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目的探讨金雀异黄素(genistein,GEN)对L-02人肝细胞DNA辐射损伤的防护作用。方法 (1)L-02细胞经不同浓度(1、5、10、20μmol/L)GEN预处理24 h后,接受不同剂量(6、8、12、16、20 Gy)X线照射,利用MTT法检测细胞增殖情况。(2)L-02细胞经不同浓度GEN预处理后接受8 Gy(300 cGy/min)X线照射,利用单细胞凝胶电泳检测照射后不同时间(24、48、72 h)细胞DNA损伤情况。另设正常对照(N)、单纯GEN处理(G)及单纯照射(R)组作为参照。结果 (1)当照射剂量为6、8及12 Gy时,5μmol/L GEN预处理组细胞增殖率显著高于R组(P<0.05),而照射剂量为16 Gy和20 Gy时,各浓度GEN预处理组细胞增殖率与R组相比未见明显升高。(2)DNA损伤检测结果显示,N组和不接受照射的各浓度G组细胞均未见彗星拖尾。经8 Gy X线照射后24 h,R组及各浓度GEN预处理组彗星出现率均小于1%,各组间彗星尾长差异无统计学意义;照后48 h,R组彗星出现率达(24.2±1.2)%,彗星尾长达(283.6±22.3)μm,而各浓度GEN预处理组的彗星出现率和彗星尾长均较R组明显降低(P<0.05);照后72 h,R组彗星出现率较48 h明显下降(P<0.05)、彗星尾长明显缩短(P<0.05),1、5μmol/L GEN预处理组彗星出现率和彗星尾长仍明显低于R组(P<0.05),但10、20μmol/L GEN预处理组彗星出现率则明显升高(P<0.05),彗星尾长明显增长(P<0.05)。结论低浓度(1、5μmol/L)GEN能有效减轻较低剂量(8Gy)照射对L-02人肝细胞的DNA辐射损伤。
Objective To investigate the protective effect of genistein (GEN) on DNA damage induced by L-02 human hepatocytes. Methods (1) L-02 cells were pretreated with different concentrations of GEN (1,5, 10 and 20μmol / L) for 24 hours and then received different doses of 6,8,12,16,20 Gy X-ray. MTT Method to detect cell proliferation. (2) X-ray irradiation of 8 Gy (300 cGy / min) was performed on L-02 cells pretreated with different concentrations of GEN. DNA damage was detected by single cell gel electrophoresis at different times (24, 48 and 72 h) . Another set of normal control (N), simple GEN treatment (G) and simple irradiation (R) group as a reference. Results (1) When the irradiation doses were 6, 8 and 12 Gy, the cell proliferation rate in 5 μmol / L GEN pretreatment group was significantly higher than that in R group (P <0.05). At the irradiation dose of 16 Gy and 20 Gy, The cell proliferation rate in GEN pretreatment group was not significantly higher than that in R group. (2) The result of DNA damage test showed that there were no comet tailing in N and non-irradiated G groups. At 24 h after irradiation with 8 Gy X-ray, the incidence of comets in R group and each concentration of GEN pretreatment group was less than 1%, and there was no significant difference in the tail length of comet between groups. At 48 h after irradiation, the incidence of comet in R group reached (24.2 ± 1.2)% and comet tail (283.6 ± 22.3) μm, respectively. The comet and tail lengths of comets in each concentration of GEN preconditioning group were significantly lower than those in R group (P <0.05) The incidence of comet in R group was significantly lower than that in 48 h (P <0.05), and the tail length of comet was significantly shortened (P <0.05). The incidence of comet and tail of comet in 1,5 μmol / L GEN pretreatment group were still significantly lower than those in R group (P <0.05). However, the incidences of comets in 10 and 20μmol / L GEN pretreatment groups were significantly increased (P <0.05), and the tail length of comet significantly increased (P <0.05). Conclusion GEN (1μmol / L) at low concentration can effectively attenuate the DNA damage induced by low dose (8Gy) radiation on L-02 human hepatocytes.