Sodium dichloroacetate improves migration ability by suppressing LPS-induced inflammation in HTR-8/SVneo cells via the TLR4/NF-κB pathway

Document Type : Original Article

Authors

1 School of Public Health and Hongqiao International Institute of Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

2 Department of Pharmacy, Second Affiliated Hospital of Naval Medical University, Shanghai 200003, China

3 Department of Biochemical Pharmacy, School of Pharmacy, Naval Medical University, Shanghai 200433, China

4 School of Medicine, Shanghai University, Shanghai 200444, China

5 Institute for Developmental and Regenerative Cardiovascular Medicine, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China

Abstract

Objective(s): Inadequate cytotrophoblast migration and invasion are speculated to result in preeclampsia, which is a pro-inflammatory condition. Sodium dichloroacetate (DCA) exerts anti-inflammatory actions. Thus,we sought to investigate the effect of DCA on the migration function of the lipopolysaccharide (LPS)-stimulated human-trophoblast-derived cell line (HTR-8/SVneo).
Materials and Methods: HTR-8/SVneo cells were treated with LPS to suppress cell migration. Cell migration was examined by both scratch wound healing assay and transwell migration assay. Western blotting was used to analyze the expression levels of toll-like receptor-4 (TLR4), nuclear factor-κB (NF-κB), TNF-α, IL-1β, and IL-6 in the cells.
Results: DCA reversed LPS-induced inhibition of migration in HTR-8/SVneo cells. Furthermore, DCA significantly suppressed LPS-induced activation of TLR4, phosphorylation of NF-κB (p65), translocation of p65 into the nucleus, and the production of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6). Treatment with inhibitors of TLR4 signal transduction (CLI095 or MD2-TLR-4-IN-1) reduced LPS-induced overexpression of pro-inflammatory cytokines, and a synergistic effect was found between TLR4 inhibitors and DCA in HTR-8/SVneo cells. 
Conclusion: DCA improved trophoblast cell migration function by suppressing LPS-induced inflammation, at least in part, via the TLR4/NF-κB signaling pathway. This result indicates that DCA might be a potential therapeutic candidate for human pregnancy-related complications associated with trophoblast disorder.

