小鼠骨髓间质干细胞来源的胞外囊泡对尼古丁诱导的小鼠生精细胞损伤的修复作用

目的: 构建尼古丁诱导的小鼠生精细胞体外损伤模型,探讨小鼠骨髓间质干细胞来源的胞外囊泡(mouse bone marrow mesenchymal stem cell-derived extracellular vesicles, mBMMSCs-EVs)对于小鼠GC-1生精细胞损伤的修复作用。方法:采用不同浓度尼古丁(0、8、16、32 μg/mL)处理GC-1生精细胞24、48、72 h,通过MTT实验筛选尼古丁诱导GC-1生精细胞损伤的最适浓度和时间；将GC-1生精细胞分为对照组、尼古丁组、尼古丁+胞外囊泡组；通过MTT实验、Transwell实验和流式细胞术凋亡实验观察mBMMSCs-EVs干预前后受损生精细胞GC-1生物学特性的变化。结果:MTT实验结果表明,32 μg/mL尼古丁作用GC-1生精细胞24 h时,细胞增殖能力明显降低(P<0.05),mBMMSCs-EVs作用于尼古丁预处理后,细胞增殖能力明显增高(P<0.05）；与对照组相比,尼古丁组GC-1生精细胞迁移能力明显降低,细胞凋亡能力明显升高(P均<0.01),与尼古丁组相比,尼古丁+胞外囊泡组生精细胞增殖和迁移能力明显升高,细胞凋亡能力明显下降(P<0.01或<0.05)。结论:mBMMSCs-EVs可以逆转尼古丁引发的GC-1生精细胞增殖、迁移能力的损伤以及细胞凋亡。

Repairing effect of mouse bone marrow mesenchymal stem cell-derived extracellular vesicles on nicotine-induced damage of mouse spermatogenic cells

Objective: To construct an in vitro model of nicotine-induced injury of mouse spermatogenic cells and to explore the repairing effect of mouse bone marrow mesenchymal stem cell derived extracellular vesicles (mBMMSCs EVs) on the nicotine-induced damage of spermatogenic cells GC-1. Methods: Different concentrations (0, 8, 16, 32 μg/mL) of nicotine were used to treat spermatogenic cell GC-1 for 24, 48, 72 h, respectively; and the optimal concentration of nicotine and time of treatment to induce damage on GC-1 cells were screened by MTT assay. The spermatogenic cells GC-1 were divided into control group, nicotine group and nicotine+extracellular vesicle group. MTT, Transwell and apoptosis experiments were used to evaluate the changes in the biological characteristics of damaged GC-1 cell before and after the intervention of mBMMSCs EVs. Results: The results of MTT assay showed that the proliferation of GC-1 was significantly decreased when the concentration of nicotine was 32 μg/mL for 24 h (P<0.05), mBMMSCs-EVS increased cell proliferation after treatment with nicotine (P<0.05); compared with the control group, GC-1 cells in nicotine group showed significantly reduced migration and increased apoptosis(P<0.01); compared with the nicotine group, nicotine+extracellular vesicle group showed significantly increased cell proliferation and migration, and decreased the apoptotic ability(P<0.05 or<0.01). Conclusion: mBMMSCs-EVs may repair the damage of nicotine on the proliferation and migration of spermatogenic cells and inhibited nicotine-induced apoptosis of spermatogenic cells.