Trichostatin A improves the inflammatory response and liver injury in septic mice through the FoxO3a/autophagy signaling pathway

doi:10.5847/wjem.j.1920-8642.2022.056

Abstract

Abstract:

BACKGROUND: Sepsis-induced liver injury is a fatal complication of sepsis. Trichostatin A (TSA) regulates inflammation and autophagy in some human diseases, and forkhead box O3a (FoxO3a) has been shown to regulate autophagy. The present study aims to investigate whether TSA exerts its effects on septic liver injury through the FoxO3a/autophagy signaling pathway.

METHODS: A sepsis mouse model was constructed by the cecal ligation and puncture (CLP) method, and AML12 cells were pretreated with lipopolysaccharide (LPS) (1 μg/mL) to establish a sepsis cell model. Forty mice were divided into four groups, namely control group, TSA group, CLP group, and CLP+TSA group, with 10 mice in each group. Cells were divided into control group, TSA group, LPS group, and LPS+TSA group. Hematoxylin-eosin (H&E) staining and biochemical methods were used to evaluate liver tissue injury. Enzyme-linked immunosorbent assay (ELISA) was applied to detect the expression of proinflammatory cytokines, and Western blotting and immunofluorescence were used to measure autophagy-related protein expression.

RESULTS: Compared with the CLP group (mice), the proinflammatory cytokines (interleukin-β [IL-β] 2,665.27±324.90 pg/mL to 2,080.26±373.66 pg/mL; interleukin-6 [IL-6] 399.01±60.98 pg/mL to 221.90±46.89 pg/mL) and the hepatocyte injury markers (aspartate transaminase [AST] from 198.18±27.07 U/L to 128.42±20.55 U/L; alanine aminotransferase [ALT] from 634.98±74.10 U/L to 478.60±32.56 U/L) were notably decreased after TSA intervention. Moreover, LC3 II and FoxO3a showed an obvious increase and P62 showed an obvious decrease in the CLP+TSA group. Cell experiment results showed the similar trend. After FoxO3a gene was knocked down in AML12 cells, the promotion of autophagy and the improvement of liver enzyme index and inflammation by TSA were weakened.

CONCLUSION: TSA may improve the inflammatory response and liver injury in septic mice through FoxO3a/autophagy.

Key words: Sepsis, Liver injury, Trichostatin A, Autophagy, Inflammation

Mei-jia Shen, Li-chao Sun, Xiao-yu Liu, Meng-chen Xiong, Shan Li, A-ling Tang, Guo-qiang Zhang. Trichostatin A improves the inflammatory response and liver injury in septic mice through the FoxO3a/autophagy signaling pathway[J]. World Journal of Emergency Medicine, 2022, 13(3): 182-188.

Figures/Tables 5

Figure 1.

Table 1.

Figure 2.

Table 2.

Table 3.

