Li Junfen, Zhou Yiqing, He Sicheng, Mao Wenjing, Han Xinbing, Zhang Xinyi, Wang Yan
State Key Laboratory of Ultrasound in Medicine and Engineering, Chongqing Medical University, Chongqing, China; Chongqing Key Laboratory of Biomedical Engineering, Chongqing Medical University, Chongqing, China.
State Key Laboratory of Ultrasound in Medicine and Engineering, Chongqing Medical University, Chongqing, China; Chongqing Key Laboratory of Biomedical Engineering, Chongqing Medical University, Chongqing, China.
Exp Eye Res. 2025 Sep;258:110450. doi: 10.1016/j.exer.2025.110450. Epub 2025 May 29.
To investigate the effect of low-intensity pulsed ultrasound (LIPUS) on epithelial-mesenchymal transition (EMT) in lens epithelial cells. EMT was induced using high glucose (HG) in SRA01/04 cells. Optimal parameters for LIPUS irradiation were determined by cell counting kit-8 assays and flow cytometry. Cell morphology was assessed by light microscopy, while cell migration ability was analyzed by a wound healing assay. Levels of specific proteins and the relationship between autophagy and the cytoskeleton were examined by immunofluorescence (IF) staining and Western blot (WB). Cytoskeletal structures were visualized by phalloidin staining and autophagosomes were quantified by transmission electron microscopy. EMT was successfully induced by HG treatment. Compared to the model group, LIPUS irradiation resulted in a change in cell morphology from spindle to oval, a significant decrease in cell migration area, and an increase in E-cadherin and LC3B/LC3A levels. In contrast, α-SMA and SQSTM1/P62 levels decreased, the number of autophagosomes increased and F-actin levels decreased in the LIPUS group. SRA01/04 cells treated with LIPUS irradiation after autophagy inhibitors 3-MA and CQ showed increased cell migration area compared to the 3-MA/CQ group; LC3B/LC3A levels decreased; SQSTM1/P62 and F-actin levels increased in the LIPUS + 3-MA/CQ group compared to 3-MA/CQ treatment alone. Colocalization of the cytoskeletal marker Arpc2 with the autophagy marker SQSTM1/P62 was also observed. After treatment with the cytoskeletal inhibitor CK666+LIPUS combination therapy, SQSTM1/P62 levels increased while LC3B/LC3A levels decreased. LIPUS inhibited HG-induced EMT by restoring autophagy, which appears to be associated with cytoskeletal remodeling.
研究低强度脉冲超声（LIPUS）对晶状体上皮细胞上皮-间质转化（EMT）的影响。使用高糖（HG）在SRA01/04细胞中诱导EMT。通过细胞计数试剂盒-8检测和流式细胞术确定LIPUS照射的最佳参数。通过光学显微镜评估细胞形态，同时通过伤口愈合试验分析细胞迁移能力。通过免疫荧光（IF）染色和蛋白质免疫印迹（WB）检测特定蛋白水平以及自噬与细胞骨架之间的关系。通过鬼笔环肽染色观察细胞骨架结构，通过透射电子显微镜对自噬体进行定量分析。HG处理成功诱导了EMT。与模型组相比，LIPUS照射导致细胞形态从纺锤形变为椭圆形，细胞迁移面积显著减少，E-钙黏蛋白和LC3B/LC3A水平升高。相反，LIPUS组中α-平滑肌肌动蛋白（α-SMA）和Sequestosome1/P62（SQSTM1/P62）水平降低，自噬体数量增加，F-肌动蛋白水平降低。自噬抑制剂3-甲基腺嘌呤（3-MA）和氯喹（CQ）处理后接受LIPUS照射的SRA01/04细胞与3-MA/CQ组相比，细胞迁移面积增加；LC3B/LC3A水平降低；与单独的3-MA/CQ处理相比，LIPUS + 3-MA/CQ组中SQSTM1/P62和F-肌动蛋白水平升高。还观察到细胞骨架标记物Arpc2与自噬标记物SQSTM1/P62的共定位。在用细胞骨架抑制剂CK666 + LIPUS联合治疗后，SQSTM1/P
