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Perilipin 2在小鼠肝脏CGI-58特异性敲除所致肝脏微泡型脂肪变性中的作用
Role of Perilipin 2 in microvesicular hepatic steatosis induced by CGI-58 specific knockout in mice
- 陆军军医大学学报 2024年46卷第15期 页码：1701-1712
- 作者机构：
1.400038 重庆，陆军军医大学(第三军医大学)第一附属医院心血管内科代谢生物钟与心血管疾病中心
2.400038 重庆，陆军军医大学(第三军医大学)第一附属医院老年心脑血管疾病教育部重点实验室
3.400016 重庆，重庆医科大学基础医学院
4.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系病理生理学教研室
5.400038 重庆，陆军军医大学(第三军医大学)第一附属医院全科医学科
- 作者简介：
[ 包鑫余，E-mail：bxytmmu@126.com BAO Xinyu, E-mail: bxytmmu@126.com
[ 张志辉，E-mail：xyzpj@126.com ZHANG Zhihui, E-mail: xyzpj@126.com
- 基金信息：国家自然科学基金面上项目(81873663);重庆市研究生科研创新项目(CYS21515)
DOI：10.16016/j.2097-0927.202405103
中图分类号： R394.3;R575.5;R589.2收稿：2024-05-28，
修回：2024-06-29，
纸质出版：2024-08-15
- 稿件说明：
移动端阅览
张艺馨, 李杰, 万小勤, 等. Perilipin 2在小鼠肝脏CGI-58特异性敲除所致肝脏微泡型脂肪变性中的作用[J]. 陆军军医大学学报, 2024,46(15):1701-1712.
Yixin ZHANG, Jie LI, Xiaoqin WAN, et al. Role of Perilipin 2 in microvesicular hepatic steatosis induced by CGI-58 specific knockout in mice[J]. Journal of Army Medical University, 2024, 46(15): 1701-1712.
张艺馨, 李杰, 万小勤, 等. Perilipin 2在小鼠肝脏CGI-58特异性敲除所致肝脏微泡型脂肪变性中的作用[J]. 陆军军医大学学报, 2024,46(15):1701-1712. DOI： 10.16016/j.2097-0927.202405103.
Yixin ZHANG, Jie LI, Xiaoqin WAN, et al. Role of Perilipin 2 in microvesicular hepatic steatosis induced by CGI-58 specific knockout in mice[J]. Journal of Army Medical University, 2024, 46(15): 1701-1712. DOI： 10.16016/j.2097-0927.202405103.
摘要
目的
2
探究Perilipin 2(
Plin
2)在肝脏CGI-58特异性敲除所致脂肪肝的作用，并比较
Plin
2和
Plin
3在脂滴生成和脂质蓄积的作用效能。
方法
2
将20只7周龄CGI-58
Flox/Flox
小鼠(雄鼠10只，雌鼠10只)根据性别采用完全随机分组法分为对照组(NC组，
n
=5，注射靶向肝脏实现过表达Cre蛋白及对照Micro-RNA的腺相关病毒)和实验组(Mi-KD组，
n
=5，注射靶向肝脏实现过表达Cre蛋白及靶向
Plin
2的Micro-RNA的腺相关病毒)，记录小鼠体质量变化，检测小鼠代谢相关表型和肝脏病理学的差异，RT-qPCR检测小鼠肝脏脂质代谢和胆固醇代谢相关基因的改变；以AML-12小鼠肝细胞作为细胞模型，构建SiRNA实现AML-12细胞
Plin
2/
Plin
3敲低，Bodipy染色和酶比色法检测对比正常培养和OA诱导下
Plin
2/
Plin
3敲低后AML-12细胞脂滴生成和脂质蓄积。
结果
2
Mi-KD小鼠肝脏中的PLIN2蛋白水平降低了99%以上；Mi-KD缓解了CGI-58特异性敲除雌鼠的肝肿大(
P
=0.019 5)和肝细胞损伤(
P
=0.000 4)，降低了组织学NAS评分(
P
=0.000 2)和肝脏甘油三酯含量(
P
=0.016 6)，显著改善了肝脏CGI-58特异性敲除所致的肝脏微泡型脂肪变性；敲低
Plin
2使AML-12细胞内甘油三酯含量呈下降趋势，敲低
Plin
3显著降低AML-12细胞内甘油三酯含量(
P
=0.001 4)；在OA诱导下，
Plin
2/
Plin
3敲低均显著降低OA诱导的AML-12细胞内甘油三酯的蓄积(
P
<
0.05)。
结论
2
肝脏
Plin
2在CGI-58敲除所致的微泡型脂肪变性的发展过程中至关重要，
Plin
2和
Plin
3均参与肝细胞脂滴生成与脂质蓄积，且
Plin
3的作用强于
Plin
2。
Abstract
Objective
2
To explore whether hepatocyte Perilipin-2 (
Plin
2) is involved in the development of fatty liver related to comparative gene identification-58 (CGI-58) deficiency mice and compare the effects of
Plin
2 and
Plin
3 on lipid droplet formation and lipid accumulation.
Methods
2
Based on CGI-58
Flox/Flox
mice as animal model
the adeno-associated viruses targeting mouse liver
CGI-58 knockout and
Plin
2 knockdown were achieved by co-expression Cre protein and micro-RNA targeting
Plin
2 (Mi-KD). Then CGI-58 deficiency mice were used as control (NC) to detect the differences in metabolic phenotype and liver pathology. AML-12 mouse hepatocytes were used as cellular model and interfered with siRNA to achieve
Plin
2/
Plin
3 knockdown in AML-12 cells. Lipid droplet formation and lipid accumulation were compared with Bodipy staining and enzyme colorimetry in basal condition or lipid-overloaded condition (OA inducement) after
Plin
2/
Plin
3 knockdown.
Results
2
Plin
2 knockdown (Mi-KD) reduced PLIN2 protein level by
>
99% in mouse livers. Mi-KD decreased hepatomegaly (
P
=0.019 5) and liver injury (
P
=0.000 4)
while reduced the histological NAS score (
P
=0.000 2) and hepatic triglyceride content (
P
=0.016 6) in the CGI-58 deficiency female mice. Mi-KD prevented microvesicular hepatic steatosis in the CGI-58 deficient female mice.
Plin
3 knockdown significantly reduced the triglyceride content in basal condition of hepa
tocytes (
P
=0.001 4)
and
Plin
2 knockdown just showed a decreased trend.
Plin
2 or
Plin
3 knockdown significantly reduced the triglyceride content separately in lipid-overloaded hepatocytes (
P
<
0.05).
Conclusion
2
Hepatocyte
Plin
2 is essential in the development of microvesicular hepatic steatosis caused by CGI-58 deficiency. Both
Plin
2 and
Plin
3 are involved in lipid droplet formation and lipid accumulation in hepatocytes
and
Plin
3 shows a stronger effect.
关键词
Keywords
references
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