References 26

1	Machado FR, Mazza BF. Improving mortality in sepsis: analysis of clinical trials. Shock. 2010; 34(Suppl 1):54-8. doi: 10.1097/SHK.0b013e3181e7e8b4
2	Mann EA, Baun MM, Meininger JC, Wade CE. Comparison of mortality associated with sepsis in the burn, trauma, and general intensive care unit patient: a systematic review of the literature. Shock. 2012; 37(1):4-16. doi: 10.1097/SHK.0b013e318237d6bf
3	Mizushima N, Ohsumi Y, Yoshimori T. Autophagosome formation in mammalian cells. Cell Struct Funct. 2002; 27(6):421-9. doi: 10.1247/csf.27.421 pmid: 12576635
4	Levine B, Mizushima N, Virgin HW. Autophagy in immunity and inflammation. Nature. 2011; 469(7330):323-35. doi: 10.1038/nature09782
5	Levine B, Kroemer G. Autophagy in the pathogenesis of disease. Cell. 2008; 132(1):27-42. doi: 10.1016/j.cell.2007.12.018 pmid: 18191218
6	Mizushima N, Levine B. Autophagy in mammalian development and differentiation. Nat Cell Biol. 2010; 12(9):823-30. doi: 10.1038/ncb0910-823 pmid: 20811354
7	Hsiao HW, Tsai KL, Wang LF, Chen YH, Chiang PC, Chuang SM, et al. The decline of autophagy contributes to proximal tubular dysfunction during sepsis. Shock. 2012; 37(3):289-96. doi: 10.1097/SHK.0b013e318240b52a
8	Shao L, Xiong X, Zhang Y, Miao H, Ren Y, Tang X, et al. IL-22 ameliorates LPS-induced acute liver injury by autophagy activation through ATF4-ATG7 signaling. Cell Death Dis. 2020; 11(11):970. doi: 10.1038/s41419-020-03176-4
9	Pei L, He L. Hepatoprotective effect of anemoside B4 against sepsis-induced acute liver injury through modulating the mTOR/p70S6K-mediated autophagy. Chem Biol Interact. 2021; 345:109534. doi: 10.1016/j.cbi.2021.109534
10	Yu Q, Zou L, Yuan X, Fang F, Xu F. Dexmedetomidine protects against septic liver injury by enhancing autophagy through activation of the AMPK/SIRT1 signaling pathway. Front Pharmacol. 2021; 12:658677. doi: 10.3389/fphar.2021.658677
11	Zhang JB, Ng S, Wang JG, Zhou J, Tan SH, Yang ND, et al. Histone deacetylase inhibitors induce autophagy through FoxO1-dependent pathways. Autophagy. 2015; 11(4):629-42. doi: 10.1080/15548627.2015.1023981
12	Kim SJ, Park JS, Lee DW, Lee SM. Trichostatin A protects liver against septic injury through inhibiting toll-like receptor signaling. Biomol Ther (Seoul). 2016; 24(4):387-94. doi: 10.4062/biomolther.2015.176
13	Cui SN, Chen ZY, Yang XB, Chen L, Yang YY, Pan SW, et al. Trichostatin A modulates the macrophage phenotype by enhancing autophagy to reduce inflammation during polymicrobial sepsis. Int Immunopharmacol. 2019; 77:105973. doi: 10.1016/j.intimp.2019.105973
14	Guo Y, Li Z, Shi C, Li J, Yao M, Chen X. Trichostatin A attenuates oxidative stress-mediated myocardial injury through the FoxO3a signaling pathway. Int J Mol Med. 2017; 40(4):999-1008. doi: 10.3892/ijmm.2017.3101
15	Liu YM, Lv J, Zeng QL, Shen S, Xing JY, Zhang YY, et al. AMPK activation ameliorates D-GalN/LPS-induced acute liver failure by upregulating FoxO3a to induce autophagy. Exp Cell Res. 2017; 358(2):335-42. doi: 10.1016/j.yexcr.2017.07.008
16	Funk DJ, Parrillo JE, Kumar A. Sepsis and septic shock: a history. Crit Care Clin. 2009; 25(1):83-101, viii. doi: 10.1016/j.ccc.2008.12.003
17	Deitch EA. Rodent models of intra-abdominal infection. Shock. 2005; 24(Suppl 1):19-23. doi: 10.1097/01.shk.0000191386.18818.0a
18	Gilmore TD. Introduction to NF-kappaB: players, pathways, perspectives. Oncogene. 2006; 25(51):6680-4. doi: 10.1038/sj.onc.1209954 pmid: 17072321
19	Takeuchi O, Akira S. Pattern recognition receptors and inflammation. Cell. 2010; 140(6):805-20. doi: 10.1016/j.cell.2010.01.022 pmid: 20303872
20	Takano KI, Yamamoto S, Tomita K, Takashina M, Yokoo H, Matsuda N, et al. Successful treatment of acute lung injury with pitavastatin in septic mice: potential role of glucocorticoid receptor expression in alveolar macrophages. J Pharmacol Exp Ther. 2011; 336(2):381-90. doi: 10.1124/jpet.110.171462
21	Oh JE, Lee HK. Pattern recognition receptors and autophagy. Front Immunol. 2014; 5:300.
22	Brealey D, Singer M. Mitochondrial dysfunction in Sepsis. Curr Infect Dis Rep. 2003; 5(5):365-71. doi: 10.1007/s11908-003-0015-9 pmid: 13678565
23	Shi CS, Shenderov K, Huang NN, Kabat J, Abu-Asab M, Fitzgerald KA, et al. Activation of autophagy by inflammatory signals limits IL-1β production by targeting ubiquitinated inflammasomes for destruction. Nat Immunol. 2012; 13(3):255-63. doi: 10.1038/ni.2215
24	Guo Y, Li Z, Shi C, Li J, Yao M, Chen X. Trichostatin A attenuates oxidative stress-mediated myocardial injury through the FoxO3a signaling pathway. Int J Mol Med. 2017; 40(4):999-1008. doi: 10.3892/ijmm.2017.3101
25	Ali T, Rahman SU, Hao Q, Li W, Liu Z, Ali Shah F, et al. Melatonin prevents neuroinflammation and relieves depression by attenuating autophagy impairment through FoxO3a regulation. J Pineal Res. 2020; 69(2):e12667.
26	Wu Y, Leng Y, Meng Q, Xue R, Zhao B, Zhan L, et al. Suppression of excessive histone deacetylases activity in diabetic hearts attenuates myocardial ischemia/reperfusion injury via mitochondria apoptosis pathway. J Diabetes Res. 2017; 2017: 8208065.

