Sirt1 involved in sepsis-induced acute liver injury via regulating HNF-1α/FXR-1 pathway: The mechanism study in a rodent model

doi:10.16150/j.1671-2870.2020.03.014

Abstract

Abstract:

Objective: To investigate the role of Silent information regulator 1 (SIRT1) in sepsis-induced liver injury and the related mechanism in a mouse model. Methods: A mouse sepsis model was established by intraperitoneal injection (IP) of LPS, and the mice in control group were only given same volume sterilized 0.9% saline via IP. Sections of the liver from the mice in both groups were stained with hematoxylin-eosin and hepatic injury was examined, and quantitative real-time polymerase chain reaction (qrt-PCR) was performed to detect expression of Sirt1/HNF-1α/FXR-1 in hepatic tissue. The isolated primary mouse hepatic parenchymal cells were treated with different concentrations of LPS. Viabilityand proliferation of hepatic parenchymal cells were evaluated by CCK8 and EdU assay respectively. In addition, enzyme linked immunosorbent assay (ELISA) was conducted to measure interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) production in hepatic parenchymal cells treated with different concentration of LPS. Sirt1 overexpressing plasmids were constructed and transfected into hepatic cells in mediation of Lipofectamine 2000. After 48 hour, the Sirt1/HNF-1α/FXR-1 pathway expression was tested by qrt-PCR and the impact of overexpressing Sirt1 on the function of the cells was evaluated by CCK8, EdU and ELISA. Results: Compared with control, Sirt1/HNF-1α/FXR-1 pathway was down-regulated in LPS-induced sepsis mouse model. LPS lowered cell viability, inhibited proliferation and increased secretion of inflammatory factors IL-6 and TNF-α in hepatocytes. Over-expression of Sirt1 upregulated the expression of HNF-1α/FXR-1 pathway in parenchymal cells, alleviated LPS-induced inhibition of cell viability and proliferation, and decreased LPS-induced inflammatory factor secretion. Conclusions: The Sirt1/HNF-1α/FXR-1 pathway is involved in LPS-induced septic liver injury. Overexpression of Sirt1 is able to attenuate the toxic effects of LPS on parenchymal cells.

Key words: Sepsis, LPS, Silent information regulator 1 (SIRT1), HNF-1α/FXR-1 pathway

CLC Number:

R575.3

WANG Qiuyun, CHEN Ying, ZHAO Bing, SUN Silei, YANG Zhitao, MAO Enqiang, CHEN Erzhen. Sirt1 involved in sepsis-induced acute liver injury via regulating HNF-1α/FXR-1 pathway: The mechanism study in a rodent model[J]. Journal of Diagnostics Concepts & Practice, 2020, 19(03): 279-285.

