收稿日期: 2017-07-12
网络出版日期: 2019-09-04
基金资助
国家自然科学基金资助项目(91543123);国家自然科学青年基金资助项目(21107068);创新团队发展计划资助项目(IRT13078)
Cytotoxicity in SH-SY5Y cells induced by $O_{3}$-oxidized black carbon particles
Received date: 2017-07-12
Online published: 2019-09-04
以人神经母细胞瘤细胞SH-SY5Y为模型, 在细胞体系探讨经O$_{3}$氧化过的黑碳(O$_3$-oxidized black carbon, OBC)颗粒对神经细胞的毒性作用. 应用MTT方法评估SH-SY5Y细胞的存活率; 通过测定培养液中乳酸脱氢酶(lactate dehydrogenase, LHD)浓度评估细胞膜的损伤程度; 利用流式细胞术评价细胞凋亡率; 使用荧光探针(Fluo 3-AM)检测细胞内Ca$^{2+}$浓度. 结果显示, OBC颗粒对SH-SY5Y细胞暴露24和48 h的半致死浓度(IC$_{50})$分别为202.2和133.3 $\mu $g/mL; 随着BC颗粒染毒浓度的增加(0, 5, 10, 20, 40 $\mu $g/mL), 胞外LDH浓度、细胞凋亡率和Ca$^{2+}$浓度均逐渐升高, 呈显著的剂量-效应关系($p<$0.05). 研究表明, OBC颗粒可导致神经细胞内Ca$^{2+}$浓度升高、 细胞膜损坏, 并最终导致细胞凋亡的发生.
尚羽, 王田田, 吴美英, 王璐, 何慧心, 安静 . $O_{3}$氧化黑碳对SH-SY5Y细胞的毒性作用[J]. 上海大学学报(自然科学版), 2019 , 25(4) : 550 -557 . DOI: 10.12066/j.issn.1007-2861.1963
SH-SY5Y cells were used to investigate neurotoxicity of O$_3$-oxidized black carbon (OBC) particles. Cell viability was measured by MTT assay after being treated with OBC for 24 and 48 h. Lactate dehydrogenase (LDH) leakage was determined to evaluate cellular membrane injury after OBC treatment for 24 h. Apoptosis rate was detected by cytometry after OBC treatment for 24 h. Concentrations of Ca$^{2+}$ was then measured with fluorescent probe of Fluo 3-AM. OBC exposure caused significant dose-dependent and time-dependent viability decrease. LC$_{50}$ for 24 and 48 h were 202.2 and 133.3 $\mu $g/mL, respectively. The LDH leakage, apoptosis rate and intracellular Ca$^{2+}$ levels were also increased significantly in a dose-dependent manner after being treated with OBC in SH-SY5Y cells. These results suggest that OBC may mediate cytotoxic effects and apoptosis through Ca$^{2+}$ release.
Key words： black carbon (BC) particles; SH-SY5Y cells; cytotoxicity; apoptosis; Ca$^{2+}$
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