鱼藤酮对小鼠小胶质细胞 BV-2存活率及一氧化氮含量的影响

Effects of rotenone on survival rate and nitric oxide levels of mouse microglia cell line BV-2

目的：研究鱼藤酮对小鼠小胶质细胞BV-2存活率及一氧化氮含量的影响。方法 BV-2细胞以5×10^7/L密度接种到细胞培养板中，分别加入鱼藤酮0、1×10^-11、1×10^-10、1×10^-9、1×10^-8、1×10^-7、1×10^-6、1×10^-5 mol/L后0、6、12、24、48、72 h等测定细胞存活率。另以1×10^-8 mol/L鱼藤酮干预BV-2细胞48 h，测定上清液中超氧化物歧化酶、过氧化物酶、总巯基、超氧阴离子和NO含量并与未给予鱼藤酮干预的对照组比较。结果5×10^7/L BV-2细胞于0、6、12、24、48、72 h细胞增殖活力分别为0．035±0．001、0．132±0．006、0．334±0．017、1．073±0．044、2．272±0．172、0．776±0．032，可见48 h达到细胞生长曲线的峰值。鱼藤酮（1×10^-6 mol/L ）干预 BV-2细胞24、48、72 h 细胞存活率分别降低至（63．4±10．1）％、（51．7±12．2）％、（33．9±11．2）％；鱼藤酮（1×10^-7 mol/L）干预BV-2细胞72 h细胞存活率降低至（50．8±2．9）％。1×10^-8 mol/L 鱼藤酮干预48 h 后 BV-2细胞上清液一氧化氮水平达（27．6±6．2）μmol/L，明显高于对照组的（13．3±2．5）μmol/L （t＝-2．135， P＝0．044）。结论鱼藤酮在一定浓度范围内可激活小胶质细胞，但是超出此浓度范围时则可能导致小胶质细胞存活率降低。

Objective To investigate the effects of rotenone on survival rate and nitric oxide （ NO） levels of mice microglia cell line BV-2.Methods 5 ×10^7 cells/L of BV-2 cells were inoculated in cell culture plate and were treated by 1 ×10^-11 , 1 ×10^-10 , 1 ×10^-9 , 1 ×10^-8 , 1 ×10^-7 , 1 ×10^-6 or 1 ×10^-5 mol/L of rotenone; survival rates were analyzed at the time points of 0, 6, 12, 24, 48 and 72 hours.Superoxide dismutase （SOD）, peroxi-dase （ POD） and levels of total hydrosulphonyl groups （-SH） , superoxide anion （ O2-） and NO were measured 48 hours after treatment with 1 ×10^-8 mol/L rotenone.Results Cell viability of BV-2 was 0.035 ±0.001, 0.132 ± 0.006, 0.334 ±0.017, 1.073 ±0.044, 2.272 ±0.172 and 0.776 ±0.032 after rotenone treatment at 0, 6, 12, 24, 48 and 72 hours respectively.BV-2 cells reached to peak level of growth curve at 48 hours.Cell viability of BV-2 cells decreased to （63.4 ±10.1）%, （51.7 ±12.2）%and （33.9 ±11.2）%after treatment with rotenone （1 ×10^-6 mol/L） for 24, 48, 72 hours respectively.Cell viability of BV-2 was decreased to （50.8 ±2.9）%after rotenone （1 ×10^-7 mol/L） treatment for 72 hours.Supernatant NO level increased to （27.6 ±6.2） μmol/L in the group treated with 1 ×10^-8 mol/L rotenone for 48 hours and it was （13.3 ±2.5）μmol/L in the control group （t=-2.135, P=0.044）.Conclusion Rotenone activates microglial cells in certain range of concentrations but de-creases cell survival rate beyond the range .