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目的探索金属硫蛋白Ⅰ/Ⅱ(MetallothioneinⅠ/Ⅱ,MTⅠ/Ⅱ)对丙烯酰胺诱导星形胶质细胞损伤的保护作用。方法采用新生C57野生型(MT+/+)和MT敲除型(MT-/-)小鼠进行大脑皮层星形胶质细胞原代培养,并采用免疫细胞化学法鉴定细胞纯度。MT+/+和MT-/-两种细胞分别给予0.01、0.1和1 mmol/L丙烯酰胺处理,于不同时间点检测其细胞存活率,乳酸脱氢酶(LDH)漏出率和细胞凋亡率。结果细胞培养至15 d,两种细胞在形态学上无明显差异,细胞纯度达到90%以上。丙烯酰胺低剂量组(0.01 mmol/L)作用24 h后,MT-/-细胞存活率明显低于MT+/+,高剂量组(1 mmol/L)作用时间短于24 h,可见MT+/+细胞的存活率明显高于MT-/-,随着作用时间延长后,未见明显差异。丙烯酰胺作用24 h后,LDH漏出率和细胞凋亡率均随剂量依赖性增加,且丙烯酰胺中低剂量组(0.01和0.1 mmol/L)MT-/-细胞LDH漏出率和凋亡率明显高于MT+/+细胞。结论 MTⅠ/ⅡI对丙烯酰胺引起的星形胶质细胞损伤具有一定的保护作用。
Objective To explore the protective effect of metallothionein Ⅰ / Ⅱ (MTⅠ / Ⅱ) on the damage of astrocytes induced by acrylamide. Methods Primary cultured astrocytes were cultured with C57 wild type (MT + / +) and MT knockout (MT - / -) mice. The purity of the cells was assessed by immunocytochemistry. The cells were treated with 0.01,0.1 and 1 mmol / L acrylamide for MT + / + and MT - / - respectively. Cell viability, lactate dehydrogenase (LDH) leakage rate and apoptosis rate were detected at different time points. Results The cells were cultured for 15 days. There was no significant difference in morphology between the two cells. The purity of the cells reached more than 90%. The survival rate of MT - / - cells was significantly lower than that of MT + / + groups after treated with low dose acrylamide (0.01 mmol / L) for 24 h. The effect of MT + / + Cell survival rate was significantly higher than the MT - / -, with the role of prolonged time, no significant difference. After exposure to acrylamide for 24 h, LDH leakage rate and apoptosis rate increased in a dose-dependent manner, and LDH leakage rate and apoptosis rate of medium and low dose acrylamide groups (0.01 and 0.1 mmol / L) were significantly Higher than MT + / + cells. Conclusion MT Ⅰ / Ⅱ I can protect the astrocytes induced by acrylamide.