牛磺熊去氧胆酸通过抑制内质网应激诱导的凋亡促进类鼻疽菌的清除

赵光强; 南栋琪; 袁思琪; 饶承龙; 邢贞泉; 王彬; 方瑶; 毛旭虎; 李倩

doi:10.16016/j.2097-0927.202304046

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牛磺熊去氧胆酸通过抑制内质网应激诱导的凋亡促进类鼻疽菌的清除

基础医学 | 更新时间：2026-01-09

- 牛磺熊去氧胆酸通过抑制内质网应激诱导的凋亡促进类鼻疽菌的清除
- TUDCA promotes intracellular clearance of Burkholderia pseudomallei by inhibiting endoplasmic reticulum stress-induced apoptosis in RAW264.7 cells
- 陆军军医大学学报 2024年46卷第3期页码：225-231
- 作者机构：
  
  1.572022 海南三亚，三亚市人民医院(四川大学华西三亚医院)呼吸内科
  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与免疫学教研室
  3.430070 武汉，解放军中部战区总医院呼吸与危重症医学科
  4.400038 重庆，陆军军医大学(第三军医大学)创伤与化学中毒全国重点实验室
- 作者简介：
  
  [李倩，E-mail：liqianjane@163.com LI Qian, E-mail: liqianjane@163.com
- 基金信息：
  
  国家自然科学基金面上项目(32270190);海南省自然科学基金面上项目(821MS162)
- DOI：10.16016/j.2097-0927.202304046
  中图分类号： R285.5;R329.28;R378.99
- 收稿：2023-04-11，
  
  修回：2023-09-14，
  
  纸质出版：2024-02-15
- 稿件说明：
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赵光强, 南栋琪, 袁思琪, 等. 牛磺熊去氧胆酸通过抑制内质网应激诱导的凋亡促进类鼻疽菌的清除[J]. 陆军军医大学学报, 2024,46(3):225-231.

Guangqiang ZHAO, Dongqi NAN, Siqi YUAN, et al. TUDCA promotes intracellular clearance of Burkholderia pseudomallei by inhibiting endoplasmic reticulum stress-induced apoptosis in RAW264.7 cells[J]. Journal of Army Medical University, 2024, 46(3): 225-231.
赵光强, 南栋琪, 袁思琪, 等. 牛磺熊去氧胆酸通过抑制内质网应激诱导的凋亡促进类鼻疽菌的清除[J]. 陆军军医大学学报, 2024,46(3):225-231. DOI： 10.16016/j.2097-0927.202304046.

Guangqiang ZHAO, Dongqi NAN, Siqi YUAN, et al. TUDCA promotes intracellular clearance of Burkholderia pseudomallei by inhibiting endoplasmic reticulum stress-induced apoptosis in RAW264.7 cells[J]. Journal of Army Medical University, 2024, 46(3): 225-231. DOI： 10.16016/j.2097-0927.202304046.

摘要

目的

拟通过类鼻疽菌感染RAW264.7巨噬细胞模型，探讨牛磺熊去氧胆酸(tauroursodeoxycholic acid，TUDCA)促进类鼻疽菌胞内清除的作用机制。

方法

类鼻疽菌感染RAW264.7细胞后8 h，设置未感染组和类鼻疽菌感染组，在不同浓度TUDCA处理条件下，采用流式细胞仪检测类鼻疽菌诱导细胞凋亡的情况；同时使用另外一种内质网应激抑制剂4-PBA，通过Annexin-V/PI双染法和MTT细胞增殖实验分别检测类鼻疽菌感染对宿主细胞凋亡和增殖的影响；进一步使用siRNA干扰内质网应激关键基因CHOP，设置siC对照组和siCHOP干扰组，通过Western blot和流式细胞仪检测类鼻疽菌感染后Caspase-3、Caspase-12等细胞凋亡相关蛋白的表达水平及细胞凋亡情况；通过细菌胞内生存实验分析TUDCA通过抑制内质网应激减少其诱导的细胞凋亡、增强巨噬细胞对类鼻疽菌胞内清除作用的机制。

结果

在不同浓度TUDCA处理下，100 μmol/L或200 μmol/L TUDCA可明显减少类鼻疽菌感染诱导的RAW264.7细胞凋亡数量(

0.05)。分别使用两种内质网应激抑制剂TUDCA和4-PBA均可降低类鼻疽菌感染诱导的细胞凋亡(

0.05)。在siC对照组，相较于未感染时类鼻疽菌感染后Caspase-3和Caspase-12表达明显增加。在siCHOP干扰组，细胞凋亡相关蛋白的表达显著低于siC对照组，TUDCA处理后细胞凋亡相关蛋白表达水平与未加TUDCA药物相比无明显差异。流式细胞仪检测结果也显示，TUDCA能够减少类鼻疽菌感染后诱导的细胞凋亡(

0.05)，但在siCHOP干扰组中，TUDCA对类鼻疽菌感染诱导的凋亡无明显改变。细菌胞内生存计数结果显示，在siC对照组加入TUDCA能够增加宿主细胞对类鼻疽菌的清除(

0.05)，而在siCHOP干扰组中TUDCA的处理对类鼻疽菌胞内增殖无明显影响。

结论

在类鼻疽菌感染RAW264.7细胞模型中，TUDCA能够通过抑制巨噬细胞内质网应激诱导的凋亡，促进巨噬细胞对类鼻疽菌的胞内清除。

Abstract

Objective

To explore the action mechanism of tauroursodeoxycholic acid (TUDCA) promoting intracellular clearance of

Burkholderia pseudomallei

(

B. pseudomallei

) in RAW264.7 macrophages.

