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目的检测超氧化物歧化酶(SOD)活性等抗氧化指标在青蒿琥酯诱导K562细胞凋亡过程中的变化,探讨青蒿琥酯诱导K562细胞凋亡过程中的氧化损伤机制。方法不同浓度(1×10-3、2×10-4、4×10-5mol/L)的青蒿琥酯作用下培养K562细胞,同时以未加青蒿琥酯的K562细胞作为对照组,采用Annexin V-PI双染和Hochest 33258、黄嘌呤氧化酶法和实时荧光定量PCR法分别检测凋亡、SOD活性和锰超氧化物歧化酶(MnSOD)、含硒谷胱甘肽过氧化物酶(SeGPx)mRNA的表达情况。结果 SOD活性:4×10-5mol/L和2×10-4mol/L两浓度组与对照组比较差异有统计学意义(P<0.05),浓度为4×10-5mol/L、2×10-4mol/L组与1×10-3mol/L组比较差异有统计学意义(P<0.05),1×10-3mol/L组与对照组比较差异无统计学意义(P>0.05),4×10-5mol/L组与2×10-4mol/L组比较差异无统计学意义(P>0.05)。SeGPx及MnSOD mRNA表达:4×10-5mol/L、2×10-4mol/L两浓度组与对照组比较差异有统计学意义(P<0.05),4×10-5mol/L、2×10-4mol/L组与1×10-3mol/L组比较差异有统计学意义(P<0.05),1×10-3mol/L组与对照组比较差异无统计学意义(P>0.05),4×10-5mol/L组与2×10-4mol/L组比较差异无统计学意义(P>0.05)。各浓度组中,以4×10-5mol/L组的SeGPx mRNA、MnSOD mRNA表达最高。结论氧化损伤机制参与了青蒿琥酯诱导K562细胞凋亡的过程,但氧化损伤可能不是青蒿琥酯诱导K562细胞凋亡的主要机制。
Objective To detect the changes of antioxidant enzymes such as superoxide dismutase (SOD) activity during the apoptosis of K562 cells induced by artesunate, and to explore the mechanism of oxidative damage induced by artesunate in K562 cells. Methods K562 cells were cultured under different doses of artesunate (1 × 10-3, 2 × 10-4 and 4 × 10-5 mol / L). K562 cells without artesunate were used as control group, Annexin V-PI double staining and Hochest 33258, xanthine oxidase and real-time fluorescence quantitative PCR were used to detect apoptosis, SOD activity and manganese superoxide dismutase (MnSOD), selenium-containing glutathione peroxidase (SeGPx) mRNA expression. Results SOD activity was significantly different between the two concentrations of 4 × 10-5mol / L and 2 × 10-4mol / L compared with the control group (P <0.05), the concentration of 4 × 10-5mol / L, 2 × 10 (P <0.05). There was no significant difference between 1 mol / L group and 1 × 10-3 mol / L group (P> 0.05) There was no significant difference between × 10-5mol / L group and 2 × 10-4mol / L group (P> 0.05). The expressions of SeGPx and MnSOD mRNA were significantly different between 4 × 10-5mol / L and 2 × 10-4mol / L groups (P <0.05), 4 × 10-5mol / L and 2 × 10 (P <0.05). There was no significant difference between 1 mol / L group and 1 × 10-3 mol / L group (P> 0.05) There was no significant difference between × 10-5mol / L group and 2 × 10-4mol / L group (P> 0.05). In each concentration group, the expression of SeGPx mRNA and MnSOD mRNA was the highest in 4 × 10-5mol / L group. Conclusion The mechanism of oxidative damage is involved in artesunate-induced K562 cell apoptosis, but oxidative damage may not be the main mechanism of artesunate-induced apoptosis in K562 cells.