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CD36对炎症因子TNF-α和IL-6诱发HepG2细胞胞内脂质聚集的影响
CD36 knockdown alleviates lipid accumulation induced by tumor necrosis factor-α and interleukin-6 in HepG2 cells
- 第三军医大学学报 2018年40卷第20期 页码：1833-1838
- 作者机构：
400016 重庆，重庆医科大学脂糖代谢性疾病重庆市重点实验室，脂质研究中心
- 作者简介：
[杨萍，E-mail：yang_apple@yeah.net YANG Ping, E-mail:yang_apple@yeah.net
- 基金信息：国家自然科学基金青年科学基金(81400786)
DOI：10.16016/j.1000-5404.201805010
中图分类号： R322.47;R392.3;R589.2收稿：2018-05-02，
修回：2015-06-02，
纸质出版：2018-10-30
- 稿件说明：
移动端阅览
韦莉, 吴俊, 赵蕾, 等. CD36对炎症因子TNF-α和IL-6诱发HepG2细胞胞内脂质聚集的影响[J]. 第三军医大学学报, 2018,40(20):1833-1838.
Li WEI, Jun WU, Lei ZHAO, et al. CD36 knockdown alleviates lipid accumulation induced by tumor necrosis factor-α and interleukin-6 in HepG2 cells[J]. Journal of Third Military Medical University, 2018, 40(20): 1833-1838.
韦莉, 吴俊, 赵蕾, 等. CD36对炎症因子TNF-α和IL-6诱发HepG2细胞胞内脂质聚集的影响[J]. 第三军医大学学报, 2018,40(20):1833-1838. DOI： 10.16016/j.1000-5404.201805010.
Li WEI, Jun WU, Lei ZHAO, et al. CD36 knockdown alleviates lipid accumulation induced by tumor necrosis factor-α and interleukin-6 in HepG2 cells[J]. Journal of Third Military Medical University, 2018, 40(20): 1833-1838. DOI： 10.16016/j.1000-5404.201805010.
摘要
目的
2
探讨炎症因子是否通过上调脂肪酸转运导致HepG2细胞脂质积聚及脂毒性。
方法
2
分别给予HepG2细胞肿瘤坏死因子(tumor necrosis factor-α
TNF-α组: 0.04 mmol/L软脂酸+25 ng/mL TNF-α)和白介素-6(interleukin-6，IL-6组: 0.04 mmol/L软脂酸+20 ng/mL IL-6)刺激处理24 h，酶法检测HepG2细胞内的甘油三酯(triglycercide，TG)水平，ELISA检测细胞内游离脂肪酸(free fatty acid，FFA)含量。利用荧光标记的软脂酸，动态监测细胞对外源性脂肪酸的摄取速率。然后利用小RNA干扰技术，构建低表达脂肪酸转运酶(cluster of differentiation
CD36)的HepG2细胞(CD36 KD HepG2)和对照细胞(NC HepG2)模型，检测CD36低表达时细胞对脂肪酸的摄取速率变化，并检测细胞内活性氧(reactive oxygen species
ROS)及过氧化氢(hydrogen peroxide
H
2
O
2
)的含量。
结果
2
炎症因子能够刺激HepG2细胞TG及FFA的累积，其中TNF-α处理组及IL-6处理组的FFA含量及TG含量都明显增加(
P
<
0.05)，并且能够引起HepG2细胞对脂肪酸的摄取增加(
P
<
0.05)。抑制HepG2细胞中CD36表达，可以减轻胞内脂质积聚(
P
<
0.05)，削弱炎症状态下HepG2细胞对脂肪酸的摄取(
P
<
0.05)，并且能够显著降低细胞内ROS及H
2
O
2
含量(
P
<
0.05)。
结论
2
在肝脏细胞中，抑制CD36的表达能够减少炎症因子TNF-α和IL-6引起的脂肪酸摄取增加，减轻脂质积聚，改善细胞脂毒性；提示CD36可作为脂肪肝等代谢性疾病的潜在治疗靶点。
Abstract
Objective
2
To investigate whether inflammatory cytokines participate in lipid accumulation and lipid toxicity by up-regulating fatty acid transport in HepG2 cells.
Methods
2
HepG2 cells were treated with 25 ng/mL tumor necrosis factor-α (TNF-α) or 20 ng/mL interleukin-6 (IL-6) in the presence of 0.04 mmol/L palmitate for 24 h
followed by enzymatic detection of triglycerides (TG) levels and ELISA to detect intracellular free fatty acid contents. The uptake rate of exogenous fatty acids was monitored using fluorescence-labeled palmitic acid. Small RNA interference technique was used to inhibit the expression of CD36 in HepG2 cells
and the changes in the uptake rate of fatty acids
intracellular reactive oxygen species (ROS)
and hydrogen peroxide (H
2
O
2
) were detected in the cells in response to TNF-α or IL-6 stimulation.
Results
2
The inflammatory cytokines TNF-α and IL-6 stimulated the accumulation of TG and FFA and aggravated intracellular lipid accumulation in HepG2 cells. Treatments of the cells with TNF-α and IL-6 both significantly increased the intracellular FFA and TG contents (
P
<
0.05) and the uptake rate of fatty acids (
P
<
0.05). CD36 knockdown significantly reduced intracellular lipid accumulation (
P
<
0.05)
intake rate of fatty acids (
P
<
0.05)
and intracellular ROS and H
2
O
2
levels in HepG2 cells with TNF-α or IL-6 stimulation (
P
<
0.05).
Conclusions
2
CD36 knockdown can reduce the uptake of fatty acids and lipid accumulation and alleviate lipotoxicity induced by inflammatory cytokines in HepG2 cells
suggesting the potential of CD36 as a therapeutic target for metabolic diseases such as fatty liver.
关键词
Keywords
references
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