目的:觀察Apelin-13對(duì)脂多糖（Lipopolysaccharides,LPS）誘導(dǎo)N9小膠質(zhì)細(xì)胞鐵沉積的影響及機(jī)制,為神經(jīng)炎癥的防治提供新的思路和線索。方法:1.LPS對(duì)高鐵環(huán)境下N9小膠質(zhì)細(xì)胞鐵沉積的影響。實(shí)驗(yàn)隨機(jī)分為四組:Control組,FAC組,LPS組,FAC+LPS組。FAC組:枸緣酸鐵銨（Ammonium ferric citrate,FAC,100μM）處理N9小膠質(zhì)細(xì)胞24 h。LPS組:脂多糖（2μg/mL）孵育小膠質(zhì)細(xì)胞6 h。FAC+LPS組:先用FAC（100μM）處理細(xì)胞24 h,再用LPS（2μg/mL）孵育細(xì)胞6 h。MTT法檢測(cè)細(xì)胞活力,普魯士藍(lán)染色觀察細(xì)胞內(nèi)鐵沉積,比色法檢測(cè)細(xì)胞內(nèi)總鐵水平;采用流式細(xì)胞術(shù)檢測(cè)小膠質(zhì)細(xì)胞活化標(biāo)志物CD86表達(dá);Western Blot檢測(cè)N9小膠質(zhì)細(xì)胞中iNOS、TNF-α表達(dá)。2.Apelin-13對(duì)高鐵環(huán)境下LPS誘導(dǎo)N9小膠質(zhì)細(xì)胞鐵沉積的影響。實(shí)驗(yàn)隨機(jī)分為四組:FAC組,FAC+Apelin-13組,FAC+LPS組,FAC+Apelin-13+LPS組。Apelin-13處理組先用apelin-13（0... 

【文章來(lái)源】：南華大學(xué)湖南省

【文章頁(yè)數(shù)】：64 頁(yè)

【學(xué)位級(jí)別】：碩士

【部分圖文】：

Apelin-13、FAC和LPS對(duì)N9小膠質(zhì)細(xì)胞活力的影響

南華大學(xué)碩士學(xué)位論文16圖3.3 高鐵環(huán)境下LPS對(duì)N9小膠質(zhì)細(xì)胞鐵沉積的影響Figure. 3.3 Effect of LPS on iron deposition on N9 microglia in high-ironenvironment. N9 microglia were first pretreated with FAC (100 μM) for 24 h and thentreated with LPS (2 μg/mL) for another 6 h. Iron deposition was observed in N9 microgliawith Prussian blue staining (a) and total iron content was quantified using intracellulariron colorimetric assay kit (b). Iron particles were stained blue (200×). Protein levels ofiron-related transporters, DMT1 (c), Fpn1 (d) and FTH1 (e) were determined by Westernblotting. Data were presented as mean ± SD (n=3).*P<0.05,**P<0.01,***P<0.001vs.Control group;##P<0.01,###P<0.001 vs

圖3.4 Apelin-13對(duì)高鐵環(huán)境下LPS誘導(dǎo)的N9小膠質(zhì)細(xì)胞炎癥因子表達(dá)的影響Figure. 3.4. Effects of apelin-13 on proinflammatory cytokines expression inducedby LPS in N9 microglia in high-iron environment. After establishing a high-iron modelin vitro with FAC (100 μM) for 24 h, N9 microglia were pretreated with apelin-13 (0.1μM) for 24 h and then incubated with LPS (2 μg/mL) for another 6 h. The levels ofTNF-α (a) and iNOS (b) were determined by Western blotting. Flow cytometry for thedetermination of the activation marker CD86 of N9 microglia (c). Data were presented asmean ± SD (n=3).*P<0.05,**P<0.01 vs. FAC group;#P<0.05,##P<0.01,###P<0.001 vsFAC+LPS group.3.3.2 Apelin-13改善高鐵環(huán)境下LPS誘導(dǎo)的N9小膠質(zhì)細(xì)胞鐵沉積上述結(jié)果表明LPS加劇了高鐵環(huán)境下N9小膠質(zhì)細(xì)胞內(nèi)鐵沉積。我們接下來(lái)觀察是否apelin-13處理能夠有效改善LPS誘導(dǎo)的胞內(nèi)鐵沉積現(xiàn)象。結(jié)果如圖3.5所示：

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碩士論文
[1]鐵自噬促Sideroflexin1依賴性線粒體鐵超載介導(dǎo)apelin-13誘導(dǎo)心肌細(xì)胞肥大[D]. 吳娣.南華大學(xué) 2016
[2]IL-6對(duì)BV2小膠質(zhì)細(xì)胞系鐵代謝相關(guān)蛋白的影響及其調(diào)控機(jī)制研究[D]. 周仕達(dá).青島大學(xué) 2014



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