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目的通过检测不同剂量的直链烷基苯磺酸钠(LAS)对人张氏肝细胞(Chang liver cells)增殖、损伤、凋亡及坏死的影响,探讨LAS对人张氏肝细胞的毒性作用。方法体外培养人张氏肝细胞,将细胞分为对照组及LAS不同剂量组,分别为正常对照组(不含LAS的正常培养基培养细胞)、LAS低、中、高剂量组(37.5、75.0及150.0 mg/L)。各组细胞培养24 h后,倒置显微镜下观察细胞形态及结构;四甲基偶氮唑盐(MTT)法检测细胞生存率;检测丙二醛(MDA)、乳酸脱氢酶(LDH)、天门冬氨酸氨基转移酶(AST)水平以了解肝细胞损伤程度;Hoechst 33342及PI染色观察细胞凋亡及坏死情况。结果 LAS处理后细胞表面出现黑色颗粒,部分细胞变圆脱落并产生碎片,高剂量组出现细胞脱落、死亡。LAS各处理组细胞存活率明显低于对照组(P<0.01),MDA、LDH及AST水平也均明显高于对照组;细胞凋亡率与坏死率明显增加(P<0.01),且这些变化随LAS剂量增加更加明显(P<0.01)。结论 LAS可抑制人张氏肝细胞增殖,改变细胞形态,并导致细胞损伤、凋亡和坏死。
Objective To investigate the effects of LAS on the proliferation, injury, apoptosis and necrosis of human Chang liver cells by different doses of LAS, and to explore the toxic effects of LAS on human Chang’s hepatocytes . Methods Human Hepatocytes were cultured in vitro and divided into control and LAS different dose groups, which were normal control group (cultured in normal medium without LAS), LAS low, medium and high dose groups (37.5, 75.0 And 150.0 mg / L). Cell morphology and structure were observed under inverted microscope 24 hours after cell culture in each group. Cell viability was assayed by MTT assay. Malondialdehyde (MDA), lactate dehydrogenase (LDH), Tianmen Aspartate aminotransferase (AST) levels in order to understand the degree of liver cell damage; Hoechst 33342 and PI staining observed apoptosis and necrosis. Results After LAS treatment, black particles appeared on the cell surface, some of the cells turned round and fell into pieces, and the cells in the high dose group fell off and died. The cell viability in LAS treatment group was significantly lower than that in control group (P <0.01), and the levels of MDA, LDH and AST were also significantly higher than those in control group. The apoptosis rate and necrosis rate were significantly increased (P <0.01) With LAS dose increased more significantly (P <0.01). Conclusion LAS can inhibit the proliferation of human hepatocytes, change the cell morphology, and lead to cell injury, apoptosis and necrosis.