Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells

Wei ZHOU; Li MA; Ying WAN; Fuxiang LI

doi:10.16016/j.1000-5404.201802048

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Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells

更新时间：2026-01-09

- Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells
- Journal of Third Military Medical University Vol. 40, Issue 11, Pages: 954-958(2018)
- 作者机构：
- 作者简介：
- 基金信息：
  
  Supported by the National Nature Science Foundation for Young Scholars of China(31700725)
- DOI：10.16016/j.1000-5404.201802048
  CLC：
- Received：16 February 2018，
  
  Published：2018-06
- 稿件说明：
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Wei ZHOU, Li MA, Ying WAN, et al. Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells[J]. Journal of Third Military Medical University, 2018, 40(11): 954-958.
DOI：

Wei ZHOU, Li MA, Ying WAN, et al. Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells[J]. Journal of Third Military Medical University, 2018, 40(11): 954-958. DOI： 10.16016/j.1000-5404.201802048.

摘要

Abstract

Objective

To determine the effect of regulation of Rab32 on the lipid metabolism in murine hepatic alpha mouse liver 12 (AML12) cells.

Methods

Lentivirus expressing FCD-EYFP-Rab32 was transfected into AML12 cells for overexpression of Rab32

and CRISPR/Cas9 gene knockout technology was used to knockout Rab32 in the cells. Untreated AML12 cells were used as normal control. The protein level of Rab32 was determined by Western blotting. The intracellular lipid droplets were observed by confocal microscopy after Nile red fluorescent staining. The contents of intracellular triglyceride (TG) and total cholesterol (TC) were measured by corresponding quantitation kits. Cell apoptosis was assayed by flow cytometry.

Results

Compared with the normal control cells

the protein level of Rab32 was significantly increased in the overexpressed cells but decreased in the knockout cells (

0.01). The number (21.91±14.32

33.89±17.31

0.01) and size (16.82±14.37

22.27±14.10 μm

0.01) of lipid droplets per cell were significantly decreased in the overexpressed cells when compared with the untreated cells

while the intracellular contents of TG (104.03±12.28

130.94±4.21 nmol

0.01) and TC (46.92±1.26

81.11±0.65 μg

0.01) were decreased. Whereas

the knockout cells had larger lipid droplet number (58.23±42.28

0.01) and total area (53.31±36.33 μm

0.01)

and higher intracellular TG (159.03±8.85 nmol

0.01) and TC (93.38±3.33 μg

0.01). No obvious difference was found in cell apoptosis among the 3 groups of cells (

0.05).

Conclusion

Rab32 promotes the lipid metabolism in the AML12 cells

and the molecule may be regarded as a new target in treatment of nonalcoholic fatty liver disease.

关键词

Keywords

references

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