Methods

After TUDCA of different concentrations were used to treat RAW264.7 cells pre-infected with

B. pseudomallei

for 8 h or not

flow cytometry was applied to detect the apoptosis of the infected and control cells. In addition

another endoplasmic reticulum stress (ERS) inhibitor 4-PBA was used to detect the apoptosis and proliferation of host cells after

B. pseudomallei

infection with Annexin-V/PI double staining and MTT cell proliferation assay. Furthermore

after transfected with CHOP siRNA

Western blotting and flow cytometry were employed to detect the effect of TUDCA on the expression levels of Caspase-3 and Caspase-12 and the changes in apoptotic rate after

B. pseudomallei

infection

respectively. Finally

the effect of TUDCA on intracellular multiplication of infected RAW264.7 cells were observed to estimate the CFU value in the presence and absence of CHOP siRNA.

Results

Under different concentrations of TUDCA

100 or 200 μmol/L TUDCA significantly

reduced

B. pseudomallei

-induced apoptosis in RAW264.7 cells (

0.05). Meanwhile

both TUDCA and 4-PBA treatment could decrease the apoptosis induced by

B. pseudomallei

infection by ERS (

0.05). Further

the expression levels of Caspase-3 and Caspase-12 were obviously increased after

B. pseudomallei

infection compared with uninfected groups

but their expression levels in the siCHOP group was significantly lower than that in the siC group. Besides

flow cytometry also showed that TUDCA could reduce apoptosis induced by

B. pseudomallei

infection (

0.05)

but no significant effect of TUDCA on apoptosis was observed under CHOP knockdown. Finally

intracellular CFU assay indicated that TUDCA treatment promoted the host cell clearance of

B. pseudomallei

(

0.05)

but no such effect was observed in siCHOP group.

Conclusion

B. pseudomallei

infected RAW264.7 cells

TUDCA promotes the intracellular clearance of the bacteria by inhibiting ERS-induced apoptosis.

关键词

Keywords

references

WIERSINGA W J, VIRK H S, TORRES A G, et al . Melioidosis[J ] . Nat Rev Dis Primers, 2018, 4(1): 1-22. DOI: 10.1038/nrdp.2017.107.

BIRNIE E, VIRK H S, SAVELKOEL J, et al . Global burden of melioidosis in 2015: a systematic review and data synthesis[J ] . Lancet Infect Dis, 2019, 19(8): 892-902.DOI: 10.1016/S1473-3099(19)30157-4.

VIRK H S, MUKHOPADHYAY C, WIERSINGA W J. Melioidosis: a neglected cause of community-acquired pneumonia[J]. Semin Respir Crit Care Med, 2020, 41(4): 496-508. DOI: 10.1055/s-0040-1710570.

ROTZ L D, KHAN A S, LILLIBRIDGE S R, et al . Public health assessment of potential biological terrorism agents[J ] . Emerg Infect Dis, 2002, 8(2): 225-230. DOI: 10.3201/eid0802.010164.

毛旭虎, 林容, 方瑶, 等. 类鼻疽诊断与治疗专家共识[J]. 中华传染病杂志, 2022. 40(10): 577-583.

MAO X H, LIN R, FANG Y, et al . Expert consensus on diagnosis and treatment of melioidosis[J ] . Chin J Infect Dis, 2022. 40(10): 577-583.

CHOVATIYA R, MEDZHITOV R. Stress, inflammation, and defense of homeostasis[J]. Mol Cell, 2014, 54(2): 281-288. DOI: 10.1016/j.molcel.2014.03.030.

HEUBI J E, WIECHMANN D A, CREUTZINGER V, et al . Tauroursodeoxycholic acid (TUDCA) in the prevention of total parenteral nutrition-associated liver disease[J ] . J Pediatr, 2002, 141(2): 237-242. DOI: 10.1067/mpd.2002.125802.

OZCAN U, YILMAZ E, OZCAN L, et al . Chemical chaperones reduce ER stress and rest ore glucose homeostasis in a mouse model of type 2 diabetes[J ] . Science, 2006, 313(5790): 1137-1140. DOI: 10.1126/science.1128294.

DE MIGUEL C, SEDAKA R, KASZTAN M, et al . Tauroursodeoxycholic acid (TUDCA) abolishes chronic high salt-induced renal injury and inflammation[J ] . Acta Physiol (Oxf), 2019, 226(1): e13227. DOI: 10.1111/apha.13227.

