激活TREK-TRAAK K2P对氧化损伤人RPE细胞吞噬功能的保护作用

Protective effect of activating TREK-TRAAK K2P on the phagocytic function of oxidative damaged human RPE

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目的:观察弱内向整流相关及花生四烯酸激活的弱内向整流相关双孔钾离子通道(tandem of pore domains in a weak inward rectifying related-tandem of pore domains in a weak inward rectifying related arachidonic acid activated two pore-domain potassium channels,TREK-TRAAK K2P)激活对氧化损伤的人视网膜色素上皮细胞(retinal pigment epithelial, RPE)吞噬功能的影响。方法:免疫荧光法检测TREK-1、TREK-2及TRAAK通道蛋白在人RPE细胞的表达,以不同时间和浓度梯度的叔丁基过氧化氢(tert-butyl hydroperoxide, t-BHP)诱导人RPE细胞氧化损伤。分对照组、t-BHP组、利鲁唑加t-BHP组,利鲁唑组,向各组加入2×107/mL的荧光微球或FITC标记猪感光细胞外节膜盘孵育6 h,固定染色后荧光显微镜拍照,Image-Pro Plus 6.0软件分析计算RPE细胞吞噬率和吞噬指数。结果:TREK-1、TREK-2、TREEK通道蛋白亚型在人RPE细胞质中均高表达。人RPE细胞在t-BHP干预后存活率呈浓度和时间依赖性,200 μmol/L t-BHP干预6 h与对照组存活率比较差异无统计学意义(P>0.05),400μmol/L t-BHP干预6 h致半数死亡。特异性吞噬指数:t-BHP组低于其他各组(P<0.001),而利鲁唑组高于对照组,差异无统计学意义(P>0.05)。特异性吞噬率:t-BHP组低于其他各组,差异无统计学意义(P>0.05)。非特异性吞噬指数:t-BHP组低于其他各组(P<0.001);非特异性吞噬率:t-BHP组低于利鲁唑加t-BHP组,差异无统计学意义(P>0.05),两两比较,仅利鲁唑组高于t-BHP组(P<0.05)。结论:激活TREK-TRAAK K2P通道可保护氧化损伤人RPE细胞的吞噬功能。  
Abstract Objective: To observe the effect of TREK-TRAAK K2P activation on the phagocytic function of oxidative damaged human retinal pigment epithelial cells (RPE). Methods: Immunofluorescence staining was used to detect the expression of TREK-1, TREK-2 and TRAAK channel proteins in human RPE cells, and the RPE cell oxidative damage model was induced by tert-butyl hydroperoxide (t-BHP) at different time and concentration gradients. The study was divided into control group, t-BHP model group, riluzole plus t-BHP group, and riluzole group. 2×107/mL fluorescent microspheres or FITC-labeled porcine photoreceptor outer segment membrane discs were added to each group for 6h incubation, fluorescence photos were obtained after fixation and staining, and the phagocytosis rate and phagocytosis index of RPE cells was analyzed and calculated by Image-Pro Plus 6.0 software. Results: TREK-1, TREK-2, and TREEK channel subtype proteins were all highly expressed in the human RPE cytoplasm. The survival rate of human RPE cells after t-BHP intervention was concentration- and time-dependent. There was no statistical difference in the survival rate between 200 μmol/L t-BHP intervention for 6 hours and the control group (P>0.05), and 400 μmol/L t-BHP intervention for 6 hours induced half death. Specific phagocytic index: The t-BHP group was lower than other groups (P<0.001). Specific phagocytosis rate: the t-BHP group was lower than other groups, with no statistical difference (P>0.05). Nonspecific phagocytic index: the t-BHP group was lower than other groups (P<0.001); nonspecific phagocytic rate: the t-BHP model group was lower than the riluzole plus t-BHP group, with no statistical difference (P>0.05), and only the riluzole group was higher than the t-BHP model group (P<0.05) in pairwise comparison. Conclusions: Activation of TREK-TRAAK K2P can protect the phagocytic function of human RPE cells damaged by oxidative stress.
论著

内界膜翻瓣术联合空气填充治疗黄斑裂孔的效果评价

Efficacy of macular hole surgery with inverted internal limiting membrane flap technique and air tamponade

:323-330
 
目的:评估内界膜翻瓣术联合空气填充在黄斑裂孔手术应用中的有效性。方法:回顾性分析中山眼科中心2016—2019年入院,因黄斑裂孔行内界膜翻瓣术联合空气填充术治疗的患者71例71眼,术后俯卧位1~3 d且随访达6个月以上。通过OCT判断患者黄斑裂孔闭合率,比较患者术前及术后最佳矫正视力(best corrected visual acuity, BCVA),通过MAIA微视野计检查评估术前与术后黄斑平均光敏度、黄斑完整指数、固视稳定性P1及P2等指标。结果:在总的71只眼中,有91.5%达到了黄斑裂孔的完全闭合。患者术前及术后BCVA提升(P<0.001),黄斑平均光敏度增加(P<0.000 1)、黄斑完整指数降低(P<0.000 1)、固视稳定性P1(P<0.000 1)及P2(P<0.001)均提升。结论:内界膜翻瓣术联合空气填充是治疗黄斑裂孔有效的手术方式,空气填充可有效地辅助黄斑裂孔愈合,可作为优先考虑的术中填充物,减轻患者因长时间保持面向下体位带来的痛苦。微视野计参数可用作评估黄斑结构及功能恢复的良好指标。
Objective: To assess the efficacy of the inverted internal limiting membrane (ILM) flap technique with air tamponade for macular holes (MHs). Methods: A retrospective analysis was conducted on 71 patients (71 eyes) who underwent inverted ILM flap surgery combined with air tamponade for macular holes at Zhongshan Ophthalmic Center from 2016 to 2019. Patients were positioned face down for 1-3 days postoperatively and followed up for more than 6 months. OCT was used to determine the closure rate of macular holes, and the best corrected visual acuity (BCVA) before and after surgery was compared. The MAIA microperimeter was used to assess the average macular sensitivity, macular integrity index, and fixation stability P1 and P2 before and after surgery. Results: Of the total 71 eyes, 91.5% achieved complete MH closure. The best corrected visual acuity (BCVA) significantly improved before and after surgery (< 0.001), average macular sensitivity significantly increased (P < 0.000 1), macular integrity index significantly decreased (P < 0.000 1), and fixation stability P1 (P < 0.000 1) and P2 (P < 0.001) both significantly improved. Conclusions: Inverted ILM flap technique combined with air tamponade provides an effective approach for the management of MHs. The air tamponade should be considered as the preferred in MH surgeries, reducing the discomfort caused by prolonged face-down positioning. Parameters in microperimetry can serve as good indicators for assessing the recovery of macular structure and function.
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  • 眼科学报

    主管:中华人民共和国教育部
    主办:中山大学
    承办:中山大学中山眼科中心
    主编:林浩添
    主管:中华人民共和国教育部
    主办:中山大学
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  • Eye Science

    主管:中华人民共和国教育部
    主办:中山大学
    承办:中山大学中山眼科中心
    主编:林浩添
    主管:中华人民共和国教育部
    主办:中山大学
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