目的研究不同剂量的锰(Mn)对PC12细胞生长增殖的影响。方法体外培养大鼠嗜铬细胞瘤PC12细胞。以100,900μmol/LMnCl2染毒后进行四唑盐比色实验(MTT)观察MnCl2对细胞的抑制作用,TUNEL法观察细胞凋亡,Western blot检测α-共核蛋白(α-SYN)表达情况。结果MTT结果显示100—900μmol/L MnCl2对细胞的生长增殖有显著的抑制作用(P〈0.01)。TUNEL染色结果显示MnCl2处理组TUNEL阳性细胞增多(P〈0.05)。Westem blot结果显示与对照组相比,100、300、500μmol/L MnCl2组α-SYN表达水平分别升高1.8、4.5、7.3倍(P〈0.05)。结论一定剂量的MnCl2对PCl2细胞的生长增殖有明显的抑制作用,这一作用可能是通过诱导胞内α-SYN表达增高实现的。提示预防、阻止α-SYN在细胞中的聚集可能会降低Mn对多巴胺能神经细胞的损伤。
Objective To study the toxicity of different doses of manganese chloride (MnCl2) on rat pheochromocytoma PC12 cells in vitro.Methods PC12 cell cultures were exposed to MnC12 within a dose range of 100-900 μmol/L. The inhibition of the cells during 7 days was determined by MTT assay, the apoptotic cells were monitored by TUNEL stain. The alpha-synuclein were determined by western blot. Results MnCl2 at different concentrations (100, 300, 500, 700, 900 μmol/L) could inhibit the growth and proliferation of PC12 cells in a dose and time-dependent manner and the cell growth curve was changed accordingly. After the treatment of 300,500 μmol/L MnCl2 ,TUNEL-positive cells were also increased (P 〈 0.05). Westernblot showed that MnCl2 increased the alphasynuclein in a dose-dependent manner ( P 〈 0.05). Conclusion MnCl2 can significantly inhibit the growth and proliferation of PC12 cells, possibly by inducing apeptosis,and the increase of alpha-synuclein.