【摘要】 目的：探究长链非编码RNA（lncRNA） SNHG1调控铁死亡改善HIV-1 gp120 V3环致CHME-5人源小胶质细胞炎症的分子机制。方法：体外培养CHME-5人源小胶质细胞，设立空白组、随机肽段组、HIV-1 gp120V3环组（HIV-1 gp120组）、HIV-1 gp120+shCon组、HIV-1 gp120+SNHG1 sh2组、HIV-1 gp120+SNHG1 sh2+ferrostatin-1（Fer-1；铁死亡抑制剂）组和HIV-1 gp120+SNHG1 sh2+EX527（Sirt1抑制剂）组。随机肽段和gp120 V3环分别处理正常CHME-5细胞24 h；抑制剂预处理SNHG1 sh2细胞2 h后，gp120 V3环处理24 h。ELISA法检测细胞上清中炎症因子水平；Western blot法检测铁死亡相关蛋白[溶质载体家族7成员11（SLC7A11）和谷胱甘肽过氧化物酶4（GPX4）]、Sirt1和p53蛋白表达水平；酶标仪检测细胞内亚铁离子(Fe²⁺)和丙二醛（MDA）含量。结果：（1）ELISA结果显示：较空白组，HIV-1 gp120组炎症因子肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）和IL-1β表达水平显著提高（P<0.05）；较HIV-1 gp120组，HIV-1 gp120+SNHG1 sh2组炎症因子表达水平显著降低（P<0.05）；较HIV-1 gp120+SNHG1 sh2组，HIV-1 gp120+SNHG1 sh2+Fer-1组炎症因子表达水平显著降低（P<0.05）,HIV-1 gp120+SNHG1 sh2+EX527组炎症因子表达水平显著升高（P<0.01）。（2）Western blot结果显示：较空白组，HIV-1 gp120组铁死亡相关蛋白SLC7A11和GPX4表达水平显著下调（P<0.01）；较HIV-1 gp120组，HIV-1 gp120+SNHG1 sh2组SLC7A11和GPX4表达水平显著上调（P<0.01）；较HIV-1 gp120+SNHG1 sh2组，HIV-1 gp120+SNHG1 sh2+Fer-1组SLC7A11和GPX4表达水平明显上调（P<0.05）,HIV-1 gp120+SNHG1 sh2+EX527组SLC7A11和GPX4表达水平显著下调（P<0.01）,p53表达水平显著上调（P<0.05）。（3）较空白组，HIV-1 gp120组Fe²⁺和MDA含量显著提高（P<0.05）；较HIV-1 HIV-1 gp120组，HIV-1 gp120+SNHG1 sh2组Fe²⁺和MDA含量显著降低（P<0.01）；较HIV-1 gp120+SNHG1 sh2组，HIV-1gp120+SNHG1 sh2+Fer-1组Fe²⁺和MDA含量显著降低（P<0.05）,HIV-1 gp120+SNHG1 sh2+EX527组Fe²⁺和MDA含量显著提高（P<0.05）。结论：敲减SNHG1可减轻HIV-1 gp120 V3环介导的小胶质细胞炎症反应，其机制可能是通过Sirt1/p53信号通路调控铁死亡相关信号分子实现的。更多还原

【Abstract】 AIM:To investigate the molecular mechanism of long noncoding RNA （lncRNA） SNHG1 in regulating ferroptosis to alleviate inflammation in CHME-5 human microglia induced by HIV-1 gp120 V3 loop.METHODS:CHME-5 human microglia were cultured in vitro,and were divided into 7 groups:blank group,random peptide group,gp120 V3 loop group （HIV-1 gp120 group）,HIV-1 gp120+shCon group,HIV-1 gp120+SNHG1 sh2 group,HIV-1gp120+SNHG1 sh2+ferrostatin-1 （Fer-1;ferroptosis inhibitor） group,and HIV-1 gp120+SNHG1 sh2+EX527 （Sirt1 inhibitor） group.Normal CHME-5 cells were treated with random peptide or gp120 V3 loop for 24 h.After pretreatment of SNHG1 sh2 cells with inhibitors for 2 h,the cells were then treated with gp120 V3 loop for 24 h.The levels of inflammatory cytokines in the cell supernatants were detected by ELISA.Western blot was used to detect the protein expression levels of solute carrier family 7 member 11 （SLC7A11）,glutathione peroxidase 4 （GPX4）,Sirt1 and p53.Microplate reader was used to detect the levels of intracellular ferrous ion (Fe²⁺) and malondialdehyde （MDA）.RESULTS:（1） The results of ELISA showed that the expression levels of tumor necrosis factor-α （TNF-α）,interleukin-6 （IL-6） and IL-1β in HIV-1gp120 group were significantly higher than those in blank group （P<0.05）.Compared with HIV-1 gp120 group,the expression levels of inflammatory cytokines in HIV-1 gp120+SNHG1 sh2 group were significantly decreased （P<0.05）.Compared with HIV-1 gp120+SNHG1 sh2 group,the expression levels of inflammatory factors in HIV-1 gp120+SNHG1sh2+Fer-1 were significantly decreased （P<0.05）,but those in HIV-1 gp120+SNHG1 sh2+EX527 group were significantly increased （P<0.01）.（2） The results of Western blot showed that compared with blank group,the expression levels of ferroptosis-related proteins SLC7A11 and GPX4 in HIV-1 gp120 group were significantly down-regulated （P<0.01）.Compared with HIV-1 gp120 group,the expression levels of SLC7A11 and GPX4 in HIV-1 gp120+SNHG1 sh2 group were significantly up-regulated （P<0.01）.Compared with HIV-1 gp120+SNHG1 sh2 group,the expression levels of SLC7A11and GPX4 in HIV-1 gp120+SNHG1 sh2+Fer-1 group were significantly up-regulated （P<0.05）,but the expression levels of SLC7A11 and GPX4 in HIV-1 gp120+SNHG1 sh2+EX527 group were significantly down-regulated （P<0.01）,and the expression level of p53 was significantly up-regulated （P<0.05）.（3） Compared with blank group,the levels of Fe²⁺and MDA in HIV-1 gp120 group were significantly increased （P<0.05）.Compared with HIV-1 gp120 group,the levels of Fe²⁺and MDA in HIV-1 gp120+SNHG1 sh2 group were significantly decreased （P<0.01）.Compared with HIV-1 gp120+SNHG1sh2 group,Fe²⁺and MDA in HIV-1 gp120+SNHG1 sh2+Fer-1 group were significantly decreased （P<0.05）,but those in HIV-1 gp120+SNHG1 sh2+EX527 group was significantly increased （P<0.05）.CONCLUSION:Knockdown of SNHG1can attenuate the inflammation in microglia induced by HIV-1 gp120 V3 loop,which may be achieved by regulating ferroptosis-related signaling molecules through the Sirt1/p53 signaling pathway.更多还原