Figures/Tables 6

References 21

[1]	Singer M, Deutschman CS, Seymour CW, et al. The third international consensus definitions for sepsis and septic shock (Sepsis-3)[J]. JAMA, 2016, 315(8):801-810. doi: 10.1001/jama.2016.0287 URL
[2]	Prescott HC, Angus DC. Postsepsis morbidity[J]. JAMA, 2018, 319(1):91. doi: 10.1001/jama.2017.19809 pmid: 29297079
[3]	Reinhart K, Daniels R, Kissoon N, et al. Recognizing sepsis as a global health priority-A WHO Resolution[J]. N Engl J Med, 2017, 377(5):414-417. doi: 10.1056/NEJMp1707170 URL
[4]	Levy MM, Evans LE, Rhodes A. The Surviving Sepsis Campaign Bundle: 2018 update[J]. Intensive Care Med, 2018, 44(6):925-928. doi: 10.1007/s00134-018-5085-0 URL
[5]	中国医师协会急诊医师分会, 中国研究型医院学会休克与脓毒症专业委员会. 中国脓毒症/脓毒性休克急诊治疗指南(2018)[J]. 临床急诊杂志, 2018, 19(9)423-444.
[6]	Kaffarnik MF, Lock JF, Vetter H, et al. Early diagnosis of sepsis-related hepatic dysfunction and its prognostic impact on survival: A prospective study with the LiMAx test[J]. Crit Care, 17(5):R259. doi: 10.1186/cc13089 URL
[7]	Zhao P, Wang C, Liu W, et al. Causes and outcomes of acute liver failure in China[J]. PLoS One, 2013, 8(11):e80991. doi: 10.1371/journal.pone.0080991 URL
[8]	Huang W, Ma K, Zhang J, et al. Nuclear receptor-dependent bile acid signaling is required for normal liver regeneration[J]. Science, 2006, 312(5771):233-236. doi: 10.1126/science.1121435 URL
[9]	Kemper JK, Xiao Z, Ponugoti B, et al. FXR acetylation is normally dynamically regulated by p300 and SIRT1 but constitutively elevated in metabolic disease states[J]. Cell Metab, 2009, 10(5):392-404. doi: 10.1016/j.cmet.2009.09.009 URL
[10]	Kazgan N, Metukuri MR, Purushotham A, et al. Intestine-specific deletion of SIRT1 in mice impairs DCoH2-HNF-1α-FXR signaling and alters systemic bile acid homeo-stasis[J]. Gastroenterology, 2014, 146(4):1006-1016. doi: 10.1053/j.gastro.2013.12.029 URL
[11]	Xie J, Zhang X, Zhang L. Negative regulation of inflammation by SIRT1[J]. Pharmacol Res, 2013, 67(1):60-67. doi: 10.1016/j.phrs.2012.10.010 URL
[12]	Tang BL. Sirt1 and the mitochondria[J]. Mol Cells, 2016, 39(2):87-95. doi: 10.14348/molcells.2016.2318 URL
[13]	Meng X, Tan J, Li M, et al. Sirt1: Role under the condition of ischemia/hypoxia[J]. Cell Mol Neurobiol, 2017, 37(1):17-28. doi: 10.1007/s10571-016-0355-2 URL
[14]	Nakamura K, Zhang M, Kageyama S, et al. Macrophage heme oxygenase-1-SIRT1-p53 axis regulates sterile inflammation in liver ischemia-reperfusion injury[J]. J Hepatol, 2017, 67(6):1232-1242. doi: S0168-8278(17)32217-1 pmid: 28842295
[15]	Zhu Y, Wang K, Ma Z, et al. SIRT1 activation by butein attenuates sepsis-induced brain injury in mice subjected to cecal ligation and puncture via alleviating inflammatory and oxidative stress[J]. Toxicol Appl Pharmacol, 2019, 363:34-46. doi: 10.1016/j.taap.2018.10.013 URL
[16]	Cui X, Chen Q, Dong Z, et al. Inactivation of Sirt1 in mouse livers protects against endotoxemic liver injury by acetylating and activating NF-κB[J]. Cell Death Dis, 2016, 7(10):e2403. doi: 10.1038/cddis.2016.270 URL
[17]	Zhuo Y, Zhang S, Li C, et al. Resolvin D1 promotes SIRT1 expression to counteract the activation of STAT3 and NF-κB in mice with septic-associated lung injury[J]. Inflammation, 2018, 41(5):1762-1771. doi: 10.1007/s10753-018-0819-2 URL
[18]	Lan KC, Chao SC, Wu HY, et al. Salidroside ameliorates sepsis-induced acute lung injury and mortality via downregulating NF-κB and HMGB1 pathways through the upregulation of SIRT1[J]. Sci Rep, 2017, 7(1):12026. doi: 10.1038/s41598-017-12285-8 URL
[19]	Hong G, Zheng D, Zhang L, et al. Administration of nicotinamide riboside prevents oxidative stress and organ injury in sepsis[J]. Free Radic Biol Med, 2018, 123:125-137. doi: 10.1016/j.freeradbiomed.2018.05.073 URL
[20]	Li K, Lv G, Pan L. Sirt1 alleviates LPS induced inflammation of periodontal ligament fibroblasts via downregulation of TLR4[J]. Int J Biol Macromol, 2018, 119:249-254. doi: 10.1016/j.ijbiomac.2018.07.099 URL
[21]	Purushotham A, Xu Q, Lu J, et al. Hepatic deletion of SIRT1 decreases hepatocyte nuclear factor 1α/farnesoid X receptor signaling and induces formation of cholesterol gallstones in mice[J]. Mol Cell Biol, 2012, 32(7):1226-1236. doi: 10.1128/MCB.05988-11 pmid: 22290433

基因	引物序列
Sirtq1上游	5'-GATACGGAGAGGCCCGAATG-3'
Sirtq1下游	5'-AGTTCCCAATGCTGGTGGAG-3'
FXR-1上游	5'-TCCTTGTCCTCCTCGGAACA-3'
FXR-1下游	5'-TGGGGTTTCCTGAAGCCTTG-3'
HNF-1α上游	5'-GTACGGACAGTCTGCAACCA-3'
HNF-1α下游	5'-TACTTGGTGTAAGGCCGCAG-3'
GAPDH上游	5'-AGGTCGGTGTGAACGGATTTG-3'
GAPDH下游	5'-GGGGTCGTTGATGGCAACA-3'