几丁聚糖对氯化镉诱导HepG2细胞产生ROS水平的影响

目的研究不同浓度几丁聚糖对氯化镉诱导HepG2细胞产生活性氧(ROS)的清除作用。方法实验分设6组:正常对照组(仅含10%新生牛血清的培养液,无氯化镉和几丁聚糖)、20μmol/L氯化镉组、0.50mg/ml几丁聚糖组、20μmol/L氯化镉+几丁聚糖三个联合作用组(浓度分别为0.02、0.10和0.50mg/ml)。HepG2细胞处理时间为4h。以氯化镉诱导HepG2细胞产生ROS,运用DCFH-DA作为荧光探针,采用流式细胞检测技术检测几丁聚糖与氯化镉联合作用下HepG2细胞内ROS水平。结果20μmol/L氯化镉可使HepG2细胞内ROS水平显著增加(P<0.01);0.50mg/ml几丁聚糖处理HepG2细胞后,细胞内ROS水平较正常对照组略有降低,但并无显著性差异;与氯化镉染毒组相比,几丁聚糖和氯化镉联合作用组细胞内ROS水平随几丁聚糖浓度的增加,其DCF密度值由275.593逐渐降低到174.597,并且在中、高剂量几丁聚糖(0.1mg/ml和0.5mg/ml)和氯化镉联合作用组差异有极显著性(P<0.01)。结论几丁聚糖对HepG2细胞内由氯化镉诱导而产生的ROS水平具有明确的调节作用。

Effect of chitosan on the level of ROS in HepG2 cell induced by cadmium chloride

OBJECTIVE: To study the effect of chitosan on ROS levels in HepG2 cell induced by cadmium chloride. METHODS: The experiments were divided into 6 groups: normal control group, cadmium chloride group (20micromol/L), chitosan group (0.50mg/ml) and three combined treatment group (cadmium chloride 20micromol/L and chitosan 0.02mg/ml, 0.10mg/ml and 0.50mg/ml). The HepG2 cells were treated by chitosan and cadmium chloride simultaneously for 4h. The fluorescence of 2', 7'-dichlorofluorescin (DCF) was measured by floweytometry as a mean of estimating the formation of reactive oxygen species (ROS). RESULTS: In the comparison with normal control, the DCF intensity of chitosan group only exhibited somewhat decrease (P > 0.05) and significantly increased in cadmium chloride group (20micromol/L) (P < 0.01). The DCF intensity decreased in all three combined groups were more higher than those of cadmium chloride group, and when the concentration of chitosan improved from 0.01mg/ml to 0.50mg/ml, the DCF intensity decreased from 275.593 to 174.597 and the DCF intensity significant decrease was observed at concentration of 0.10mg/ml and 0.50mg/ml chitosan. CONCLUSION: It was suggested that chitosan could antagonize the increase of ROS in HepG2 cell induced by cadmium chloride.