本文選題：甲基苯丙胺 + 小膠質(zhì)細(xì)胞�。� 參考：《南京醫(yī)科大學(xué)學(xué)報(自然科學(xué)版)》2017年09期

【摘要】：目的:觀察甲基苯丙胺(methamphetamine,Meth)對原代小膠質(zhì)細(xì)胞的損傷及炎性相關(guān)基因表達(dá)的影響。方法:原代培養(yǎng)SD胎鼠小膠質(zhì)細(xì)胞,利用MTT和原位末端轉(zhuǎn)移酶標(biāo)記技術(shù)(TUNEL)分別檢測Meth引起小膠質(zhì)細(xì)胞活力和凋亡的變化。采用實(shí)時熒光定量聚合酶鏈反應(yīng)法(real-time PCR)觀察Meth對小膠質(zhì)細(xì)胞炎性相關(guān)因子mRNA表達(dá)的影響。ELISA及試劑盒法檢測Meth作用后小膠質(zhì)細(xì)胞白介素(interleukin,IL)-6、腫瘤壞死因子(tumor necrosis factor,TNF)-α、誘導(dǎo)型一氧化氮合酶(inducible nitric oxide synthase,i NOS)的分泌改變。結(jié)果:MTT實(shí)驗(yàn)顯示,Meth降低小膠質(zhì)細(xì)胞活力,呈濃度依賴性,濃度為200μmol/L時與對照組比較,差異有統(tǒng)計學(xué)意義(P0.01)。TUNEL實(shí)驗(yàn)結(jié)果顯示200μmol/L Meth可引起細(xì)胞凋亡,與對照組比較,差異有統(tǒng)計學(xué)意義(P0.05)。q-PCR結(jié)果顯示Meth作用于小膠質(zhì)細(xì)胞24 h可降低IL-24、一氧化氮合酶3(nitric oxide synthase 3,NOS3)表達(dá)水平,上調(diào)Peli3、Sigma受體1(Sigma receptor 1,Sig1-R)、IL-1β、IL-6、Toll樣受體4(Toll like receptor 4,TLR4)的表達(dá)水平,差異有統(tǒng)計學(xué)意義(P0.05)。此外,蛋白水平亦發(fā)現(xiàn),Meth可促進(jìn)IL-6和TNF-α的分泌。結(jié)論:Meth可降低小膠質(zhì)細(xì)胞活力,誘導(dǎo)小膠質(zhì)細(xì)胞凋亡,并引起IL-1β、IL-1R、IL-6、TLR4等炎性因子mRNA表達(dá)變化,促進(jìn)IL-6和TNF-α的分泌,進(jìn)而可能損傷中樞神經(jīng)系統(tǒng),產(chǎn)生神經(jīng)毒性。
[Abstract]:Aim: to investigate the effect of methamphetamine methamphetamine (methamphetamine) on primary microglia injury and inflammatory related gene expression. Methods: primary cultured fetal SD rat microglia cells were cultured. The changes of microglial activity and apoptosis induced by Meth were detected by MTT and in situ terminal transferase labeling technique (Tunel). Real-time PCR was used to observe the effect of Meth on the expression of mRNA of microglial inflammatory related factors. Elisa and kit were used to detect interleukin-6, tumor necrosis factor- TNF- 偽 and tumor necrosis factor (TNF- 偽) in microglial cells after Meth treatment. The secretory changes of inducible nitric oxide synthase I NOS. Results the cell viability was decreased in a concentration-dependent manner, and the difference was significant when the concentration was 200 渭 mol/L. The results of Tunel showed that 200 渭 mol/L Meth could induce cell apoptosis, compared with the control group. The results showed that Meth could decrease the expression of IL-24, nitric oxide synthase (3(nitric oxide synthase 3) and up-regulate the expression of 1(Sigma receptor 1 Sigma receptor 4(Toll like receptor 4 TLR4 in microglia for 24 h, and the difference was statistically significant (P 0 05). In addition, the protein level also found that Meth can promote the secretion of IL-6 and TNF- 偽. ConclusionMeth can decrease the activity of microglia, induce apoptosis of microglia, and induce the expression of mRNA, such as IL-1 尾 -IL-1RL-6IL-6TLR4, and promote the secretion of IL-6 and TNF- 偽, which may damage the central nervous system and produce neurotoxicity.
【作者單位】： 南京醫(yī)科大學(xué)公共衛(wèi)生學(xué)院衛(wèi)生毒理系;南京醫(yī)科大學(xué)第一附屬醫(yī)院急診醫(yī)學(xué)中心;
【基金】：國家自然科學(xué)基金(81673213,81202230) 江蘇省自然科學(xué)基金(BK20151557) 江蘇高校優(yōu)勢學(xué)科建設(shè)工程資助項(xiàng)目
【分類號】：R749.64

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