<italic style="font-style: italic">Burkholderia pseudomallei</italic> regulates lipid metabolism in HepG2 cells by inducing autophagy suppression

Mengling TANG; Zhiqiang HU; Siqi YUAN; Chenglong RAO; Yu ZHANG; Jianping XIE; Qian LI; Xuhu MAO

doi:10.16016/j.1000-5404.201906061

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Burkholderia pseudomallei regulates lipid metabolism in HepG2 cells by inducing autophagy suppression

Basic Medicine | 更新时间：2026-01-09

- Burkholderia pseudomallei regulates lipid metabolism in HepG2 cells by inducing autophagy suppression
- Journal of Third Military Medical University Vol. 41, Issue 18, Pages: 1722-1729(2019)
- 作者机构：
  
  1.400715 重庆，西南大学生命科学学院，三峡地区生态环境与生物资源国家重点实验室培育基地，现代生物制药研究所
  2.400038 重庆，陆军军医大学(第三军医大学)药学与检验医学系临床微生物及免疫学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81471914);the General Program of National Natural Science Foundation of China(81772141)
- DOI：10.16016/j.1000-5404.201906061
  CLC： R322.47;R329.28;R378.99
- Received：11 June 2019，
  
  Revised：2019-08-01，
  
  Published：30 September 2019
- 稿件说明：
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Mengling TANG, Zhiqiang HU, Siqi YUAN, et al. Burkholderia pseudomallei regulates lipid metabolism in HepG2 cells by inducing autophagy suppression[J]. Journal of Third Military Medical University, 2019, 41(18): 1722-1729.
DOI：

Mengling TANG, Zhiqiang HU, Siqi YUAN, et al. Burkholderia pseudomallei regulates lipid metabolism in HepG2 cells by inducing autophagy suppression[J]. Journal of Third Military Medical University, 2019, 41(18): 1722-1729. DOI： 10.16016/j.1000-5404.201906061.

摘要

目的

探讨类鼻疽杆菌感染HepG2细胞后诱导的自噬抑制对胞内脂质代谢的调控作用。

方法

类鼻疽杆菌感染HepG2细胞6、12、24 h为感染组，以未感染组作为对照。激光共聚焦显微镜观测各组细胞内脂滴和GFP-自噬点的数量；定量试剂盒检测各组细胞内甘油三酯及胆固醇水平；Western blot检测各组细胞中自噬相关蛋白LC3Ⅱ、p62表达水平。利用自噬抑制剂3-MA、Baf-A1以及自噬激活剂Rapa处理细胞，检测改变宿主细胞自噬水平后，对类鼻疽杆菌感染HepG2细胞后胞内脂质代谢的影响；外源加入游离脂肪酸三醋酸甘油酯，通过细菌胞内计数实验检测类鼻疽杆菌胞内存活情况。

结果

类鼻疽杆菌感染HepG2细胞6、12、24 h后，与未感染组比较，胞内脂滴数量增多，脂质成分甘油三酯和胆固醇水平也随感染时间延长逐渐升高，且在感染24 h后达到最高水平(

0.01)；自噬检测发现，与未感染组比较，LC3Ⅱ蛋白表达随感染时间逐渐下降，p62蛋白表达水平随感染时间逐渐蓄积，胞内自噬小体的数量也随感染时间逐渐减少，且感染24 h后数量最低(

0.01)，提示类鼻疽杆菌感染HepG2细胞后自噬受抑；使用自噬抑制剂3-MA、Baf-A1以及自噬激活剂Rapa处理细胞后，与对照组比较，抑制自噬导致胞内脂质蓄积加重(

0.01)，而激活自噬可以减轻胞内脂质的蓄积(

0.01)；此外，外源补充三醋酸甘油酯促进宿主细胞脂质代谢速率能明显降低类鼻疽杆菌胞内存活量(

0.01)。

结论

类鼻疽杆菌感染HepG2细胞后，通过诱导自噬抑制调控HepG2细胞胞内脂质代谢。

Abstract

Objective

To investigate the regulatory effect of

Burkholderia pseudomallei

(

B. pseudomallei

)-induced autophagy suppression on lipid metabolism in HepG2 cells.

Methods

HepG2 cells infected by

B. pseudomallei

were ob

served for changes in intracellular lipid droplets and GFP-autophagy spots using confocal laser microscopy at 6

24 h following the infection. The levels of triglyceride and cholesterol in the cells were determined using a quantitative detection kit

and the expression levels of autophagy-related proteins LC3Ⅱ and p62 were detected with Western blotting. The effects of pretreatment with the autophagy inhibitors 3-MA and Baf-A1 or the autophagy activator Rapa prior to

B. pseudomallei

infection were examined on lipid metabolism in HepG2 cells. The changes in the intracellular survival of

B. pseudomallei

in response to exogenous free fatty acid triacsin C were examined by counting the viable bacteria in the cells.

Results

B. pseudomallei

infection resulted in significantly increased number of intracellular lipid droplets at 6

12 and 24 h after the infection and progressively increased levels of triglyceride and cholesterol in HepG2 cells over time

which reached the highest level at 24 h after the infection (

0.01). The cells infected by

B. pseudomallei

showed progressively lowered expression of LC3Ⅱ protein and increased expression of p62 protein over time with gradually decreased number of autophagosomes

which reached the lowest level at 24 h after the infection (

0.01)

suggesting suppressed autophagy in HepG2 cells after

B. pseudomallei

infection. Inhibition of autophagy by pretreatment of the cells with 3-MA and Baf-A1 caused significantly increased intracellular lipid accumulation (

0.01)

whereas activation of autophagy by Rapa obviously reduced the accumulation of intracellular lipids (

0.01). The addition of exogenous triacsin C significantly reduced the intracellular survival of

B. pseudomallei

(

0.01).

Conclusion

B. pseudomallei

infection regulates intracellular lipid metabolism in HepG2 cells by inducting autophagy inhibition.

关键词

Keywords

references

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⁰

Burkholderia pseudomallei regulates lipid metabolism in HepG2 cells by inducing autophagy suppression

DOI：10.16016/j.1000-5404.201906061

摘要

Abstract

关键词

Keywords

references