本文選題：甲基苯丙胺　切入點(diǎn)：小膠質(zhì)細(xì)胞　出處：《南京醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的觀察甲基苯丙胺（Methamphetamine，Meth）對(duì)大鼠胎鼠小膠質(zhì)細(xì)胞的損傷作用。方法原代培養(yǎng)SD胎鼠小膠質(zhì)細(xì)胞，細(xì)胞計(jì)數(shù)試劑盒（CCK-8及MTT）和原位末端轉(zhuǎn)移酶標(biāo)記技術(shù)（TUNEL）分別檢測(cè)Meth引起小膠質(zhì)細(xì)胞活力和凋亡的變化。結(jié)果MTT實(shí)驗(yàn)顯示，甲基苯丙胺呈濃度依賴性（10、30、100、300、1000μM）降低小膠質(zhì)細(xì)胞活力。在Meth濃度為10、30、100μM時(shí)，細(xì)胞活力有所下降，當(dāng)濃度為300、1000μM時(shí)，Meth會(huì)造成細(xì)胞活力下降，差別有統(tǒng)計(jì)學(xué)差異（p0.05）。CCK-8實(shí)驗(yàn)，鉀離子通道非特異性抑制劑Tetraethylamine（TEA）、4-Aminopyridine（4-AP）及鉀離子通道亞型Kv1.3特異性抑制劑Margatoxin（MgTx）預(yù)孵后再加入Meth，Meth所引起的細(xì)胞損傷可被部分逆轉(zhuǎn)。TUNEL實(shí)驗(yàn)可知，，100、300μM Meth可引起細(xì)胞凋亡，300μM時(shí)差異有統(tǒng)計(jì)學(xué)意義。結(jié)論Meth明顯引起小膠質(zhì)細(xì)胞損傷，該損傷過(guò)程可以部分被TEA、4-AP及MgTx等鉀離子通道抑制劑逆轉(zhuǎn)。 目的觀察甲基苯丙胺（Meth）引起小膠質(zhì)細(xì)胞損傷中電壓門控鉀離子通道亞型Kv1.3、Kv1.5的作用。方法原代培養(yǎng)SD胎鼠小膠質(zhì)細(xì)胞，采用蛋白質(zhì)印跡法（western blot）和實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)法（RT-PCR）觀察Kv1.3、Kv1.5表達(dá)變化情況。采用RT-PCR法觀察Meth作用后小膠質(zhì)細(xì)胞內(nèi)炎性因子分泌情況。同時(shí)探討Meth引起小膠質(zhì)細(xì)胞內(nèi)Kv1.3變化及Meth與p38-絲裂原活化蛋白激酶（MAPK）信號(hào)通路之間的關(guān)系。結(jié)果Meth引起Kv1.3mRNA和蛋白水平表達(dá)增高，Kv1.3特異性抑制劑MgTx可減少上述表達(dá)。Meth對(duì)Kv1.5mRNA和蛋白水平表達(dá)均無(wú)明顯影響。Meth所致小膠質(zhì)細(xì)胞內(nèi)炎性因子表達(dá)量的增高亦可被MgTx所阻滯。Meth引起的小膠質(zhì)細(xì)胞損傷可通過(guò)預(yù)孵MgTx降低，其損傷可以激活p38MAPK通路。結(jié)論Kv1.3可能參與甲基苯丙胺引起小膠質(zhì)細(xì)胞損傷機(jī)，該通道抑制后，Meth引起的炎性因子表達(dá)顯著降低。此外，Meth明顯激活p38MAPK通路。因而，Kv1.3可能成為小膠質(zhì)細(xì)胞損傷治療的一個(gè)新靶點(diǎn)。
[Abstract]:Objective to observe the damage effect of methamphetamine on rat fetal microglia. Methods Primary culture of SD fetal rat microglia cells was performed. Cell count kit CCK-8 and MTT) and in situ terminal transferase labeling (Tunel) were used to detect the changes of microglia viability and apoptosis induced by Meth. Methamphetamine decreased the viability of microglia cells in a dose-dependent manner. When the concentration of Meth was 10 ~ 30100 渭 M, the cell viability was decreased, and when the concentration was 300 渭 M, the cell viability was decreased, and the difference was statistically significant (p 0.05N. CCK-8). Tetraethylamineine (Tetraethylamine), a nonspecific inhibitor of potassium channel, Tetraethylamine, 4-Aminopyridine 4-AP), and the potassium channel subtype Kv1.3 specific inhibitor, Kv1.3, were preincubated and then added with Methanemeth, which could be partially reversed. Tunel showed that 100300 渭 M Meth could induce apoptosis at 300 渭 M. Conclusion Meth can obviously induce microglia injury. This process can be partially reversed by potassium channel inhibitors such as tea 4-AP and MgTx. Objective to observe the effect of voltage-gated potassium channel subtype Kv1.3 Kv1.5 on microglia injury induced by methamphetamine method primary cultured SD fetal rat microglia cells. Western blot and RT PCR were used to observe the expression of Kv1.3 and Kv1.5. RT-PCR was used to observe the secretion of inflammatory factors in microglia after Meth treatment. Meanwhile, microglia were induced by Meth. The changes of Kv1.3 in cells and the relationship between Meth and p38 mitogen-activated protein kinase (p38 mitogen activated protein kinase) signal pathway. Results Meth induced the increase of Kv1.3mRNA and protein expression. MgTx, a specific inhibitor of Kv1.3, could reduce the expression of Kv1.5mRNA and protein. No significant effect was found on the expression of inflammatory cytokines in microglial cells induced by. Meth. Meth could also be blocked by MgTx. Meth-induced microglial injury could be reduced by preincubation of MgTx. Conclusion Kv1.3 may be involved in the mechanism of microglia injury induced by methamphetamine. The expression of inflammatory cytokines induced by Meth was significantly decreased after this channel was inhibited. In addition, Meth significantly activated the p38MAPK pathway. Therefore, Kv1.3 may be a new target for the treatment of microglia injury.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 徐春華;戴閩;劉遠(yuǎn)厚;;動(dòng)脈粥樣硬化兔血管平滑肌鈣激活鉀通道活性和α亞單位表達(dá)的變化[J];四川醫(yī)學(xué);2008年07期

2 劉云;馬英;;苯丙胺類物質(zhì)濫用對(duì)中樞神經(jīng)系統(tǒng)影響的研究進(jìn)展[J];中國(guó)藥物依賴性雜志;2008年01期

3 曹陽(yáng);張世忠;趙淑琴;;心肌保護(hù)新靶點(diǎn)——線粒體鈣激活鉀通道研究進(jìn)展[J];醫(yī)學(xué)與哲學(xué)(臨床決策論壇版);2009年12期

本文編號(hào)：1646094