Apelin-13抑制IL-6/STAT3途径改善LPS诱导的N9小胶质细胞铁沉积

发布时间：2021-07-14 05:22

　　目的:观察Apelin-13对脂多糖（Lipopolysaccharides,LPS）诱导N9小胶质细胞铁沉积的影响及机制,为神经炎症的防治提供新的思路和线索。方法:1.LPS对高铁环境下N9小胶质细胞铁沉积的影响。实验随机分为四组:Control组,FAC组,LPS组,FAC+LPS组。FAC组:枸缘酸铁铵（Ammonium ferric citrate,FAC,100μM）处理N9小胶质细胞24 h。LPS组:脂多糖（2μg/mL）孵育小胶质细胞6 h。FAC+LPS组:先用FAC（100μM）处理细胞24 h,再用LPS（2μg/mL）孵育细胞6 h。MTT法检测细胞活力,普鲁士蓝染色观察细胞内铁沉积,比色法检测细胞内总铁水平;采用流式细胞术检测小胶质细胞活化标志物CD86表达;Western Blot检测N9小胶质细胞中iNOS、TNF-α表达。2.Apelin-13对高铁环境下LPS诱导N9小胶质细胞铁沉积的影响。实验随机分为四组:FAC组,FAC+Apelin-13组,FAC+LPS组,FAC+Apelin-13+LPS组。Apelin-13处理组先用apelin-13（0... 

【文章来源】：南华大学湖南省

【文章页数】：64 页

【学位级别】：硕士

【部分图文】：

Apelin-13、FAC和LPS对N9小胶质细胞活力的影响

南华大学硕士学位论文16图3.3 高铁环境下LPS对N9小胶质细胞铁沉积的影响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

图3.4 Apelin-13对高铁环境下LPS诱导的N9小胶质细胞炎症因子表达的影响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改善高铁环境下LPS诱导的N9小胶质细胞铁沉积上述结果表明LPS加剧了高铁环境下N9小胶质细胞内铁沉积。我们接下来观察是否apelin-13处理能够有效改善LPS诱导的胞内铁沉积现象。结果如图3.5所示：

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硕士论文
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