TNF-α促进HepG2肝细胞脂质积聚及其机制的初步研究

发布时间：2018-04-27 08:07

  本文选题：TNF-α + 软脂酸　； 参考：《第三军医大学学报》2013年11期

【摘要】：目的探讨肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)是否能够促进肝细胞脂质积聚,并对其机制进行初步探讨。方法将HepG2肝细胞分为空白对照组、单纯TNF-α组(TNF-α2ng/mL或20ng/mL)、软脂酸组(软脂酸0.08mmol/L或0.2mmol/L)及联合组(TNF-α2ng/mL联合软脂酸0.08mmol/L、TNF-α2ng/mL联合软脂酸0.2mmol/L、TNF-α20ng/mL联合软脂酸0.08mmol/L、TNF-α20ng/mL联合软脂酸0.2mmol/L),处理24h,应用化学酶促-比色法定量检测细胞内TG含量。进一步选取TNF-α20ng/mL和软脂酸0.08mmol/L,通过油红O染色观察HepG2细胞内脂质积聚情况;实时荧光定量PCR和Western blot检测HepG2细胞SREBP-1、FAS、ACCα的表达水平。结果①单纯TNF-α组TG含量[TNF-α2ng/mL组(0.344±0.093)μg/μg、TNF-α20ng/mL组(0.329±0.068)μg/μg]分别较空白对照组[(0.192±0.048)μg/μg]显著升高(P0.05);联合组[TNF-α2ng/mL联合软脂酸0.08mmol/L组(0.451±0.096)μg/μg、TNF-α2ng/mL联合软脂酸0.2mmol/L组(0.821±0.257)μg/μg、TNF-α20ng/mL联合软脂酸0.08mmol/L组(1.032±0.286)μg/μg、TNF-α20ng/mL联合软脂酸0.2mmol/L组(2.134±1.049)μg/μg]分别较软脂酸组[软脂酸0.08mmol/L组(0.247±0.069)μg/μg、软脂酸0.2mmol/L组(0.341±0.031)μg/μg]显著升高(P0.05);②油红O染色进一步显示,TNF-α促进肝细胞内脂质积聚。③实时荧光定量PCR和Western blot检测结果显示单纯TNF-α组与空白对照组相比,HepG2细胞SREBP-1、FAS、ACCα的表达均增加(P0.05);联合组与软脂酸组相比,肝细胞内SREBP-1、FAS、ACCα的表达水平明显上调(P0.05)。结论TNF-α促进HepG2肝细胞内脂质积聚,增加SREBP-1、FAS、ACCα的表达。
[Abstract]:Objective to investigate whether tumor necrosis factor- 伪 (TNF- 伪) can promote lipid accumulation in hepatocytes and its mechanism. Methods HepG2 hepatocytes were divided into blank control group. TNF- 伪 2ng/mL or 20ng / mL 2ng/mL in TNF- 伪 group, palmitic acid group (0.08mmol/L or 0.2 mmol / L) and combined group TNF- 伪 2ng/mL combined with palmitic acid 0.08 mmol / L TNF- 伪 2ng/mL combined with palmitic acid 0.2 mmol / L TNF- 伪 20ng/mL combined with palmitic acid 0.08 mmol / L TNF- 伪 20ng/mL combined with palmitic acid 0.2 mmol / L ~ (-1) for 24 h. TNF- 伪 20ng/mL and 0.08 mmol / L palmitate were selected to observe the accumulation of lipid in HepG2 cells by oil red O staining, and the expression level of SREBP-1FASAACC 伪 in HepG2 cells was detected by real-time fluorescence quantitative PCR and Western blot. 缁撴灉鈶犲崟绾疶NF-伪缁凾G鍚�閲廩TNF-伪2ng/mL缁，

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