Parameters	Control group	TSA group	CLP group	CLP+TSA group
ALT ^a, U/L	54.92±9.68	34.36±12.90	198.18±27.07^**	128.42±20.55^#
AST ^a, U/L	206.05±17.11	100.49±6.39^**	634.98±74.10^**	478.60±32.56^#
IL-6 ^a, pg/mL	371.62±27.69	306.04±13.05^**	2,665.27±324.90^**	2,080.26±373.66^#
TNF-α ^a, pg/mL	177.79±24.72	135.00±17.40^*	399.01±60.98^**	221.90±46.89^##
LC3 II ^b	1	1.46±0.02^**	1.36±0.01^**	1.66±0.04^##
P62 ^b	1	0.60±0.02^**	0.87±0.02^**	0.59±0.02^##
FoxO3a ^b	1	2.03±0.13^**	1.49±0.02^**	2.89±0.03^##

Parameters	Control group	TSA group	LPS group	LPS+TSA group
ALT ^a, U/L	5.05±0.55	14.20±2.20	29.42±0.62^**	23.47±0.64^##
AST ^a, U/L	9.55±0.65	18.90±3.10	63.52±4.50^**	38.51±1.50^##
LC3 II ^b	1	1.77±0.02^**	1.39±0.01^**	1.87±0.01^##
P62 ^b	1	0.50±0.01^**	0.73±0.01^**	0.38±0.01^##
Foxo3a ^b	1	2.23±0.02^**	1.79±0.01^**	3.10±0.07^##

Parameters	Control group	si-negative control			si-FoxO3a
Parameters	Control group	TSA group	LPS group	LPS+TSA group	TSA group	LPS group	LPS+TSA group
ALT ^a, U/L	5.37±0.63	14.13±1.80	29.27±0.52	23.33±0.50	14.63±1.03	38.57±2.30^**	29.20±1.10^**
AST ^a, U/L	9.07±0.87	20.27±3.18	57.00±1.59	39.57±1.94	19.63±3.03	71.23±5.85^*	46.70±2.90^*
IL-6 ^a, pg/mL	40.05±1.96	111.77±2.53	500.50±7.96	355.07±9.71	132.07±2.76^**	598.57±14.01^**	413.73±26.24^*
TNF-α ^a, pg/mL	10.33±1.25	11.93±1.69	54.15±2.79	36.77±2.39	13.47±1.92^*	66.47±3.03^*	50.43±3.35^**
LC3 II^b	1	1.51±0.01	2.51±0.02	2.57±0.01	1.22±0.01^**	1.69±0.01^**	1.28±0.04^**
P62 ^b	1	0.48±0.01	0.54±0.01	0.33±0.01	0.94±0.01^**	0.66±0.01^**	0.93±0.02^**

