2021 年第 1 期 第 16 卷

丙酮醛对体外施万细胞的损伤作用研究

Injurious effects of methylglyoxal on Schwann cells in vitro

作者：陈露露祁佐良

英文作者：Chen Lulu Qi Zuoliang

单位：中国医学科学院北京协和医学院整形外科医院整形16科，北京100144

英文单位：The 16th Department of Plastic Surgery Hospital Peking Union Medical College Chinese Academy of Medical Sciences Beijing 100144 China

关键词：施万细胞；丙酮醛；细胞活性；细胞凋亡

英文关键词：Schwanncells；Methylglyoxal;Cellactivity;Cellapoptosis

• 摘要：

• 目的 探讨不同浓度丙酮醛对体外施万细胞的损伤作用。方法将RSC96施万细胞体外培养48 h后，改用不含血清的培养液培养12 h。实验分为空白对照组和1.0、1.2、1.4、1.6、1.8、2.0 mmol/L丙酮醛组。空白对照组不加任何药物处理，其余各组分别加入对应浓度丙酮醛，分别继续培养6 h和24 h后，显微镜下观察细胞形态变化。应用细胞计数盒8（CCK-8）试剂盒检测不同浓度丙酮醛对RSC96施万细胞活性的影响，应用Muse细胞检测分析仪检测各组细胞存活和凋亡情况。结果 RSC96施万细胞培养6、24 h后显微镜下观察发现，与无损伤的空白对照组相比，1.0 mmol/L丙酮醛组细胞形态均无明显变化；1.6 mmol/L丙酮醛组细胞形态均出现明显异常，显微镜下可见明显的细胞膜出芽现象，细胞质内出现大量的黑色颗粒物，胞核可见明显的凝集浓缩；随着丙酮醛剂量的增加施万细胞的损伤程度加重。1.0、1.2、1.4、1.6、1.8、2.0 mmol/L丙酮醛组RSC96施万细胞培养6、24 h后细胞活性和细胞存活率均低于空白对照组，细胞凋亡率均高于空白对照组［培养6 h：（5.0±0.9）%、（5.9±0.6）%、（6.7±0.3）%、（6.9±0.9）%、（8.6±0.5）%、（9.2±0.9）%比（3.8±0.6）%，培养24 h：（17.4±1.7）%、（25.7±3.6）%、（31.4±3.4）%、（36.9±2.4）%、（42.4±3.1）%、（44.2±5.2）%比（10.6±1.8）%］，且细胞存活率和细胞凋亡率均与丙酮醛浓度呈剂量依赖性（均P＜0.05）。培养24 h后不同浓度丙酮醛组RSC96施万细胞的存活率均低于本组培养6 h，凋亡率均高于本组培养6 h（均P＜0.05）。结论 体外丙酮醛对施万细胞具有明显的毒性作用，并可最终导致细胞凋亡。

• Objective To explore the injurious effects of methylglyoxal (MGO) with different concentrations on Schwann cells in vitro. Methods RSC96 Schwann cells were cultured in vitro for 48 h， and then cultured in culture medium with serum-free for 12 h. The Schwann cells were divided into blank control group, and 1.0, 1.2, 1.4, 1.6, 1.8 and 2.0 mmol/L MGO groups. The blank control group was not treated with any drug, and other groups were added with corresponding concentrations of MGO, respectively. After 6 and 24 h of continue culture, the morphological changes of cells were observed under microscope. Cell counting kit-8 (CCK-8) was used to analyze the effect of MGO in different concentrations on RSC96 Schwann cells activities. Muse cell detection analyzer was used to detect survival and apoptosis of cells. Results After 6 and 24 h culture, RSC Schwann cells were observed under microscope. Compared with control group, there were no significant changes of cell morphology in 1.0 mmol/L MGO group. The cell morphology of 1.6 mmol/L MGO group were significantly abnormal. The cell membranes with sprouting phenomenon, a large number of black particles in cytoplasm， and agglutination and condensation of nucleus were significantly observed under the microscope. The injurious degree of Schwann cells was aggravated with the increasing concentration of MGO. After 6 and 24 h of culture, the activities and survival rates of RSC96 Schwann cells in 1.0, 1.2, 1.4, 1.6, 1.8 and 2.0 mmol/L MGO groups were lower than those in blank control group, apoptosis rates in different concentrations groups were higher than those in blank control group ［6 h after culture：（5.0±0.9）%, （5.9±0.6）%, （6.7±0.3）%, （6.9±0.9）%, （8.6±0.5）%, （9.2±0.9）% vs （3.8±0.6）%; 24 h after  culture：（17.4±1.7）%, （25.7±3.6）%, （31.4±3.4）%, （36.9±2.4）%, （42.4±3.1）%, （44.2±5.2）% vs （10.6±1.8）%］, and the cells survival rate and apoptosis rate were dose-dependent with the concentration of MGO (all P<0.05). After 24 h of culture, the survival rate of RSC96 Schwann cells in different concentrations MGO groups were lower and the apoptosis were higher than those 6 h after culture (all P<0.05). Conclusion In vitro, MGO has obvious toxic effects on Schwann cells and lead to Schwann cells apoptosis eventually.