脂肪细胞Npy4r促进高脂饮食诱导肥胖

doi:10.16288/j.yczz.22-302

遗传 ›› 2023, Vol. 45 ›› Issue (2): 144-155.doi: 10.16288/j.yczz.22-302

脂肪细胞Npy4r促进高脂饮食诱导肥胖

王澜(), 曾帆, 黄荣凤, 林树, 张志辉, 李旻典()

陆军军医大学第一附属医院心血管内科，代谢生物钟与心血管病中心，重庆 400038

收稿日期:2022-10-13 修回日期:2023-01-30 出版日期:2023-02-20 发布日期:2023-02-03
通讯作者: 李旻典 E-mail:770944509@qq.com;mindianli@tmmu.edu.cn
作者简介:王澜，硕士，研究实习员，研究方向：代谢生物钟与心血管疾病。E-mail: 770944509@qq.com
基金资助:
国家自然科学基金青年科学基金项目(81900776)

Adipocyte reconstitution of Npy4r gene in Npy4r silenced mice promotes diet-induced obesity

Lan Wang(), Fan Zeng, Rongfeng Huang, Shu Lin, Zhihui Zhang, Min-Dian Li()

Department of Cardiovascular Medicine, Center for Circadian Metabolism and Cardiovascular Disease, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, 400038, China

Received:2022-10-13 Revised:2023-01-30 Online:2023-02-20 Published:2023-02-03
Contact: Li Min-Dian E-mail:770944509@qq.com;mindianli@tmmu.edu.cn
Supported by:
the National Natural Science Foundation of China(81900776)

摘要/Abstract

摘要：

脂肪组织的神经支配与调节在能量代谢稳态的维持中发挥重要作用。神经肽Y (neuropeptide Y, NPY)及其脂肪细胞受体信号通路促进高脂饮食诱导的肥胖，其中NPY受体1 (NPY receptor Y1, NPY1R)与受体2(NPY2R)是主要的NPY外周受体。NPY受体4 (NPY4R)也在脂肪组织表达，然而尚不清楚其是否参与肥胖的发生发展机制。本研究建立了NPY及其受体的免疫荧光成像技术和脂肪细胞回复性表达Npy4r小鼠。根据对不同部位脂肪组织的荧光显微术观察，发现NPY在肩胛间棕色脂肪和皮下脂肪的围绕血管区域以点状形式表达，NPY系统的各受体在脂肪组织的空间分布上具有明显的组织特异性：NPY1R在棕色脂肪、主动脉周围脂肪和性腺脂肪较为富集，NPY2R在棕色脂肪和主动脉周围脂肪较为富集，NPY4R在棕色脂肪与性腺脂肪较为富集。继而通过比较脂肪细胞回复性表达Npy4r小鼠与全身Npy4r基因静默小鼠在高脂喂食下的体重与糖代谢，发现脂肪细胞Npy4r促进高脂饮食诱导的肥胖(P < 0.0001)。本研究明确了NPY及其受体NPY1R、NPY2R和NPY4R在不同部位脂肪组织的蛋白水平与分布，进而用脂肪细胞特异性地回复表达小鼠模型揭示脂肪细胞Npy4r具有促进高脂饮食诱导肥胖的作用。

关键词: 条件性基因回复表达, loxP-STOP-loxP, 脂肪细胞, 脂肪组织, 肥胖

Abstract:

Neural regulation of adipose tissue is crucial in the homeostasis of energy metabolism. Adipose tissue neuropeptide Y (NPY) and its receptors contribute to the development of diet-induced obesity. NPY1R and NPY2R are major receptors for NPY in peripheral tissues including the adipose tissue. NPY receptor 4 (Npy4r) gene is expressed in adipose tissue. However, it is unknown whether Npy4r is involved in the development of diet-induced obesity. Here, we established an immunofluorescence microscopy technique and generated an adipocyte-reconstituted Npy4r gene knockout mouse. Among six adipose depots, we found that NPY is highly expressed around the vasculature in a dot-like fashion in interscapular brown fat and subcutaneous fat, and NPY receptors are expressed in a depot-specific manner. NPY1R is highly expressed in epidydimal fat, interscapular and peri-aortic brown fat, NPY2R in both interscapular and peri-aortic brown fat, and NPY4R in both brown fat and epidydimal fat. Next, we showed that adipocyte-reconstituted expression of Npy4r promoted diet-induced obesity in mice (P < 0.0001). Overall, this study defines the abundance and distribution of NPY and its receptors 1, 2, and 4 in mouse adipose depots, and demonstrates in an adipocyte-reconstituted gene knockout model that adipocyte Npy4r is sufficient to promote diet-induced obesity.

Key words: Conditionally reconstituted gene knockout, loxP-STOP-loxP, adipocyte, adipose tissue, obesity

王澜, 曾帆, 黄荣凤, 林树, 张志辉, 李旻典. 脂肪细胞Npy4r促进高脂饮食诱导肥胖[J]. 遗传, 2023, 45(2): 144-155.

Lan Wang, Fan Zeng, Rongfeng Huang, Shu Lin, Zhihui Zhang, Min-Dian Li. Adipocyte reconstitution of Npy4r gene in Npy4r silenced mice promotes diet-induced obesity[J]. Hereditas(Beijing), 2023, 45(2): 144-155.

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引物序列	WT	Adp-Y4	Y4-KO
F1: 5′-CAGGCTGCATCTCTGAAGTAGGC-3′	131 bp	165 bp
R1: 5′-TGGATTGGTCCCATTCTTACCCT-3′	131 bp	165 bp
F2: 5′-CCATGAGATATAAGCAGCCAAGAAC-3′			258 bp
R2: 5′-GCTAAAGCGCATGCTCCAGA-3′			258 bp

脂肪细胞Npy4r促进高脂饮食诱导肥胖

Adipocyte reconstitution of Npy4r gene in Npy4r silenced mice promotes diet-induced obesity

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[2]	曾帆, 王澜, 万小勤, 黄荣凤, 张志辉, 李旻典. 瘦素基因启动的新型脂肪细胞表达Cre工具小鼠的构建[J]. 遗传, 2022, 44(10): 950-957.
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