本文關(guān)鍵詞：小膠質(zhì)細(xì)胞不同功能狀態(tài)下MAPK信號(hào)通路變化及梓醇對(duì)其影響　出處：《大連醫(yī)科大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 小膠質(zhì)細(xì)胞 功能狀態(tài) MAPK 梓醇

【摘要】：目的:在前期研究工作基礎(chǔ)上進(jìn)一步觀察小膠質(zhì)細(xì)胞不同功能狀態(tài)下MAPK(絲裂原活化蛋白激酶)信號(hào)轉(zhuǎn)導(dǎo)通路變化及梓醇對(duì)其影響,探討小膠質(zhì)細(xì)胞功能狀態(tài)變化機(jī)制及梓醇作用機(jī)制。 方法:以BV2細(xì)胞株(小鼠小膠質(zhì)細(xì)胞瘤)和PC12細(xì)胞株(大鼠嗜鉻細(xì)胞瘤)為研究對(duì)象,分別用其來代表小膠質(zhì)細(xì)胞和神經(jīng)元。在不同程度缺氧及有無梓醇存在條件下培養(yǎng)BV2細(xì)胞,用其培養(yǎng)液培養(yǎng)PC12細(xì)胞,通過CCK-8法測(cè)定PC12細(xì)胞存活率并以此來反映BV2細(xì)胞功能,同步應(yīng)用Western blot法檢測(cè)BV2細(xì)胞不同功能狀態(tài)下及加入梓醇前后MAPK信號(hào)轉(zhuǎn)導(dǎo)通路JNK、p38MAPK、ERK1/2蛋白表達(dá)及磷酸化水平。 結(jié)果:1.蛋白表達(dá)量應(yīng)用灰度值來表示(灰度值與蛋白表達(dá)量成正比),當(dāng)BV2細(xì)胞對(duì)神經(jīng)元起保護(hù)作用時(shí),p-ERK1/2表達(dá)明顯高于p-p38MAPK與p-JNK,其灰度值分別為:0.277±0.010、0.221±0.019和0.163±0.003;當(dāng)BV2細(xì)胞對(duì)神經(jīng)元起損傷作用時(shí),p-p38MAPK與p-JNK表達(dá)較保護(hù)作用時(shí)進(jìn)一步增強(qiáng),灰度值分別為:0.398±0.005和0.495±0.050,而p-ERK1/2表達(dá)無明顯變化,其灰度值為:0.282±0.005。2.當(dāng)BV2細(xì)胞起保護(hù)作用時(shí),梓醇可進(jìn)一步增強(qiáng)這種保護(hù)作用,使PC12細(xì)胞存活率進(jìn)一步增加,而使p-p38MAPK(0.125±0.009,P0.01)與p-JNK(0.077±0.002,P0.01)表達(dá)明顯減低;當(dāng)BV2細(xì)胞對(duì)起損傷作用時(shí),梓醇則明顯降低了這種損傷作用,并顯著抑制了p-p38MAPK(0.265±0.011,P0.01)與p-JNK、(0.304±0.002,P0.01)的表達(dá)。梓醇對(duì)p-ERK1/2的表達(dá)無論是在損傷還是保護(hù)條件下均未見明顯變化(0.279±0.020, P0.050 ;0.283±0.023, P0.05)。 結(jié)論:1.小膠質(zhì)細(xì)胞在不同程度缺氧條件下功能狀態(tài)的變化與p38MAPK、JNK、ERK1/2磷酸化水平有關(guān),p38MAPK、JNK信號(hào)通路參與了損傷作用,而ERK1/2參與了保護(hù)作用。 2.缺氧條件下,梓醇可通過抑制小膠質(zhì)細(xì)胞p38MAPK與JNK信號(hào)通路發(fā)揮神經(jīng)保護(hù)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 鹵0.010, respectively. 0.221 鹵0.019 and 0.163 鹵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 鹵0.005 and 0.495 鹵0.050, respectively, but the expression of p-ERK1 / 2 did not change significantly. The gray value of catalpol was 0.282 鹵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 鹵0.009 P0.01) and p-JNKN 0.077 鹵0.002 P0.01) were significantly decreased. Catalpol significantly decreased the damage of BV2 cells and inhibited p-p38 MAPK 0.265 鹵0.011 P0.01) and p-JNK. The expression of p-ERK1 / 2 in catalpol was not significantly changed (0.279 鹵0.020) under the condition of injury or protection. P0.050; 0.283 鹵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.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 郝玉通;陳純海;楊學(xué)森;余爭(zhēng)平;;JAK2/STAT3信號(hào)通路參與脂多糖誘導(dǎo)小膠質(zhì)細(xì)胞活化的研究[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2009年12期

2 張桂蓮;姚麗;杜峗;張茹;卜寧;劉t熃，

本文編號(hào)：1373519