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目的探讨亚硫酸钠(Na2SO3)对人胚肾293(HEK293)细胞炎性因子表达的影响。方法采用改进的单溶液噻唑兰(MTS)比色法观察0、0.0025、0.01、0.039、0.156、0.625、2.5、10mmol/L亚硫酸钠对人胚肾293细胞株(HEK293)的毒性作用,并观察0、0.039、0.156、0.625、2.5、10mmol/L亚硫酸钠染毒致HEK293细胞的形态学改变,采用逆转录-聚合酶链反应(RT-PCR)法检测HEK293细胞中肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)和白介素8(IL-8)的mRNA水平的改变。结果细胞毒性实验显示,当亚硫酸钠浓度为0.625、2.5、10mmol/L时,OD值分别为(0.35475±0.02124),(0.60050±0.01277),(0.78475±0.00985),均低于对照组(2.5145±0.202265),差异有统计学意义(P<0.05);当亚硫酸钠浓度≤0.156mmol/L时,OD值范围为(2.47375±0.06999)～(2.62500±0.12029),与对照组比较,差异无统计学意义。形态学观察发现,当接触亚硫酸钠浓度≥0.625mmol/L时,细胞数量明显降低,细胞比较细长,突起较少;当亚硫酸钠浓度≤0.156mmol/L时,细胞数量和形态基本无变化。RT-PCR检测结果显示,0.039～10mmol/L的亚硫酸钠对TNF-α、MCP-1和IL-8的mRNA表达无明显诱导作用。结论亚硫酸钠对人胚肾细胞具有一定的抑制损伤作用,但对TNF-α、MCP-1和IL-8基因表达水平无影响,二者之间无明显关联。
Objective To investigate the effect of sodium sulfite (Na2SO3) on the expression of inflammatory cytokines in human embryonic kidney 293 (HEK293) cells. Methods The toxic effects of sodium sulfite (0,0.0025,0.01,0.039,0.156,0.625,2.5,10mmol / L) on human embryonic kidney 293 cell line (HEK293) were observed by the modified single-solvent thiazolyl (MTS) , 0.039,0.156,0.625,2.5,10 mmol / L sodium sulfite. HEK293 cells were treated with TNF-α by reverse transcription-polymerase chain reaction (RT-PCR) , Monocyte chemotactic protein 1 (MCP-1) and interleukin 8 (IL-8). Results Cytotoxicity experiments showed that when the concentration of sodium sulfite was 0.625, 2.5 and 10 mmol / L, the OD values were (0.35475 ± 0.02124) and (0.60050 ± 0.01277) and (0.78475 ± 0.00985), respectively, which were lower than that of the control group (2.5145 ± 0.202265 ), The difference was statistically significant (P <0.05). When the concentration of sodium sulfite was0.156mmol / L, the OD value ranged from (2.47375 ± 0.06999) to (2.62500 ± 0.12029), there was no significant difference compared with the control group. Morphological observation showed that when the concentration of sodium sulfite was ≥0.625mmol / L, the number of cells was significantly decreased, the cells were slender and the number of processes was less. When the concentration of sodium sulfite was less than or equal to0.156mmol / L, the number and shape of cells were almost unchanged. The results of RT-PCR showed that 0.039 ~ 10mmol / L sodium sulfite had no obvious effect on the mRNA expression of TNF-α, MCP-1 and IL-8. Conclusion Sodium sulfite has a certain inhibitory effect on human embryonic kidney cells, but has no effect on the expression of TNF-α, MCP-1 and IL-8. There is no significant correlation between them.