目的探讨竹荪多糖对亚砷酸钠致人胎肝(L-02)细胞毒性的拮抗作用。方法将处于对数生长期的L-02细胞分别暴露于含不同浓度[0(对照)~80μmol/L]亚砷酸钠和[0(对照)~160μg/ml]竹荪多糖+10μmol/L亚砷酸钠的培养液中暴露24 h,采用四甲基偶氮唑蓝(MTT)法测定细胞存活率。将处于对数生长期的L-02细胞分设对照(培养基)组、亚砷酸钠(10μmol/L)组、竹荪多糖(80μg/ml)组和联合处理(10μmol/L亚砷酸钠+80μg/ml竹荪多糖)组,暴露24 h后,检测细胞培养液中丙二醛(MDA)、总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、还原性谷胱甘肽(GSH)、细胞色素C(Cyt-C)水平和天冬氨酸蛋白水解酶3(Caspase-3)的活力及B淋巴细胞瘤-2(Bcl-2)蛋白、Bcl-2相关X(Bax)蛋白的表达水平。结果与对照组比较,除2.5和5μmol/L亚砷酸钠染毒组外,10~80μmol/L亚砷酸钠染毒组L-02细胞的存活率均下降,差异有统计学意义(P<0.05);且随着亚砷酸钠染毒剂量的升高,L-02细胞的存活率呈下降趋势。与对照组比较,40、80和160μg/ml竹荪多糖联合处理组L-02细胞的存活率均升高,差异有统计学意义(P<0.05);其中,以80μg/ml竹荪多糖的拮抗效果最好。与对照组比较,亚砷酸钠组和联合处理组L-02细胞培养液中MDA和Cyt-C的水平升高,T-SOD、CAT及GSH的水平降低,差异均有统计学意义(P<0.05);而竹荪多糖组L-02细胞培养液中上述指标均无明显变化。与亚砷酸钠组比较,联合处理组和竹荪多糖组L-02细胞培养液中MDA和Cyt-C的水平降低,T-SOD、CAT及GSH的水平升高,差异均有统计学意义(P<0.05)。与对照组比较,亚砷酸钠组和联合处理组L-02细胞胞内Caspase-3的活力升高,差异具有统计学意义(P<0.05);而竹荪多糖组L-02细胞细胞内Caspase-3的活力无明显变化。与亚砷酸钠组比较,联合处理组和竹荪多糖组L-02细胞内Caspase-3的活力均降低,差异有统计学意义(P<0.05)。与对照组比较,亚砷酸钠组和联合处理组L-02细胞胞内Bcl-2、Bax蛋白的表达水平升高,Bcl-2/Bax比值降低,差异均有统计学意义(P<0.05);而竹荪多糖组L-02细胞上述指标均无明显变化。与亚砷酸钠组比较,联合处理组和竹荪多糖组L-02细胞胞内Bax蛋白的表达水平降低,Bcl-2/Bax比值升高;竹荪多糖组L-02细胞内Bcl-2蛋白的表达水平降低,差异均有统计学意义(P<0.05)。结论亚砷酸钠可以诱导L-02细胞氧化应激,通过激活线粒体途径的细胞凋亡发挥毒性作用;竹荪多糖可以通过抑制上述机制,拮抗亚砷酸钠所致L-02细胞的毒性作用。 Objective To investigate the antagonistic effect of Dictyophragus monocytogenes on cytotoxicity induced by sodium arsenite in human fetal liver (L-02). Methods L-02 cells in logarithmic growth phase were exposed to different concentrations of [0 (control) ~80μmol / L] sodium arsenite and [0 (control) ~ 160μg / ml] Sodium arsenite culture medium exposure 24 h, the use of tetramethyl blue (MTT) assay cell viability. L-02 cells in the logarithmic growth phase were divided into control (medium) group, sodium arsenite (10μmol / L) group, polysaccharides (80μg / ml) and combined treatment +80 μg / ml Dictyostelium polysaccharide). After being exposed for 24 h, the contents of malondialdehyde (MDA), total superoxide dismutase (T-SOD), catalase (CAT) The levels of GSH, Cyt-C and Caspase-3 and Bcl-2, Bcl-2 X (Bax) protein expression levels. Results Compared with the control group, the survival rates of L-02 cells treated with 10 ~ 80μmol / L sodium arsenite were significantly decreased except 2.5 and 5μmol / L sodium arsenite exposure group (P <0.05). And with the increase of sodium arsenite dose, the survival rate of L-02 cells showed a decreasing trend. Compared with the control group, the survival rates of L-02 cells in 40, 80 and 160μg / ml Dictyostelium polysaccharide combined treatment group were significantly increased (P <0.05), among which 80μg / ml Dictyostelium The best antagonistic effect. Compared with the control group, the levels of MDA and Cyt-C, the levels of T-SOD, CAT and GSH in L-02 cell culture fluid in sodium arsenite group and combined treatment group were decreased, the differences were statistically significant (P <0.05). However, no significant changes were observed in the L-02 cell culture medium of Dictyostelium polysaccharide group. Compared with the sodium arsenite group, the levels of MDA and Cyt-C, the levels of T-SOD, CAT and GSH in the L-02 cell culture medium of the combined treatment group and the polysaccharide-extract group were significantly increased (P <0.05). Compared with the control group, the intracellular Caspase-3 activity of L-02 cells in the sodium arsenite group and the combined treatment group was significantly increased (P <0.05), while that in the L-02 cells No significant changes in the vitality of Caspase-3. Compared with sodium arsenite group, the activity of Caspase-3 in L-02 cells of combination treatment group and Dictyostelium polysaccharide group decreased, the difference was statistically significant (P <0.05). Compared with the control group, the expression of Bcl-2 and Bax protein and the ratio of Bcl-2 / Bax in L-02 cells in sodium arsenite group and combination group decreased significantly, with statistical significance (P <0.05 ); And Dictyostelium polysaccharide L-02 cells showed no significant changes in the above indicators. Compared with sodium arsenite group, the expression of Bax protein and the ratio of Bcl-2 / Bax in L-02 cells in combination treatment group and Dictyostelium polysaccharide group were significantly decreased, and the Bcl-2 The protein expression decreased, the differences were statistically significant (P <0.05). Conclusion Sodium arsenite can induce oxidative stress in L-02 cells and exert toxic effects by activating mitochondrial pathway of apoptosis. Dictyostelium polysaccharide can inhibit the toxic effects of sodium arsenite-induced L-02 cells by inhibiting the above mechanism .