目的 探讨谷氨酸对HT22细胞线粒体自噬和细胞凋亡的影响,并评估虾青素预处理的保护作用及其分子机制。方法 用谷氨酸及虾青素处理HT22细胞,通过蛋白印迹及聚合酶联反应等评估其对线粒体自噬的影响。结果 谷氨酸处理显著抑制线粒体初级自噬（PINK1、Parkin、pULK1^ser555和LC3Ⅱ）和次级自噬（LAMP1和Rab7）,上调cleaved Caspase-3的表达（P<0.05）。虾青素预处理减少细胞凋亡,恢复了线粒体自噬,PINK1、Parkin、pULK1^ser555和LC3Ⅱ的表达水平上升（分别为2.3倍、2.6倍、83.3%及81.1%）（P<0.05）,该作用被自噬抑制剂BafA1阻断。此外,谷氨酸抑制Nrf2核内转移和NLRX1表达,而预处理显著促进Nrf2的核内转移并上调NLRX1,分别上调25.8%、33.2%。生物信息学分析显示NLRX1启动子区域含有3个Nrf2结合位点,提示Nrf2通过调控NLRX1转录活性发挥作用。结论 文章首次揭示虾青素通过Nrf2/NLRX1通路激活线粒体自噬,展现神经保护作用。

Objective To explore the effects of glutamate on mitophagy and apoptosis in HT22 cells and evaluate the protective effects and molecular mechanisms of astaxanthin pretreatment．Methods HT22 cells were treated with glutamate and astaxanthin．The effects on mitophagy were assessed using Western Blot and PCR．Results Glutamate treatment significantly inhibited primary mitophagy（PINK1,Parkin,pULK1^ser555 and LC3II）and secondary mitophagy（LAMP1 and Rab7）while upregulating cleaved Caspase-3 expression．Astaxanthin pretreatment notably reduced apoptosis and restored mitophagy,the expression levels of PINK1,Parkin,pULK1^Ser555 and LC3II were significantly upregulated（by 2.3-fold,2.6-fold,83.3% and 81.1% respectively,P<0.05）,but this effect was blocked by the autophagy inhibitor BafA1．Additionally,glutamate suppressed Nrf2 nuclear translocation and NLRX1 expression,whereas astaxanthin promoted Nrf2 nuclear translocation and increased NLRX1 expression by 25.8% and 33.2%,respectively．Bioinformatics analysis revealed three Nrf2 binding sites in the NLRX1 promoter region,suggesting that Nrf2 may regulate NLRX1 transcriptional activity．Conclusions The study demonstrates that astaxanthin exhibited potential neuroprotective effect by activating mitophagy through the Nrf2/NLRX1 pathway．

目的 评估全身免疫炎症指数（SII）、中性粒细胞/淋巴细胞比值（NLR）和血小板/淋巴细胞比值（PLR）在区分急性缺血性卒中（AIS）伴发卵圆孔未闭（PFO）与非伴发PFO患者的价值。方法 回顾性分析100例AIS患者的血液和血清指标,计算SII、NLR和PLR,使用Logistic回归及受试者操作特征（ROC）曲线分析3项指标在鉴别AIS伴发PFO与非伴发PFO中的价值。结果 伴发PFO的AIS患者SII、NLR、PLR高于非伴发PFO的AIS患者,其中以SII最为明显（P均<0.05）。单因素Logistic回归显示,中性粒细胞计数、淋巴细胞计数、PLR、NLR、SII与AIS伴发PFO有关（P<0.05）。ROC曲线分析结果,SII、NLR、PLR鉴别AIS伴PFO与非伴PFO患者,最佳阈值分别为476.4、1.99、115.3,曲线下面积分别为0.777、0.767、0.708。结论 SII、NLR和PLR可作为鉴别AIS患者是否伴发PFO的生物标志物,具有潜在临床应用价值。

Objective To evaluate the value of systemic immune-inflammatory index（SII）,neutrophil/lymphocyte ratio（NLR）and platelet/lymphocyte ratio（PLR）in distinguishing acute ischemic stroke（AIS）patients with patent foramen ovale（PFO）and without PFO．Methods A retrospective analysis of blood and serum indicators in 100 AIS patients was conducted,and SII,NLR and PLR indices were calculated．Logistic regression and ROC curve analyses were performed．Results SII,NLR and PLR were significantly higher in PFO patients than in non-PFO patients,with SII being the most significant．Univariate logistic regression showed that Neu,Lym,PLR,NLR,and SII variables were significantly associated with AIS combined with PFO（P<0.05）．ROC curve analysis revealed that the optimal cut-off values for SII,NLR and PLR in distinguishing AIS patients with PFO from those without PFO were 476.4,1.99 and 115.3,respectively,with area under the curve of 0.777,0.767 and 0.708．Conclusions SII,NLR and PLR can serve as biomarkers for identifying AIS patients with PFO,offering potential clinical application value．

       目的   探讨谷氨酸对HT22细胞线粒体自噬和细胞凋亡的影响，并评估虾青素预处理的保护作用及其分子机制。方法   用谷氨酸及虾青素处理HT22细胞，通过蛋白印迹及聚合酶联反应等评估其对线粒体自噬的影响。结果   谷氨酸处理显著抑制线粒体初级自噬（PINK1、Parkin、pULK1^ser555和LC3Ⅱ）和次级自噬（LAMP1和Rab7），上调cleaved Caspase-3的表达（P＜0.05）。虾青素预处理减少细胞凋亡，恢复了线粒体自噬，PINK1、Parkin、pULK1^ser555和LC3Ⅱ的表达水平上升（分别为2.3倍、2.6倍、83.3%及81.1%）（P＜0.05），该作用被自噬抑制剂BafA1阻断。此外，谷氨酸抑制Nrf2核内转移和NLRX1表达，而预处理显著促进Nrf2的核内转移并上调NLRX1，分别上调25.8%、33.2%。生物信息学分析显示NLRX1启动子区域含有3个Nrf2结合位点，提示Nrf2通过调控NLRX1转录活性发挥作用。结论   文章揭示虾青素通过Nrf2/NLRX1通路激活线粒体自噬，展现神经保护作用。

       Objective  To explore the effects of glutamate on mitophagy and apoptosis in HT22 cells and evaluate the protective effects and molecular mechanisms of astaxanthin pretreatment．Methods  HT22 cells were treated with glutamate and astaxanthin．The effects on mitophagy were assessed using Western Blot and PCR．Results  Glutamate treatment  significantly inhibited primary mitophagy（PINK1，Parkin，pULK1^ser555 and LC3II）and secondary mitophagy（LAMP1 and Rab7）while upregulating cleaved Caspase-3 expression．Astaxanthin pretreatment notably reduced apoptosis and restored mitophagy，the expression levels of PINK1，Parkin，pULK1^ser555 and LC3II were significantly upregulated（by 2.3-fold，2.6-fold，83.3% and 81.1% respectively，P＜0.05），but this effect was blocked by the autophagy inhibitor BafA1．Additionally，glutamate suppressed Nrf2 nuclear translocation and NLRX1 expression，whereas astaxanthin promoted Nrf2 nuclear translocation and increased NLRX1 expression by 25.8% and 33.2%，respectively．Bioinformatics analysis  revealed three Nrf2 binding sites in the NLRX1 promoter region，suggesting that Nrf2 may regulate NLRX1 transcriptional activity．Conclusions  The study demonstrates that astaxanthin exhibited potential neuroprotective effect by activating mitophagy through the Nrf2/NLRX1 pathway．