miR-451通过AMPK信号通路调控3T3-L1前脂肪细胞分化的机制研究

doi:10.11958/20253387

天津医药 ›› 2026, Vol. 54 ›› Issue (8): 785-789.doi: 10.11958/20253387

• 细胞与分子生物学 • 下一篇

miR-451通过AMPK信号通路调控3T3-L1前脂肪细胞分化的机制研究

曹参(), 谢琨, 曾维利^△()

郴州市第一人民医院中心医院综合病室（邮编423000）

收稿日期:2025-11-17 修回日期:2026-04-13 出版日期:2026-08-15 发布日期:2026-08-07
通讯作者: △E-mail：13607359506@163.com
作者简介:曹参（1990），女，主治医师，主要从事糖尿病诊治方面研究。E-mail：zgzz_2025@163.com
基金资助:
湖南省卫生健康委科研计划项目(D202303069026)

The mechanism of miR-451 on regulating differentiation of 3T3-L1 preadipocytes through AMPK signaling pathway

CAO Can(), XIE Kun, ZENG Weili^△()

Comprehensive Ward, Central Hospital of Chenzhou First People's Hospital, Chenzhou 423000, China

Received:2025-11-17 Revised:2026-04-13 Published:2026-08-15 Online:2026-08-07
Contact: △E-mail: 13607359506@163.com

曹参, 谢琨, 曾维利. miR-451通过AMPK信号通路调控3T3-L1前脂肪细胞分化的机制研究[J]. 天津医药, 2026, 54(8): 785-789.

CAO Can, XIE Kun, ZENG Weili. The mechanism of miR-451 on regulating differentiation of 3T3-L1 preadipocytes through AMPK signaling pathway[J]. Tianjin Medical Journal, 2026, 54(8): 785-789.

摘要/Abstract

摘要：

目的探讨miR-451通过腺苷酸活化蛋白激酶（AMPK）信号通路调控3T3-L1前脂肪细胞分化的作用。方法将3T3-L1细胞随机分为Control组、阴性对照（NC）组、miR-451 mimics组和miR-451 inhibitor组。采用实时荧光定量聚合酶链式反应（qRT-PCR）检测miR-451表达，油红O染色观察脂滴形成情况，萤光素酶报告基因试验验证miR-451与钙结合蛋白39（CAB39）的靶向关系，Western blot检测AMPK、哺乳动物雷帕霉素靶蛋白（mTOR）表达。构建AMPK表达下调的3T3-L1细胞，分为NC-N.A组、miR-451 mimics-N.A组和miR-451 inhibitor-N.A组，检测甘油三酯（TG）水平。结果 3T3-L1细胞诱导分化为成熟脂肪细胞7 d后，miR-451表达升高（P<0.05）。与Control组和NC组相比，miR-451 mimics组miR-451表达水平升高，油红O染色显示脂滴体积增大，染色加深；miR-451 inhibitor组则呈现相反变化（P<0.05）。双萤光素酶报告实验证实CAB39是miR-451的靶基因。Western blot检测结果显示，与Control组、NC组相比，miR-451 mimics组p-AMPK表达下降，p-mTOR表达升高（P<0.05）；miR-451 inhibitor组p-AMPK表达升高，p-mTOR表达下降（P<0.05）。在AMPK敲低的3T3-L1细胞中上调或下调miR-451不影响TG的生成。结论 miR-451可促进3T3-L1前脂肪细胞分化，该过程与miR-451抑制AMPK信号通路有关。

关键词: 微RNAs, 脂细胞, miR-451, 腺苷酸激酶

Abstract:

Objective To investigate the role and mechanism of miR-451 in regulating the differentiation of 3T3-L1 preadipocytes through the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway. Methods 3T3-L1 cells were randomly divided into the control group, the NC group, the miR-451 mimics group and the miR-451 inhibitor group. The expression of miR-451 was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Lipid droplet formation was observed by Oil Red O staining. The targeting relationship between miR-451 and calcium-binding protein 39 (CAB39) was verified by dual-luciferase reporter gene assay. The protein expressions of AMPK and mammalian target of rapamycin (mTOR) were detected by Western blot assay. 3T3-L1 cells with AMPK knockdown were constructed and divided into the NC-N.A group, the miR-451 mimics-N.A group and the miR-451 inhibitor-N.A group. Triglyceride (TG) levels were measured. Results After 7 days of induced differentiation of 3T3-L1 cells into mature adipocytes, the expression of miR-451 was significantly increased (P<0.05). Compared with the control group volume and the NC group, the miR-451 mimics group showed significantly increased miR-451 expression, larger lipid droplets volume and deeper Oil Red O staining (P<0.05). In contrast, the miR-451 inhibitor group exhibited the opposite changes (P<0.05). The dual-luciferase reporter assay confirmed that CAB39 was a direct target gene of miR-451. Western blot results showed that, compared with the control group and the NC group, p-AMPK expression decreased, and p-mTOR expression increased in the miR-451 mimics group (P<0.05), while p-AMPK expression increased and p-mTOR expression decreased in the miR-451 inhibitor group (P<0.05). In AMPK-knockdown 3T3-L1 cells, neither overexpression nor inhibition of miR-451 affected TG production. Conclusion miR-451 can promote the differentiation of 3T3-L1 preadipocytes, and this process is associated with the inhibition of the AMPK signaling pathway by miR-451.

Key words: microRNAs, adipocyte, miR-451, adenylate kinase

中图分类号:

R587.1

图/表 7

图1 miR-451与CAB39 mRNA的3'-UTR区碱基互补配对

Fig.1 Base complementary pairing of miR-451 and CAB39 mRNA in 3'-UTR region

图2 miR-451在3T3-L1前脂肪细胞分化过程中的表达情况 a与0 d比较，P<0.05。

Fig.2 Expression of miR-451 in the differentiation process of 3T3-L1 preadipocytes

图3 各组miR-451表达情况 a与Control组比较，b与NC组比较，P<0.05。

Fig.3 Expression of miR-451 in each group

图4 miR-451对3T3-L1前脂肪细胞分化的影响（油红O染色，×200） a与Control组比较，b与NC组比较，P<0.05。

Fig.4 Effects of miR-451 on differentiation of 3T3-L1 preadipocytes (Oil Red O staining, ×200)

图5 萤光素酶报告基因试验检测各组细胞的萤光素酶活性

Fig.5 Detection of cell luciferase activity by luciferase reporter gene assay in each group

图6 各组AMPK信号通路蛋白表达 A：Control组，B：NC组，C：miR-451 mimics组，D：miR-451 inhibitor组；n=3，FAMPK=2.684，P=0.086；Fp-AMPK=139.327，P<0.01；FmTOR=0.922，P=0.410；Fp-mTOR=680.536，P<0.01；a与Control组比较，b与NC组比较，P<0.05。

Fig. 6 Expressions of AMPK signaling pathway proteins in each group

图7 Western blot验证3T3-L1前脂肪细胞AMPK敲低效果 A：shRNA-AMPK组；B：shRNA-NC组。

Fig.7 Western blot verification of AMPK knockdown effect in 3T3-L1 preadipocytes

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miR-451通过AMPK信号通路调控3T3-L1前脂肪细胞分化的机制研究

The mechanism of miR-451 on regulating differentiation of 3T3-L1 preadipocytes through AMPK signaling pathway

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