生长分化因子11对甲醛诱导的海马神经细胞凋亡的影响

目的探讨生长分化因子11（GDF11）对甲醛诱导的海马神经（HT22）细胞毒性的影响。 方法把HT22细胞分为对照组（细胞未做任何处理）、甲醛组（50、100、200 μmol/ L甲醛处理细胞）和GDF11+甲醛组（GDF11转染细胞后用100 μmol/L甲醛处理）。细胞计数试剂盒（CCK8）法检测HT22细胞的活力；蛋白免疫印迹法检测HT22细胞凋亡相关蛋白Bax以及Bcl-2的变化；caspase-3活性检测试剂盒检测HT22细胞内caspase-3活性；DCFDA染色流式细胞仪检测HT22细胞中活性氧（ROS）水平。三组间比较采用单因素方差分析，组间两两比较采用LSD-t检验。 结果与对照组比较，甲醛组HT22细胞活力（92.23±0.20比56.12±0.61）和Bcl-2蛋白表达（220.32±2.21比150.25±0.31）水平均降低，差异具有统计学意义（P均< 0.05）；而caspase-3活性（95.36±1.74比190.17±2.14）、Bax蛋白表达（132.19±1.21比150.17±1.06）和ROS水平（1099.32±75.47比2802.17±126.49）均升高，差异具有统计学意义（P均< 0.05）。GDF11转染HT22细胞后，与甲醛组比较，GDF11+甲醛组HT22细胞活力升高（56.12±0.61比83.11±1.64），Bax蛋白表达（270.03±0.17比150.17±1.06）降低，Bcl-2蛋白表达（150.25±0.31比187.34±1.52）升高，caspase-3活性降低（190.17±2.14比105.31±4.12）和ROS水平降低（2802.17±126.49比1305.36±68.45），差异具有统计学意义（P均< 0.05）。 结论GDF11能够逆转甲醛对HT22细胞凋亡的诱导作用以及降低甲醛对HT22细胞ROS水平的增加作用，此机制对防治甲醛的神经毒性具有重要意义。

Effect of GDF11 on formaldehyde-induced apoptosis in HT22 cells

ObjectiveTo investigate effect of GDF11 on formaldehyde-induced cytotoxicity in HT22 cells. MethodsHT22 cells were divided into control group (cells without any treatment) , formaldehyde (FA) group (50, 100, 200 μmol/L formaldehyde respectively treated cells) and GDF11+FA group (GDF11 transfected cells and 100 μmol/L formaldehyde respectively treated cells) . Cell counting box (CCK8) was used to detect the viability of HT22 cells, and Western Blotting was used to detect the changes of apoptosis-related proteins Bax and Bcl-2 in HT22 cells. Caspase-3 activity in HT22 cells was detected by caspase-3 activity detection kit, and reactive oxygen species (ROS) level in HT22 cells was detected by flow cytometry (FCM) after DCFDA staining. One-way analysis was used to compare the differences among three groups. ResultsCompared with control group, the viability of HT22 cells (56.12±0.61 vs 92.23±0.20) and the expression level of Bcl-2 protein (220.32±2.21 vs 150.25±0.31) were decreased in the FA group (P < 0.05) . Caspase-3 activity (95.36±1.74 vs 190.17±2.14) , and the expression levels of Bax protein (132.19±1.21 vs 150.17±1.00) and ROS (1099.32±75.47 vs 2802.17±126.49) were significantly higher (P < 0.05) . After transfection of HT22 cells with GDF11, compared with FA group, the viability of HT22 cells in the GDF11+FA group was increased (56.12±0.61 vs 83.11±1.64) , the expression of Bax protein decreased (270.03±0.17 vs 150.17±1.06) , the expression of Bcl-2 protein increased (150.25±0.31 vs 187.34±1.52) , caspase-3 activity decreased (190.17±2.14 vs 105.31±4.12) , and the ROS level decreased (2802.17±126.49 vs 1305.36±68.45) . The difference was statistically significant (all P < 0.05) . ConclusionGDF11 can prevent formaldehyde-induced apoptosis and reduce ROS production in HT22 cells, which is of great significance in preventing and treating the neurotoxicity of formaldehyde.