本文選題：甲醛 + 肝細(xì)胞��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:通過(guò)觀察甲醛(FA)對(duì)HepG2細(xì)胞內(nèi)外甘油三酯(TG)和游離膽固醇(FC)含量以及脂代謝相關(guān)基因表達(dá)水平的影響,來(lái)探討甲醛致肝損傷的可能機(jī)制。方法:以0.004、0.02、0.1 mmol/L FA分別處理人肝癌HepG2細(xì)胞24和48 h,采用POD酶法測(cè)定細(xì)胞內(nèi)外TG和FC含量,采用QRT-PCR法檢測(cè)細(xì)胞內(nèi)與TG代謝相關(guān)基因:固醇調(diào)節(jié)元件結(jié)合蛋白1(SREBP-1)、二脂酰甘油�；D(zhuǎn)移酶2(DGAT2)、過(guò)氧化物酶體增殖物激活受體(PPAPα)、酯酰肉堿轉(zhuǎn)移酶1(CPT1)、微粒體甘油三酯轉(zhuǎn)運(yùn)蛋白(MTTP)、載脂蛋白B100(apoB100)、載脂蛋白E(apoE)、羧酸脂酶3(CES3)、二脂酰甘油酰基轉(zhuǎn)移酶1(DGAT1)、包被蛋白II(COPⅡ)、Sar1b和與FC代謝相關(guān)基因:固醇調(diào)節(jié)元件結(jié)合蛋白2(SREBP-2)、SREBP裂解激活蛋白(SCAP)、胰島素誘導(dǎo)基因1(Insig1)、羥甲基戊二酰輔酶A還原酶(HMGCR)、低密度脂蛋白受體(LDLR)的表達(dá)水平。結(jié)果:第一部分:(1)與陰性對(duì)照組相比,甲醛各劑量組染毒48 h可使HepG2細(xì)胞內(nèi)TG含量明顯減少(P0.05),染毒24 h可使上清中TG含量明顯增加(P0.05);(2)與陰性對(duì)照組相比,染毒48 h,甲醛各劑量組細(xì)胞內(nèi)PPAPα基因表達(dá)水平明顯降低(P0.05);染毒24 h,0.004 mmol/L FA組或染毒48 h,0.1 mmol/L FA組細(xì)胞內(nèi)CPT1基因表達(dá)水平明顯降低(P0.05);(3)與陰性對(duì)照組相比,染毒24 h,0.004 mmol/L FA組或染毒48 h,甲醛各劑量組細(xì)胞內(nèi)MTTP基因表達(dá)水平明顯升高(P0.05);染毒24 h,甲醛各劑量組或染毒48 h,0.004 mmol/L FA組細(xì)胞內(nèi)apoB100基因表達(dá)水平明顯升高(P0.05)。第二部分:(1)與陰性對(duì)照組相比,甲醛各劑量組染毒24和48 h均可使HepG2細(xì)胞內(nèi)FC含量明顯增加(P0.05),且0.004和0.02 mmol/L FA染毒48 h還可使上清中FC含量明顯減少(P0.05);(2)與陰性對(duì)照組相比,染毒24 h,甲醛各劑量組細(xì)胞內(nèi)HMGCR基因表達(dá)水平明顯升高(P0.05),染毒48 h,0.1 mmol/L FA組細(xì)胞內(nèi)HMGCR基因表達(dá)水平明顯降低(P0.05);(3)與陰性對(duì)照組相比,染毒48 h,0.02和0.1 mmol/L FA組細(xì)胞內(nèi)LDLR基因表達(dá)水平明顯升高(P0.05)。結(jié)論:(1)甲醛可能通過(guò)減少HepG2細(xì)胞內(nèi)PPARα及CPT1基因的表達(dá)來(lái)抑制肝臟脂肪酸的β-氧化,同時(shí)可能通過(guò)增加MTTP和apoB100基因的表達(dá)來(lái)促進(jìn)VLDL的分泌外排。(2)短期內(nèi)甲醛可能通過(guò)增加HepG2細(xì)胞內(nèi)HMGCR基因的表達(dá)來(lái)促進(jìn)FC的合成,隨著作用時(shí)間的延長(zhǎng)可能通過(guò)增加LDLR基因的表達(dá)而使細(xì)胞內(nèi)FC水平升高,進(jìn)而反饋抑制HMGCR基因的表達(dá)。本課題為山西省留學(xué)回國(guó)人員科技活動(dòng)擇優(yōu)資助項(xiàng)目“甲醛對(duì)肝臟脂肪代謝的影響及其可能機(jī)制(編號(hào):晉人社廳函[2014]376號(hào))”。
[Abstract]:Aim: to investigate the possible mechanism of hepatic injury induced by formaldehyde by observing the effects of formaldehyde on the content of triglyceride (TG) and free cholesterol (FCC) in HepG2 cells and the expression of genes related to lipid metabolism in HepG2 cells. Methods: human hepatoma HepG2 cells were treated with 0.004 and 0.02mol / L FA for 24 h and 48 h, respectively. The contents of TG and FC in HepG2 cells were measured by POD enzymatic assay. QRT-PCR was used to detect the genes related to TG metabolism in cells: steroid regulatory element binding protein 1 (SREBP-1), diacylglycerol acyltransferase (2) DGAT-2, peroxisome proliferator activator receptor (PPAP 偽), esterylcarnitine transferase (1) CPT _ (1), microsomal triglyceride transposition (TG). Transporter MTTPN, apolipoprotein B100 apo B100, apolipoprotein EapoEN, carboxylidase 3Ces3, diacylglyceryltransferase 1n DGAT1, encapsulated protein II cop 鈪，

本文編號(hào)：2040254