ELIA A E, LALLI S, MONSURRÒ M R, et al . Tauroursodeoxycholic acid in the treatment of patients with amyotrophic lateral sclerosis[J ] . Eur J Neurol, 2016, 23(1): 45-52. DOI: 10.1111/ene.12664.

GASSIEP I, ARMSTRONG M, NORTON R. Human melioidosis[J]. Clin Microbiol Rev, 2020, 33(2): e00006-e00019. DOI: 10.1128/CMR.00006-19.

ALLWOOD E M, DEVENISH R J, PRESCOTT M, et al . Strategies for intracellular survival of Burkholderia pseudomallei [J ] . Front Microbio, 2011, 2: 170. DOI: 10.3389/fmicb.2011.00170..

WIERSINGA W J, CURRIE B J, PEACOCK S J. Melioidosis[J]. N Engl J Med, 2012, 367(11): 1035-1044. DOI: 10.1056/nejmra1204699.

LEE A S. The ER chaperone and signaling regulator GRP78/BiP as a monitor of endoplasmic reticulum stress[J]. Methods, 2005, 35(4): 373-381. DOI: 10.1016/j.ymeth.2004.10.010.

HILLARY R F, FITZGERALD U. A lifetime of stress: ATF6 in development and homeostasis[J]. J Biomed Sci, 2018, 25(1): 48. DOI: 10.1186/s12929-018-0453-1.

CELLI J, TSOLIS R. Bacteria, the endoplasmic reticulum and the unfolded protein response: friends or foes?[J]. Nat Rev Microbiol, 2014, 13: 71-82. DOI: 10.1038/nrmicro3393.

KAGAN J C, ROY C R. Legionella phagosomes intercept vesicular traffic from endoplasmic reticulum exit sites[J ] . Nat Cell Biol, 2002, 4(12): 945-954. DOI: 10.1038/ncb883.

ROBINSON C G, ROY C R. Attachment and fusion of endoplasmic reticulum with vacuoles containing Legionella pneumophila [J ] . Cell Microbiol, 2006, 8(5): 793-805. DOI: 10.1111/j.1462-5822.2005.00666.x.

SMITH J A, KHAN M, MAGNANI D D, et al . Brucella induces an unfolded protein response via TcpB tha t supports intracellular replication in macrophages[J ] . PLoS Pathog, 2013, 9(12): e1003785. DOI: 10.1371/journal.ppat.1003785..

LIM Y J, CHOI J A, CHOI H H, et al . Endoplasmic reticulum stress pathway-mediated apoptosis in macrophages contributes to the survival of Mycobacterium tuberculosis [J ] . PLoS One, 2011, 6(12): e28531. DOI: 10.1371/journal.pone.0028531..

LEE S Y, LEE M S, CHERLA R P, et al . Shiga toxin 1 induces apoptosis through the endoplasmic reticulum stress response in human monocytic cells[J ] . Cell Microbiol, 2008, 10(3): 770-780. DOI: 10.1111/j.1462-5822.2007.01083.x.

CHO J, LEE A H, PLATZER B, et al . RETRACTED: the unfolded protein response element IRE1α senses bacterial proteins invading the ER to activate RIG-I and innate immune signaling[J ] . Cell Host Microbe, 2013, 13(5): 558-569. DOI: 10.1016/j.chom.2013.03.011.

SENGUPTA D, KOBLANSKY A, GAINES J, et al . Subversion of innate immune responses by Brucella through the targeted degradation of the TLR signaling adapter, MAL[J ] . J Immunol, 2010, 184(2): 956-964. DOI: 10.4049/jimmunol.0902008..

SNYDER G A, DEREDGE D, WALDHUBER A, et al . Crystal structures of the Toll/interleukin-1 receptor (TIR) domains from the Brucella protein TcpB and host adaptor TIRAP reveal mechanisms of molecular mimicry[J ] . J Biol Chem, 2014, 289(2): 669-679. DOI: 10.1074/jbc.M113.523407..

SEIMON T A, KIM M J, BLUMENTHAL A, et al . Induction of ER stress in macrophages of tuberculosis granulomas[J ] . PLoS One, 2010. 5(9): e12772. DOI: 10.1371/journal.pone.0012772.

PILLICH H, CHAKRABORTY T, MRAHEIL M A. Cell-autonomous responses in Listeria monocytogenes infection[J ] . Future Microbiol, 2015, 10(4): 583-597. DOI: 10.2217/fmb.15.4.

MIMURA N, FULCINTI M, GORGUN G, et al . Blockade of XBP1 splicing by inhibition of IRE1alpha is a promising therapeutic option in multiple myeloma[J ] . Blood, 2012, 119(24): 5772-5781. DOI: 10.1182/blood-2011-07-366633.

YAN H, PENG B, LIU Y, et al . Viral entry of hepatitis B and D viruses and bile salts transportation share common molecular determinants on sodium taurocholate cotransporting polypeptide[J ] . J Virol, 2014, 88(6): 3273-3284. DOI: 10.1128/JVI.03478-13.

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