纳米凝胶搭载的siRNA通过靶向抑制施万细胞铁死亡促进周围神经损伤修复-《中国医学物理学杂志》

文章信息/Info

Title:: siRNA delivered by nanogels facilitates peripheral nerve repair through inhibiting ferroptosis in Schwann cells

作者:: 杨俊1; 2; 黄欣林3; 覃汉俊4; 谢思远5; 朱煜华6; 吴珺7; 1.中国人民解放军陆军第七十四集团军医院（原解放军第四二一医院）骨科，广东广州 510318； 2.南方医科大学南方医院创伤骨科，广东广州 510515； 3.南方医科大学南方医院关节与骨病外科，广东广州 510515； 4.深圳市人民医院（南方科技大学第一附属医院）创伤骨科，广东深圳 518020； 5.南方医科大学中医药学院，广东广州 510515； 6.南方医科大学南方医院脊柱骨科，广东广州 510515； 7.中国人民解放军陆军第七十四集团军医院（原解放军第四二一医院）门诊部体检中心，广东广州 510318

Author(s):: YANG Jun1; 2; HUANG Xinlin3; QIN Hanjun4; XIE Siyuan5; ZHU Yuhua6; WU Jun7; 1. Department of Orthopedics, the 74th Group Army Hospital of PLA (formerly, the 421st Hospital of PLA), Guangzhou 510318, China 2. Department of Orthopedic Trauma, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China 3. Department of Joint and Bone Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China 4. Department of Orthopedic Trauma, Shenzhen Peoples Hospital (the First Affiliated Hospital of Southern University of Science and Technology), Shenzhen 518020, China 5. School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China 6. Department of Spinal Orthopedics, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China 7. Physical Examination Center of the Outpatient Department, the 74th Group Army Hospital of PLA (formerly, the 421st Hospital of PLA), Guangzhou 510318, China

Keywords:: Keywords: peripheral nerve injury Schwann cell ferroptosis nanogel siRNA targeted therapy

摘要:: 目的：探究周围神经损伤（PNI）后施万细胞的死亡机制，使用纳米凝胶搭载的siRNA靶向抑制施万细胞死亡。方法：下载GEO数据库中PNI的转录组数据，依次进行数据处理、差异分析、GO功能富集分析、铁死亡通路鉴定以及靶基因的筛选。通过蛋白印记和qPCR实验验证靶基因的差异性。自组装单宁酸（TA）-siRNA纳米凝胶，通过丁达尔效应实验、粒径和电位测量、细胞吞噬实验、细胞骨架染色和CCK-8细胞活力实验鉴定纳米凝胶的物理学和生物学特性。通过划痕实验、免疫荧光染色实验、蛋白印记和qPCR实验验证TA-siRNA纳米凝胶抑制施万细胞铁死亡的有效性。结果：PNI后施万细胞出现明显的铁死亡现象，Bex1是调控施万细胞铁死亡的关键基因。TA-siRNA纳米凝胶具有优良的物理学和生物学特性，能够将siRNA成功地携带到损伤的施万细胞中并沉默靶基因，从而有效地抑制施万细胞损伤后的铁死亡。结论：纳米凝胶搭载的siRNA可以靶向抑制施万细胞铁死亡，为临床治疗PNI提供新的方向。

Abstract:: Abstract: Objective To investigate the mechanism of Schwann cell death after peripheral nerve injury (PNI), and to inhibit Schwann cell death by nanogel-delivered siRNA. Methods The transcriptome data of PNI in GEO database were downloaded for data processing, followed by difference analysis, GO functional enrichment analysis, ferroptosis pathway identification, and target gene screening. The differences of target genes were verified through protein imprinting and qPCR experiment. The physical and biological properties of self-assembled tannic acid (TA)-siRNA nanogels were identified by Tyndall effect, particle size and potential measurements, phagocytosis assay, cytoskeleton staining, and CCK-8 cell viability assay. The effectiveness of TA-siRNA nanogel in inhibiting ferroptosis in Schwann cells was verified by scratch experiment, immunofluorescence staining, protein imprinting, and qPCR experiment. Results After PNI, obvious ferroptosis was observed in Schwann cells, and Bex1 was identified as the key gene regulating ferroptosis in Schwann cells. With excellent physical and biological properties, TA-siRNA nanogel could successfully carry siRNA into damaged Schwann cells, and silence their ferroptosis genes, thus effectively inhibiting ferroptosis after Schwann cell damage. Conclusion Nanogel-delivered siRNA can inhibit ferroptosis in Schwann cells, providing a new direction for PNI treatment.