西罗莫司对肿瘤坏死因子α诱导的HepG2细胞脂质积聚的影响

发布时间：2018-07-04 19:16

本文选题：西罗莫司 + 非酒精性脂肪性肝炎　；参考：《中国临床药理学杂志》2017年06期

【摘要】：目的研究西罗莫司对肿瘤坏死因子α(TNF-α)诱导的HepG2细胞脂质积聚的影响。方法将棕榈酸(PA)负荷的HepG2细胞分为3组:对照组[0.2%牛血清白蛋白(BSA)+0.16 mmol·mL~(-1)PA],模型组(0.2%BSA+0.16 mmol·mL~(-1)PA+25 ng·mL~(-1)TNF-α),实验组(0.2%BSA+0.16mmol·mL~(-1)PA+25 ng·mL~(-1)TNF-α+10 ng·mL~(-1)西罗莫司)。各组处理24h后,以油红O染色观察HepG2细胞脂质蓄积情况;以蛋白质印迹法检测哺乳动物雷帕霉素靶蛋白(mTOR)及其下游核糖体蛋白P70S6激酶(P70S6K)总蛋白和磷酸化蛋白水平;以实时荧光定量聚合酶链式反应检测固醇调节原件结合蛋白1(SREBP1)、乙酰辅酶A羧化酶(ACC)、脂肪酸合成酶(FAS)mRNA的表达。结果在炎症状态下,西罗莫司可以明显抑制TNF-α诱导的HepG2细胞脂质积聚。同时,西罗莫司降低了TNF-α诱导的磷酸化mTOR(p-mTOR)及磷酸化P70S6K(p-P70S6K)蛋白表达:对照组、模型组及实验组的p-mTOR蛋白相对水平分别为1.00±0.20,3.11±0.60,2.38±0.50;这3组的p-P70S6K蛋白相对水平分别为1.00±0.30,3.67±0.60,1.62±0.50,模型组与对照组相比或者实验组与模型组相比,差异均有统计学意义(均P0.05)。西罗莫司还下调了SREBP1、ACC、FAS的mRNA表达:对照组、模型组及实验组的FAS mRNA相对水平分别是1.04±0.32,2.85±0.90,1.68±0.38;这3组的ACC mRNA相对水平分别是1.04±0.30,3.23±1.33,2.07±0.52;这3组的SREBP1的mRNA相对水平分别是1.01±0.16,3.85±1.30,2.82±0.57,模型组与对照组相比或者实验组与模型组相比,差异均有统计学意义(均P0.05)。结论西罗莫司通过抑制mTOR信号通路,减轻TNF-α诱导的HepG2细胞脂质积聚。
[Abstract]:Aim to study the effect of sirolimus on lipid accumulation in HepG2 cells induced by tumor necrosis factor 伪 (TNF- 伪). Methods HepG2 cells loaded with palmitic acid (PA) were divided into three groups: control group [0.2% bovine serum albumin (BSA) 0.16 mmol mL ~ (-1) PA], model group (0.2 BSA 0.16 mmol mL ~ (-1) PA 25 ng mL ~ (-1) TNF- 伪), experimental group (0.2 BSA 0.16mmol mL ~ (-1) PA 25 ng mL ~ (-1) sirolimus). After 24 hours of treatment, lipid accumulation in HepG2 cells was observed by oil red O staining, total protein and phosphorylated protein levels of rapamycin target protein (mTOR) and its downstream ribosomal protein P70S6 kinase (P70S6K) were detected by Western blot. The expression of steroid regulatory original binding protein 1 (SREBP1), acetyl-coA carboxylase (ACC) and fatty acid synthase (FAS) mRNA were detected by real-time fluorescence quantitative polymerase chain reaction. Results sirolimus could significantly inhibit TNF- 伪 induced lipid accumulation in HepG2 cells. Sirolimus also decreased the expression of phosphorylated mTOR (p-mTOR) and phosphorylated P70S6K (p-P70S6K) protein induced by TNF- 伪. The relative levels of p-mTOR protein in model group and experimental group were 1.00 卤0.20 卤3.11 卤0.602.38 卤0.50 and 1.00 卤0.30 卤0.601.67 卤0.601.62 卤0.50, respectively. There were significant differences between the model group and the control group or between the experimental group and the model group (P0.05). Sirolimus also down-regulated the expression of SREBP1ACC-FAS mRNA in the control group. The relative levels of FAS mRNA in model group and experimental group were 1.04 卤0.32 卤0.90 卤1.68 卤0.38, 1.04 卤0.30, 3.23 卤1.33 卤2.07 卤0.52, 1.01 卤0.163.85 卤1.303.85 卤1.30 卤2.82 卤0.57, respectively, and the relative levels of SREBP1 mRNA in model group and experimental group were 1.01 卤0.163.85 卤1.300.82 卤0.57, respectively, and the relative levels of ACC mRNA in these three groups were 1.04 卤0.30, 3.23 卤1.33, 2.07 卤0.52, 1.01 卤0.163.85 卤1.30, 2.82 卤0.57, respectively. The difference was statistically significant (P0.05). Conclusion sirolimus alleviates TNF- 伪 -induced lipid accumulation in HepG2 cells by inhibiting mTOR signaling pathway.
【作者单位】：重庆医科大学脂糖代谢性疾病重庆市重点实验室;
【基金】：国家自然科学基金资助项目(31640043) 重庆市第七批市科技计划基金资助项目(cstc2016jcyj A0545)
【分类号】：R96

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