[1]	Xuan Fu, Xue Lin, Samuel Seery, Li-na Zhao, Hua-dong Zhu, Jun Xu, Xue-zhong Yu. Speckle-tracking echocardiography for detecting myocardial dysfunction in sepsis and septic shock patients: A single emergency department study [J]. World Journal of Emergency Medicine, 2022, 13(3): 175-181.
[2]	Hai Hu, Jing-yuan Jiang, Ni Yao. Comparison of different versions of the quick sequential organ failure assessment for predicting in-hospital mortality of sepsis patients: A retrospective observational study [J]. World Journal of Emergency Medicine, 2022, 13(2): 114-119.
[3]	Li-wei Duan, Jin-long Qu, Jian Wan, Yong-hua Xu, Yi Shan, Li-xue Wu, Jin-hao Zheng, Wei-wei Jiang, Qi-tong Chen, Yan Zhu, Jian Zhou, Wen-bo Yu, Lei Pei, Xi Song, Wen-fang Li, Zhao-fen Lin. Effects of viral infection and microbial diversity on patients with sepsis: A retrospective study based on metagenomic next-generation sequencing [J]. World Journal of Emergency Medicine, 2021, 12(1): 29-35.
[4]	Hai-jiang Zhou, Tian-fei Lan, Shu-bin Guo. Outcome prediction value of National Early Warning Score in septic patients with community-acquired pneumonia in emergency department: A single-center retrospective cohort study [J]. World Journal of Emergency Medicine, 2020, 11(4): 206-215.
[5]	Yu-ming Wang, Yan-jun Zheng, Ying Chen, Yun-chuan Huang, Wei-wei Chen, Ran Ji, Li-li Xu, Zhi-tao Yang, Hui-qiu Sheng, Hong-ping Qu, En-qiang Mao, Er-zhen Chen. Effects of fluid balance on prognosis of acute respiratory distress syndrome patients secondary to sepsis [J]. World Journal of Emergency Medicine, 2020, 11(4): 216-222.
[6]	Miao Yuan, Ding-yi Yan, Fang-shi Xu, Yi-di Zhao, Yang Zhou, Long-fei Pan. Effects of sepsis on hippocampal volume and memory function [J]. World Journal of Emergency Medicine, 2020, 11(4): 223-230.
[7]	Wen-peng Yin, Jia-bao Li, Xiao-fang Zheng, Le An, Huan Shao, Chun-sheng Li. Effect of neutrophil CD64 for diagnosing sepsis in emergency department [J]. World Journal of Emergency Medicine, 2020, 11(2): 79-86.
[8]	Shao-hua Liu, Huo-yan Liang, Hong-yi Li, Xian-fei Ding, Tong-wen Sun, Jing Wang. Effect of low high-density lipoprotein levels on mortality of septic patients: A systematic review and meta-analysis of cohort studies [J]. World Journal of Emergency Medicine, 2020, 11(2): 109-116.
[9]	Yi-wen Fan, Shao-wei Jiang, Jia-meng Chen, Hui-qi Wang, Dan Liu, Shu-ming Pan, Cheng-jin Gao. A pulmonary source of infection in patients with sepsis-associated acute kidney injury leads to a worse outcome and poor recovery of kidney function [J]. World Journal of Emergency Medicine, 2020, 11(1): 18-26.
[10]	Kimberly A. Chambers, Adam Y. Park, Rosa C. Banuelos, Bryan F. Darger, Bindu H. Akkanti, Annamaria Macaluso, Manoj Thangam, Pratik B. Doshi. Outcomes of severe sepsis and septic shock patients after stratification by initial lactate value [J]. World Journal of Emergency Medicine, 2018, 9(2): 113-117.
[11]	Muhammad Akbar Baig, Hira Shahzad, Erfan Hussain, Asad Mian. Validating a point of care lactate meter in adult patients with sepsis presenting to the emergency department of a tertiary care hospital of a low- to middle-income country [J]. World Journal of Emergency Medicine, 2017, 8(3): 184-189.
[12]	Ruo Wu, Luo-gen Peng, Hui-min Zhao. Diverse coagulopathies in a rabbit model with different abdominal injuries [J]. World Journal of Emergency Medicine, 2017, 8(2): 141-147.
[13]	Jia-jun Xu, Jian-tao Zhen, Li Tang, Qing-ming Lin. Intravenous injection of Xuebijing attenuates acute kidney injury in rats with paraquat intoxication [J]. World Journal of Emergency Medicine, 2017, 8(1): 61-64.
[14]	Chao Cao, Tao Ma, Yan-fen Chai, Song-tao Shou. The role of regulatory T cells in immune dysfunction during sepsis [J]. World Journal of Emergency Medicine, 2015, 6(1): 5-9.
[15]	Wei-ping Sun, Guang-xiong Yuan, Yan-juan Hu, Li-zhen Liao, Lin Fu. Effect of low-dose glucocorticoid on corticosteroid insufficient patients with acute exacerbation of chronic obstructive pulmonary disease [J]. World Journal of Emergency Medicine, 2015, 6(1): 34-39.