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Main Subjects


1. Amaral LM, Cornelius DC, Harmon A, Moseley J, Martin JN Jr, LaMarca B. 17-hydroxyprogesterone caproate significantly improves clinical characteristics of PE in the reduced uterine perfusion pressure rat model. Hypertension 2015; 65: 225-231.
2.    Melchiorre K, Sharma R, Thilaganathan B. Cardiovascular implications in preeclampsia: An overview. Circulation 2014; 130: 703-714.
3.    Say L, Chou D, Gemmill A, Tunçalp Ö, Moller AB, Daniels J, Gülmezoglu AM, Temmerman M, Alkema L. Global causes of maternal death: A WHO systematic analysis. Lancet Glob Health 2014; 2: e323-333. 
4.    Redman CW, Sacks GP, Sargent IL. Preeclampsia: An excessive maternal inflammatory response to pregnancy. Am J Obstet Gynecol 1999; 180: 499-506.
5.    Damsky CH, Librach C, Lim KH, Fitzgerald ML, McMaster MT, Janatpour M, et al. Integrin switching regulates normal trophoblast invasion. Development 1994; 120: 3657-3666.
6.    Kliman HJ, Nestler JE, Sermasi E, Sanger JM, Strauss JF 3rd. Purification, characterization, and in vitro differentiation of cytotrophoblasts from human term placentae. Endocrinology 1986; 118: 1567-1582. 
7.    Kohli S, Ranjan S, Hoffmann J, Kashif M, Daniel EA, Al-Dabet MM et al. Maternal extracellular vesicles and platelets promote preeclampsia via inflammasome activation in trophoblasts. Blood 2016; 128: 2153-2164. 
8.    Noris M, Perico N, Remuzzi G. Mechanisms of disease: Pre-eclampsia. Nat Clin Pract Nephrol 2005; 1: 98-114.
9.    Redman CW, Sargent IL. Latest advances in understanding preeclampsia. Science 2005; 308: 1592-1594.
10.    Svinarich DM, Bitonti OM, Romero R, Gonik B. Induction and posttranslational expression of cytokines in a first-trimester trophoblast cell line by lipopolysaccharide. Am J Obstet Gynecol 1996; 175: 970-973. 
11.    Zaga-Clavellina V, Garcia-Lopez G, Flores-Herrera H, Espejel-Nuñez A, Flores-Pliego A, Soriano-Becerril et al. In vitro secretion profiles of interleukin (IL)-1beta, IL-6, IL-8, IL-10, and TNF alpha after selective infection with Escherichia coli in human fetal membranes. Reprod Biol Endocrinol 2007; 5: 1-7.
12.    Dowling O, Chatterjee PK, Gupta M, Tam Tam HB, Xue X, Lewis D et al. Magnesium sulfate reduces bacterial LPS-induced inflammation at the maternal-fetal interface. Placenta 2012; 33: 392-398.
13.    Stacpoole PW. The pharmacology of dichloroacetate. Metabolism 1989; 38: 1124-1144.
14.    Michelakis ED, Sutendra G, Dromparis P, Webster L, Haromy A, Niven E et al. Metabolic modulation of glioblastoma with dichloroacetate. Sci Transl Med 2010; 2: 31ra34. 
15.    Semba H, Takeda N, Isagawa T, Sugiura Y, Honda K, Wake M et al. HIF-1α-PDK1 axis-induced active glycolysis plays an essential role in macrophage migratory capacity. Nat Commun 2016; 7: 1-10. 
16.    Bian L, Josefsson E, Jonsson IM, Verdrengh M, Ohlsson C, Bokarewa M et al. Dichloroacetate alleviates development of collagen II-induced arthritis in female DBA/1 mice. Arthritis Res Ther 2009; 11: 1-10. 
17.    Horne AW, Ahmad SF, Carter R, Simitsidellis I, Greaves E, Hogg C et al. Repurposing dichloroacetate for the treatment of women with endometriosis. Proc Natl Acad Sci U S A 2019; 116: 25389-25391.
18.    Shakhov AN, Collart MA, Vassalli P, Nedospasov SA, Jongeneel CV. Kappa B-type enhancers are involved in lipopolysaccharide-mediated transcriptional activation of the tumor necrosis factor alpha gene in primary macrophages. J Exp Med 1990; 171: 35-47. 
19.    Landoni VI, Schierloh P, de Campos Nebel M, Fernández GC, Calatayud C, Lapponi MJ et al. Shiga toxin 1 induces on lipopolysaccharide-treated astrocytes the release of tumor necrosis factor-alpha that alter brain-like endothelium integrity. PLoS Pathog 2012; 8: e1002632. 
20.    Poltorak A, He X, Smirnova I, Liu MY, Van Huffel C, Du X et al. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 1998; 282: 2085-2088.
21.    Yang X, Haghiac M, Glazebrook P, Minium J, Catalano PM, Hauguel-de Mouzon S. Saturated fatty acids enhance TLR4 immune pathways in human trophoblasts. Hum Reprod. 2015; 30: 2152-2159. 
22.    Cramer M, Nagy I, Murphy BJ, Gassmann M, Hottiger MO, Georgiev O et al. NF-kappaB contributes to transcription of placenta growth factor and interacts with metal responsive transcription factor-1 in hypoxic human cells. Biol Chem 2005; 386: 865-872. 
