棕榈酸通过上调ACC/FAS/DGAT2通路表达诱导BRL 3A细胞脂肪变性

doi:10.11843/j.issn.0366-6964.2019.12.018

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (12): 2537-2544.doi: 10.11843/j.issn.0366-6964.2019.12.018

棕榈酸通过上调ACC/FAS/DGAT2通路表达诱导BRL 3A细胞脂肪变性

杨玲^1,2,3, 杨汶菱^1,2,3, 张绘艳^1,2,3, 卞建春^1,2,3, 刘宗平^1,2,3, 袁燕^1,2,3, 邹辉^1,2,3, 顾建红^1,2,3*, 刘学忠^1,2,3*

1. 扬州大学兽医学院, 扬州 225009;
2. 江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009;
3. 扬州大学教育部农业与农产品安全国际合作联合实验室, 扬州 225009

收稿日期:2019-07-17 出版日期:2019-12-23 发布日期:2019-12-20
通讯作者: 顾建红,主要从事动物营养代谢病研究,Tel:0514-87979042,E-mail:jhgu2005@163.com;刘学忠,主要从事动物营养代谢病研究,Tel:0514-87979042,E-mail:liuxuezhong68@163.com
作者简介:杨玲(1995-),女,广东五华人,硕士,主要从事动物营养代谢病研究,E-mail:1825231252@163.com
基金资助:
国家重点研发计划（2016YFD0501206）；江苏省高校优势学科建设工程资助项目（PAPD）

Palmitic Acid Induced Steatosis in BRL 3A Cells by Up-regulating the Expression of ACC/FAS/DGAT2 Pathway

YANG Ling^1,2,3, YANG Wenling^1,2,3, ZHANG Huiyan^1,2,3, BIAN Jianchun^1,2,3, LIU Zongping^1,2,3, YUAN Yan^1,2,3, ZOU Hui^1,2,3, GU Jianhong^1,2,3*, LIU Xuezhong^1,2,3*

1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
2. Jiangsu Co-innovation Center for Prevention and Contrlol of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China;
3. Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, Yangzhou 225009, China

Received:2019-07-17 Online:2019-12-23 Published:2019-12-20

摘要/Abstract

摘要： 为探究棕榈酸诱导BRL 3A细胞脂肪变性的机制，以不同浓度的PA（0、0.2、0.4、0.6 mmol·L^-1）处理BRL 3A细胞24 h，用CCK-8法、RTCA技术检测细胞增殖情况；油红O染色，观察细胞脂滴生成情况；DAPI/F-actin双染，观察细胞核及骨架形态；采用比色法测定TG含量；采用RT-PCR检测脂肪合成相关基因Acaca、Fasn、Dgat2转录水平；采用Western blot检测脂肪合成关键蛋白ACC、FAS、SCD1、GPAM、DGAT2表达量。结果显示：与对照组相比，0.2、0.4 mmol·L^-1 PA对细胞增殖无影响，0.6 mmol·L^-1 PA抑制细胞增殖；油红O染色结果显示0.4 mmol·L^-1 PA处理细胞24 h，脂滴大量蓄积，发生明显脂肪变性；随PA浓度升高，细胞核发生皱缩、变形、碎裂，细胞骨架被破坏，微丝断裂；TG含量极显著增加（P<0.01）；不同浓度PA（0.2、0.4、0.6 mmol·L^-1）处理组脂肪合成关键基因Acaca、Fasn、Dgat2 mRNA转录水平均显著升高（P<0.05），ACC、FAS、SCD1、GPAM、DGAT2蛋白表达水平均显著升高（P<0.05）；0.6 mmol·L^-1 PA组与0.4 mmol·L^-1 PA组相比，SCD1蛋白表达水平显著降低（P<0.05）。高浓度棕榈酸抑制BRL 3A细胞增殖，对细胞核及骨架产生损伤；棕榈酸诱导细胞发生脂滴蓄积，TG增加，通过上调脂肪合成关键基因Acaca、Fasn、Dgat2 mRNA转录水平及ACC/FAS/DGAT2通路蛋白表达水平诱导细胞发生脂肪变性。

关键词: 棕榈酸, 脂肪变性, 乙酰辅酶A羧化酶, 二酰基甘油酰基转移酶2, BRL 3A

Abstract: This paper will focus on the mechanism of palmitic acid-induced BRL 3A cell steatosis. BRL 3A cells were treated with different concentrations of PA (0, 0.2, 0.4, 0.6 mmol·L^-1) for 24 h. The cell proliferation was detected by using CCK-8 method and RTCA technique, the cell lipid droplet formation was observed by using oil red O staining, the nucleus and cytoskeleton morphology were observed by using DAPI/F-actin double staining, the TG content was determined by colorimetry. The transcriptional levels of Acaca, Fasn and Dgat2 were detected by RT-PCR. The expressions of ACC, FAS, SCD1, GPAM and DGAT2 were detected by Western Blot. In result, compared with the control group, the 0.2, 0.4 mmol·L^-1 PA group had no effect on cell proliferation and 0.6 mmol·L^-1 PA inhibited cell proliferation. Oil red O staining showed that a large amount of lipid droplets accumulated in the 0.4 mmol·L^-1 PA group, and obvious fatty degeneration occurred. With the increase of PA concentration, the nucleus shrinks, deforms and fragments; the cytoskeleton was destroyed and the microfilament breaks. With the increase of PA concentration, the TG content increased significantly (P<0.01). The transcriptional levels of Acaca, Fasn and Dgat2 mRNA in the lipid synthesis genes of different concentrations of PA (0.2, 0.4, 0.6 mmol·L^-1) were significantly increased (P<0.05). The protein expression levels of ACC, FAS, SCD1, GPAM and DGAT2 were significantly increased (P<0.05). SCD1 protein expression was significantly lower in the 0.6 mmol·L^-1 PA group compared with the 0.4 mmol·L^-1 PA group (P<0.05). High concentration of palmitic acid inhibited the proliferation of BRL 3A cells and damaged the nucleus and skeleton. Palmitic acid induced the accumulation of lipid droplets and the increase of TG, and induced steatosis by up-regulating the transcription levels of Acaca, Fasn, Dgat2 mRNA and the expression of ACC/FAS/DGAT2 pathway proteins.

Key words: palmitic acid, steatosis, acetyl-CoA carboxylase, diacylglycerol acyltransferase 2, BRL 3A

中图分类号:

S852.33

杨玲, 杨汶菱, 张绘艳, 卞建春, 刘宗平, 袁燕, 邹辉, 顾建红, 刘学忠. 棕榈酸通过上调ACC/FAS/DGAT2通路表达诱导BRL 3A细胞脂肪变性[J]. 畜牧兽医学报, 2019, 50(12): 2537-2544.

YANG Ling, YANG Wenling, ZHANG Huiyan, BIAN Jianchun, LIU Zongping, YUAN Yan, ZOU Hui, GU Jianhong, LIU Xuezhong. Palmitic Acid Induced Steatosis in BRL 3A Cells by Up-regulating the Expression of ACC/FAS/DGAT2 Pathway[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50(12): 2537-2544.

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棕榈酸通过上调ACC/FAS/DGAT2通路表达诱导BRL 3A细胞脂肪变性

Palmitic Acid Induced Steatosis in BRL 3A Cells by Up-regulating the Expression of ACC/FAS/DGAT2 Pathway

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