TNF-α促進(jìn)HepG2肝細(xì)胞脂質(zhì)積聚及其機(jī)制的初步研究
發(fā)布時(shí)間:2018-04-27 08:07
本文選題:TNF-α + 軟脂酸 ; 參考:《第三軍醫(yī)大學(xué)學(xué)報(bào)》2013年11期
【摘要】:目的探討腫瘤壞死因子-α(tumor necrosis factor-α,TNF-α)是否能夠促進(jìn)肝細(xì)胞脂質(zhì)積聚,并對(duì)其機(jī)制進(jìn)行初步探討。方法將HepG2肝細(xì)胞分為空白對(duì)照組、單純TNF-α組(TNF-α2ng/mL或20ng/mL)、軟脂酸組(軟脂酸0.08mmol/L或0.2mmol/L)及聯(lián)合組(TNF-α2ng/mL聯(lián)合軟脂酸0.08mmol/L、TNF-α2ng/mL聯(lián)合軟脂酸0.2mmol/L、TNF-α20ng/mL聯(lián)合軟脂酸0.08mmol/L、TNF-α20ng/mL聯(lián)合軟脂酸0.2mmol/L),處理24h,應(yīng)用化學(xué)酶促-比色法定量檢測(cè)細(xì)胞內(nèi)TG含量。進(jìn)一步選取TNF-α20ng/mL和軟脂酸0.08mmol/L,通過(guò)油紅O染色觀察HepG2細(xì)胞內(nèi)脂質(zhì)積聚情況;實(shí)時(shí)熒光定量PCR和Western blot檢測(cè)HepG2細(xì)胞SREBP-1、FAS、ACCα的表達(dá)水平。結(jié)果①單純TNF-α組TG含量[TNF-α2ng/mL組(0.344±0.093)μg/μg、TNF-α20ng/mL組(0.329±0.068)μg/μg]分別較空白對(duì)照組[(0.192±0.048)μg/μg]顯著升高(P0.05);聯(lián)合組[TNF-α2ng/mL聯(lián)合軟脂酸0.08mmol/L組(0.451±0.096)μg/μg、TNF-α2ng/mL聯(lián)合軟脂酸0.2mmol/L組(0.821±0.257)μg/μg、TNF-α20ng/mL聯(lián)合軟脂酸0.08mmol/L組(1.032±0.286)μg/μg、TNF-α20ng/mL聯(lián)合軟脂酸0.2mmol/L組(2.134±1.049)μg/μg]分別較軟脂酸組[軟脂酸0.08mmol/L組(0.247±0.069)μg/μg、軟脂酸0.2mmol/L組(0.341±0.031)μg/μg]顯著升高(P0.05);②油紅O染色進(jìn)一步顯示,TNF-α促進(jìn)肝細(xì)胞內(nèi)脂質(zhì)積聚。③實(shí)時(shí)熒光定量PCR和Western blot檢測(cè)結(jié)果顯示單純TNF-α組與空白對(duì)照組相比,HepG2細(xì)胞SREBP-1、FAS、ACCα的表達(dá)均增加(P0.05);聯(lián)合組與軟脂酸組相比,肝細(xì)胞內(nèi)SREBP-1、FAS、ACCα的表達(dá)水平明顯上調(diào)(P0.05)。結(jié)論TNF-α促進(jìn)HepG2肝細(xì)胞內(nèi)脂質(zhì)積聚,增加SREBP-1、FAS、ACCα的表達(dá)。
[Abstract]:Objective to investigate whether tumor necrosis factor- 偽 (TNF- 偽) can promote lipid accumulation in hepatocytes and its mechanism. Methods HepG2 hepatocytes were divided into blank control group. TNF- 偽 2ng/mL or 20ng / mL 2ng/mL in TNF- 偽 group, palmitic acid group (0.08mmol/L or 0.2 mmol / L) and combined group TNF- 偽 2ng/mL combined with palmitic acid 0.08 mmol / L TNF- 偽 2ng/mL combined with palmitic acid 0.2 mmol / L TNF- 偽 20ng/mL combined with palmitic acid 0.08 mmol / L TNF- 偽 20ng/mL combined with palmitic acid 0.2 mmol / L ~ (-1) for 24 h. TNF- 偽 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 偽 in HepG2 cells was detected by real-time fluorescence quantitative PCR and Western blot. 緇撴灉鈶犲崟綰疶NF-偽緇凾G鍚噺[TNF-偽2ng/mL緇,
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