg/ml(p0.01)。低氧培養(yǎng)12h分泌NGF、TNF-α和IL-1β的量分別為：142.35±1.60pg/ml、1569.27±19.07pg/ml和28.53±6.2pg/ml，阻斷p38MAPK信號(hào)通路后TNF-α分泌量下降為1127.88±14.03pg/ml，NGF分泌量升高為166.40±3.74pg/ml(p0.01)，而IL-1β的分泌量無明顯變化(p0.05)。 2、正常對照組PC12細(xì)胞存活率若按100%計(jì)算，損傷對照組細(xì)胞存活率為48.2%，低氧1h組細(xì)胞存活率為71.4%。與低氧1h組相比阻斷p38MAPK信號(hào)通路后PC12細(xì)胞存活率上升，存活率為86.9%（p0.01）。低氧12h組細(xì)胞存活率為29.6%。與低氧12h組相比阻斷p38MAPK信號(hào)通路后PC12細(xì)胞存活率上升，為51.4%（p0.01）。 結(jié)論：1、p38MAPK信號(hào)通路參與了小膠質(zhì)細(xì)胞的損傷性作用，阻斷p38MAPK信號(hào)通路小膠質(zhì)細(xì)胞的保護(hù)性作用增強(qiáng)。 2、p38MAPK信號(hào)通路參與調(diào)控小膠質(zhì)細(xì)胞的NGF、TNF-α的分泌表達(dá)，阻斷p38MAPK信號(hào)通路NGF分泌增多、TNF-α分泌減少。
[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 偽, 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 偽, IL-1 尾 and NGF were measured after blocking p38MAPK signaling pathway. Results: 1 the levels of NGF,TNF- 偽 and IL-1 尾 secreted by BV2 cells were 215.81 鹵13.11pg / ml 808.91 鹵7.41pg/ml and 7.89 鹵0.47pg / ml, respectively. After blocking the p38MAPK signaling pathway, the secretion of NGF increased to 376.25 鹵11.67pgml / ml and the secretion of IL-1 尾 decreased by 544.85 鹵8.13pg/ml and 4.75 鹵0.23pg/ml, respectively. The secretion of NGF,TNF- 偽 and IL-1 尾 were 142.35 鹵1.60pg / ml 1569.27 鹵19.07pg/ml and 28.53 鹵6.2pg / ml, respectively. After blocking the p38MAPK signaling pathway, the secretion of TNF- 偽 decreased to 1127.88 鹵14.03pgml / ml, and the secretion of IL-1 尾 increased to 166.40 鹵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- 偽 in microglia. The secretion of NGF increased and the secretion of TNF- 偽 decreased after blocking the p38MAPK signaling pathway.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R329

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