摘要
目的:探讨清脂护肝方含药血清对HepG2细胞脂质沉积的影响及可能的作用机制。方法:0.5 mmol/L油酸︰棕榈酸(2︰1)诱导HepG2细胞建立脂肪变性细胞模型,分为空白对照组、模型组、中药血清组(3个浓度:10%、20%、40%)、对照血清组(3个浓度:10%、20%、40%),分别进行细胞培养,油红O染色观察不同组别、不同浓度血清对HepG2细胞脂质沉积的影响,筛选出作用明显的血清浓度再次进行培养,测定细胞的三酰甘油水平,反转录-聚合酶链反应(reverse transcription-polymerase chain reaction, RT-PCR)试验检测脂肪酸合成酶(fatty acid syntherase, FASN)、脂肪酸氧化关键酶肉毒碱棕榈酰基转移酶1A(carnitine paimitoyltransferase 1A, CPT1A)、羟甲基戊二酸单酰辅酶A还原酶(hydroxymethylglutaryl CoA reductase, HMGCR)、固醇调节元件结合蛋白(sterol regulatory element binding transcription factor, SREBF)、过氧化物酶增殖物激活受体α(peroxisome proliferator activated receptor alpha, PPARα)的mRNA水平。结果:(1)0.5 mmol/L油酸︰棕榈酸(2︰1)可成功诱导建立HepG2细胞脂肪变性的模型;(2)油红O染色观察提示20%、40%浓度的清脂护肝方血清可改善脂肪变性HepG2细胞的脂质沉积,40%浓度的效果尤为显著;(3)40%中药血清组三酰甘油水平[(0.077±0.006) mmol/gprot]明显低于模型组[(0.172±0.016) mmol/gprot](P<0.01);(4)模型组细胞的FASN、HMGCR、SREBF的mRNA水平较空白对照组升高,CPT1A、PPARα的mRNA水平较空白对照组降低(P<0.05);40%对照血清组各指标的mRNA水平与模型组比较差异无统计学意义(P>0.05);40%中药血清组FASN、HMGCR、SREBF的mRNA水平明显低于模型组(P<0.05),CPT1A、PPARα的mRNA水平与模型组差异无统计学意义(P>0.05)。结论:清脂护肝方药物血清可减轻脂肪变性的HepG2细胞的脂质沉积,可能是通过下调FASN、HMGCR、SREBF的mRNA水平而实现。
Objective : To research the mechanism of improving HepG2 cell steatosis by Qingzhi Hugan prescription. Methods : Fatty degeneration of HepG2 cell was induced by 0.5 mmol/L oleinic acid and palmitic acid(2︰1), and the experiment was divided into the following groups: the blank control group, the model group, the Qingzhi Hugan prescription serum group(further divided into 3 concentrations: 10%, 20% and 40%) and the control serum group(further divided into 3 concentrations: 10%, 20% and 40%). Cultured cells according to different conditions of different groups, and then used oil red O staining to observe which concentration of the prescription serum significantly improved lipid deposition of HepG2 cell. Then selected the optimal concentration of the prescription serum to culture the HepG2 cells of steatosis, afterwards, examined the triglyceride levels of the cells and utilized reverse transcription-polymerase chain reaction(RT-PCR) method to measure the mRNA levels of fatty acid syntherase(FASN), carnitine paimitoyltransferase 1 A(CPT1 A), hydroxymethylglutaryl CoA reductase(HMGCR), sterol regulatory element binding transcription factor(SREBF) and peroxisome proliferator activated receptor alpha(PPARα). Results :(1)Used oleinic acid and palmitic acid(2︰1) could successfully induce fat degeneration of HepG2 cell.(2)Oil red O staining suggested that the prescription serum of 20% and 40% could improve lipid deposition of fatty degeneration HepG2 cells, and the effect of 40% was particularly significant.(3)Compared with the model group[(0.172±0.016) mmol/gprot], the level of triglyceride [(0.077±0.006) mmol/gprot] was significantly decreased in 40% of the prescription serum group(P<0.01).(4)The mRNA levels of FASN, HMGCR and SREBF in the model group were higher than those in the blank control group, inversely, the mRNA levels of CPT1 A and PPARα were low(P<0.05). There was no significant difference of all the mRNA levels between the model group and the 40% control serum group(P>0.05). The mRNA levels of FASN, HMGCR and SREBF in the 40% prescription serum group were significantly lower than those in the model group(P<0.05), and the mRNA levels of CPT1 A and PPARα were not significantly different from those in the model group(P>0.05). Conclusions : Qingzhi Hugan prescription serum could reduce lipid deposition of fatty degeneration HepG2 cell, and it might be achieved by lowering the mRNA levels of FASN, HMGCR and SREBF.
作者
罗蕾蕾
邵建国
卞兆连
汪洋
LUO Leilei;SHAO Jianguo;BIAN Zhaolian;WANG Yang(Department of Gastroenterology,the Third People's Hospital of Nantong,Jiangsu Province,Nantong 226006;Clinical Medicine of Medical College of Nantong University)
出处
《南通大学学报（医学版）》
2019年第4期241-245,共5页
Journal of Nantong University(Medical sciences)
基金
南通市青年医学人才科研基金资助项目(WQ2016008)
