低剂量氢醌诱导人胚肺成纤维细胞适应性反应研究

目的通过研究低剂量氢醌(HQ)诱导人胚肺成纤维细胞(HLF)适应性反应的量效关系,建立氧化适应模式,探讨真核细胞过氧化适应性反应及其可能机理。方法应用HLF,以HQ为诱导剂,用不同剂量HQ对HLF细胞预处理12h后,再用80μmol LHQ攻击HLF1h,结合微核试验、彗星实验及细胞周期变化,观察低剂量HQ诱导的适应性反应。结果在细胞整体活力水平,0.001μmol L、0.01μmol LHQ预处理可以提高HLF对攻击剂量80.0μmol L的耐受性。与对照组比较,从0.5μmol LHQ剂量开始预处理的HLF的微核率和核异常率明显增加(P<0.05),细胞出现不同程度拖尾现象,拖尾率及彗星尾长显著增加(P<0.01),从0.1μmol L预处理剂量开始,随着预处理剂量的增加,属3级、4级DNA损伤细胞比例逐渐增加。细胞周期出现G2期阻滞,G1期比例减少。与细胞仅大剂量攻击比较,0～0.1μmol L预处理的HLF微核率和核异常率、尾长及拖尾率均明显减少、DNA重度损伤比例明显下降(P<0.05)。相关分析表明,预处理剂量从0.1μmol L开始,HLF的微核率、核异常率、拖尾率及彗星尾长与HQ预处理剂量间均存在剂量反应关系。结论低剂量HQ预处理HLF后再用高剂量去攻击,出现不同程度的适应性反应。

Study on the adaptive reaction of low dose hydroquinone inducement in human embryo lung fibroblasts cells

OBJECTIVE: To study the adaptive reaction and its possible mechanisms in eukaryotic cells, the oxidize of adaptive mode was established by studying the dose-effect relation of inducing the adaptive reaction. METHODS: Human embryo lung fibroblasts (HLF) were attacked 1 hour by 80.0 micromol/L concentration respectively after using a different concentration HQ to pretreat the 12 hours. The adaptive reaction of a low concentration HQ inducement was observed by combining the micronucleus test, the comet test and the cell cycle change. RESULTS: The results showed that, in cellular viability, HLF induced by pretreated with HQ at 0.001 micromol/L and 0.0 micromol/L appeared to be tolerated when retreated by the following 80.0 micromol/L concentration of HQ. Compared with control respectively, the micronucleus rate and abnormal nuclei rate increased obviously by pretreatment with HQ concentration from 0.5 micromol/L to 80 micromol/L in HLF. Phenomena of cells with tails appeared, tail length and percentage of cells with tails increased significantly in HLF (P < 0.01). From 0.1 micromol/L pretreatment dose on, proportion of DNA lesion cells belonged to three and four grade increase gradually by the dose increasing. G2 phase arrest and cell numbers decreasing in G1. When compared with only high dose attack respectively, micronucleus rate, abnormal nuclei rate, tail length, percentage of cells with tails decreased significantly. proportion of severely damaged cells descended markedly by pretreatment with HQ from 0 micromol/L from 0.1 micromol/L in HLF (P < 0.05). It showed that there was a dose-related increase in the rate of micronucleus or abnormal nuclei, tail length and in percentage of cells with tails after pretreatment with HQ from 0 micromol/L to 80 micromol/L in HLF (P < 0.05). CONCLUSION: HLF induced by pretreated with HQ at low concentration appeared to adaptive reaction when retreated by high concentration of HQ.