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目的观察食品添加剂亚硫酸钠对人肝肿瘤细胞(HepG2)的细胞毒性作用和脂肪变作用。方法向HepG2细胞悬液中分别加入终浓度为0(阴性对照)～10 mmol/L亚硫酸钠溶液染毒24、48和72 h,并设空白对照(无细胞)组和阳性对照组(含1%Tritonx-100的DMEM),每组6个复孔。采用乳酸脱氢酶(LDH)活力测定法检测肝细胞膜的通透性变化,采用CCK-8法检测肝细胞的活性改变,采用油红O染色法观察肝细胞内脂肪沉积情况。结果与阴性对照组相比,仅10 mmol/L亚硫酸钠染毒24 h后HepG2细胞培养上清中LDH活力的释放率增加(P<0.05);而各剂量亚硫酸钠染毒48和72 h后HepG2细胞培养上清中LDH活力的释放率无显著变化。与阴性对照组相比,各剂量亚硫酸钠染毒24、48、72 h后(除0.039 mmol/L染毒72 h外)HepG2细胞的存活率均显著下降(P<0.05),且细胞存活率随着亚硫酸钠染毒剂量的升高而降低。染毒24、48 h后,部分亚硫酸钠染毒组HepG2细胞内出现了极少量的脂滴,而阴性对照组却未发现;染毒72 h后,10 mmol/L亚硫酸钠染毒组出现明显脂滴。结论一定剂量的亚硫酸钠染毒可增加HepG2细胞膜的通透性,对肝细胞活性有明显的抑制作用,并且可引起肝细胞内甘油三酯的蓄积。
Objective To observe the cytotoxicity and steatosis of dietary sodium sulfite on HepG2 cells. Methods HepG2 cells were suspended in HepG2 cells at concentrations of 0 (negative control) ~ 10 mmol / L sodium sulfite for 24, 48 and 72 h, respectively. Blank control group (no cell) and positive control group (containing 1% Tritonx-100 in DMEM) with 6 replicate wells per group. The changes of permeability of hepatocyte membrane were detected by lactate dehydrogenase (LDH) activity assay. The activity of hepatocytes was detected by CCK-8 assay. The fat deposition in hepatocytes was observed by oil red O staining. Results Compared with the negative control group, the release rate of LDH activity in the supernatant of HepG2 cells increased only after 10 mmol / L sodium sulphite was applied for 24 h (P <0.05). However, the apoptotic rate of HepG2 cells increased after 48 and 72 h There was no significant change in the release rate of LDH activity in the culture supernatant. Compared with the negative control group, the survival rates of HepG2 cells decreased significantly (P <0.05) 24 h, 48 h and 72 h after exposure to various doses of sodium sulfite (except for 0.039 mmol / L for 72 h) Sodium sulfite dose to reduce the increase. After exposure to 24,48 h, a small amount of lipid droplets appeared in HepG2 cells in some sodium sulfite exposure groups, but not in the negative control group. After 72 h of exposure, significant lipid droplets appeared in the 10 mmol / L sodium sulfite exposure group . Conclusion A certain dose of sodium sulfite can increase the permeability of HepG2 cell membrane and significantly inhibit the activity of hepatocytes, and can induce the accumulation of triglycerides in hepatocytes.