23.    Logothetou-Rella H, Kotoulas IG, Nesland JM, Kipiotis D, Abazis D. Early human trophoblast cell cultures. A morphological and immunocytochemical study. Histol Histopathol 1989; 4: 367-374. 
24.    Chen YY, Xiao L, Sun GQ, Li M, Yang HL, Ming ZY et al. TMBIM4 deficiency facilitates NLRP3 inflammasome activation-induced pyroptosis of trophoblasts: A potential pathogenesis of preeclampsia. Biology (Basel) 2023; 12: 1-17.
25.    Liang CC, Park AY, Guan JL. In vitro scratch assay: A convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2007; 2: 329-333.
26.    Jonkman JEN, Cathcart JA, Xu F, Bartolini ME, Amon JE, Stevens KM et al. An introduction to the wound healing assay using live-cell microscopy. Cell Adh Migr 2014; 8: 440-451.
27.    Justus CR, Leffler N, Ruiz-Echevarria M, Yang LV. In vitro cell migration and invasion assays. J Vis Exp 2014; 88: 1-8.
28.    Abbas Y, Turco MY, Burton GJ, Moffett A. Investigation of human trophoblast invasion in vitro. Hum Reprod Update 2020; 26: 501-513.
29.    Peng C, Yang LJ, Zhang C, Jiang Y, Shang LLWX, He JB et al. Low-dose nifedipine rescues impaired endothelial progenitor cell-mediated angiogenesis in diabetic mice. Acta Pharmacol Sin 2023; 44: 44-57.
30.    Otsuka K, Terasaki F, Ikemoto M, Fujita SC, Tsukada B, Katashima T et al. Suppression of inflammation in rat autoimmune myocarditis by S100A8/A9 through modulation of the proinflammatory cytokine network. Eur J Heart Fail 2009; 11: 229-237.
31.    Liu KH, Zhang BL, Yang BS, Shi WT, Li YF, Wang Y et al. Rhizobiales-specific RirA represses a naturally “synthetic” foreign siderophore gene cluster to maintain sinorhizobium-legume mutualism. mBio 2021; 13: e0290021.
32.    Qi D, Lu JY, Fu ZY, Lv SS, Hou LL. Psoralen promotes proliferation, migration, and invasion of human extravillous trophoblast derived HTR-8/Svneo cells in vitro by NF-κB pathway. Front Pharmacol 2022; 13: 1-10.
33.    Janovec V, Ryabchenko B, Škarková A, Pokorná K, Rösel D, Brábek J et al. TLR4-mediated recognition of mouse polyomavirus promotes cancer-associated fibroblast-like phenotype and cell invasiveness. Cancers (Basel) 2021; 13: 1-16.
34.    Oh KK, Adnan M, Cho DH. Drug investigation to dampen the comorbidity of rheumatoid arthritis and osteoporosis via molecular docking test. Curr Issues Mol Biol 2022; 44: 1046-1061.
35.    Zhang YY, Zhang Y, Yao YB, Lei XL, Qian ZJ. Butyrolactone-I from coral-derived fungus aspergillus terreus attenuates neuro-inflammatory response via suppression of NF-κB pathway in BV-2 cells. Mar Drugs 2018; 16: 1-13. 
36.    Du M, Yuan L, Tan X, Huang D, Wang X, Zheng Z et al. The LPS-inducible lncRNA Mirt2 is a negative regulator of inflammation. Nat Commun 2017; 8: 1-18. 
37.    Gao H, Sun W, Zhao J, Wu X, Lu JJ, Chen X et al. Tanshinones and diethyl blechnics with anti-inflammatory and anti-cancer activities from Salvia miltiorrhiza Bunge (Danshen). Sci Rep 2016; 26: 1-13. 
38.    Askie LM, Duley L, Henderson-Smart DJ, Stewart LA. Antiplatelet agents for prevention of pre-eclampsia: A meta-analysis of individual patient data. Lancet 2007; 369: 1791-1798. 
39.    Davies JE, Pollheimer J, Yong HEJ, Kokkinos MI, Kalionis B, Knöfler M et al. Epithelial-mesenchymal transition during extravillous trophoblast differentiation. Cell Adh Migr 2016; 10: 310-321. 
40.    Msheik H, Azar J, Sabeh ME, Abou-Kheir W, Daoud G. HTR-8/SVneo: A model for epithelial to mesenchymal transition in the human placenta. Placenta 2020; 90: 90-97. 
41.    Abou-Kheir W, Barrak J, Hadadeh O, Daoud G. HTR-8/SVneo cell line contains a mixed population of cells. Placenta 2017; 50: 1-7. 
42.    Granger JP. Inflammatory cytokines, vascular function, and hypertension. Am J Physiol Regul Integr Comp Physiol 2004; 286: R989-R990.
43.    Takashima K, Matsunaga N, Yoshimatsu M, Hazeki K, Kaisho T, Uekata M et al. Analysis of binding site for the novel small-molecule TLR4 signal transduction inhibitor TAK-242 and its therapeutic effect on mouse sepsis model. Br J Pharmacol 2009; 157: 1250-1262. 
44.    Liu Z, Chen L, Yu P, Zhang Y, Fang B, Wu C et al. Discovery of 3-(Indol-5-yl)-indazole derivatives as novel myeloid differentiation protein 2/Toll-like receptor 4 antagonists for treatment of acute lung injury. J Med Chem 2019; 62: 5453-5469. 
45.    Shimazu R, Akashi S, Ogata H, Nagai Y, Fukudome K, Miyake K et al. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med 1999; 189: 1777-1782. 
46.    Vaughan JE, Walsh SW. Activation of NF-κB in placentas of women with preeclampsia. Hypertens Pregnancy 2012; 31: 243-251.