CTRP9 overexpression attenuates palmitic acid‑induced inflammation, apoptosis and impaired migration in HTR8/SVneo cells through AMPK/SREBP1c signaling

Li,Li; Gu,Zhongyi; Zhang,Junjie

doi:10.3892/etm.2022.11386

CTRP9 overexpression attenuates palmitic acid‑induced inflammation, apoptosis and impaired migration in HTR8/SVneo cells through AMPK/SREBP1c signaling

Authors:
- Li Li
- Zhongyi Gu
- Junjie Zhang
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Affiliations: Department of Obstetrics and Gynecology, Changhai Hospital, Naval Medical University, Shanghai 200433, P.R. China
Published online on: May 20, 2022 https://doi.org/10.3892/etm.2022.11386
Article Number: 459
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Obesity in pregnant mothers often leads to a range of obstetric complications, including miscarriage, pre‑eclampsia, gestational hypertension and diabetes. C1q/TNF‑related protein 9 (CTRP9) is an adipokine with an anti‑inflammatory effect. The aim of the present study was to identify the role of CTRP9 in the pathogenesis of maternal obesity during pregnancy. Following treatment with palmitic acid (PA), HTR8/SVneo cell viability and CTRP9 expression were analyzed using Cell Counting Kit‑8 (CCK‑8), reverse transcription‑quantitative PCR (RT‑qPCR) and western blot analyses. The effects of CTRP9 overexpression on cell viability, apoptosis, pro‑inflammatory cytokine levels and migration were assessed using CCK‑8, TUNEL, RT‑qPCR and Transwell assays, respectively. Subsequently, sterol‑regulatory element binding protein 1c (SREBP1c) overexpression efficiency was verified using RT‑qPCR, and its effects on cell viability, apoptosis, pro‑inflammatory cytokines and migration damage were then examined in HTR8/SVneo cells. The results showed that CTRP9 overexpression attenuated the inhibition of cell viability and apoptosis caused by PA in HTR8/SVneo cells, reduced pro‑inflammatory cytokine release, improved cell migration and regulated the protein expression level of AMP‑activated protein kinase (AMPK)/SREBP1c signaling. In addition, CTRP9 inhibited SREBP1c expression through AMPK signaling, thereby attenuating the inflammation, apoptosis and inhibited migration caused by PA in HTR8/SVneo cells. In brief, CTRP9 protected against inflammation, apoptosis and migration defects in HTR8/SVneo cells exposed to PA treatment through AMPK/SREBP1c signaling, which suggested the potential role of CTRP9 in alleviating the toxicity of PA.

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1	Stubert J, Reister F, Hartmann S and Janni W: The risks associated with obesity in pregnancy. Dtsch Arztebl Int. 115:276–283. 2018.PubMed/NCBI View Article : Google Scholar
2	Flegal KM, Carroll MD, Ogden CL and Curtin LR: Prevalence and trends in obesity among US adults, 1999-2008. JAMA. 303:235–241. 2010.PubMed/NCBI View Article : Google Scholar
3	Marchi J, Berg M, Dencker A, Olander EK and Begley C: Risks associated with obesity in pregnancy, for the mother and baby: A systematic review of reviews. Obes Rev. 16:621–638. 2015.PubMed/NCBI View Article : Google Scholar
4	Vasudevan C, Renfrew M and McGuire W: Fetal and perinatal consequences of maternal obesity. Arch Dis Child Fetal Neonatal Ed. 96:F378–F382. 2011.PubMed/NCBI View Article : Google Scholar
5	Basu S, Haghiac M, Surace P, Challier JC, Guerre-Millo M, Singh K, Waters T, Minium J, Presley L, Catalano PM, et al: Pregravid obesity associates with increased maternal endotoxemia and metabolic inflammation. Obesity (Silver Spring). 19:476–482. 2011.PubMed/NCBI View Article : Google Scholar
6	Yang X, Haghiac M, Glazebrook P, Minium J, Catalano PM and Hauguel-de Mouzon S: Saturated fatty acids enhance TLR4 immune pathways in human trophoblasts. Hum Reprod. 30:2152–2159. 2015.PubMed/NCBI View Article : Google Scholar
7	Pantham P, Aye IL and Powell TL: Inflammation in maternal obesity and gestational diabetes mellitus. Placenta. 36:709–715. 2015.PubMed/NCBI View Article : Google Scholar
8	Rampersaud AM, Dunk CE, Lye SJ and Renaud SJ: Palmitic acid induces inflammation in placental trophoblasts and impairs their migration toward smooth muscle cells through plasminogen activator inhibitor-1. Mol Hum Reprod. 26:850–865. 2020.PubMed/NCBI View Article : Google Scholar
9	Yu G, Luo H, Zhang N, Wang Y, Li Y, Huang H, Liu Y, Hu Y, Liu H, Zhang J, et al: Loss of p53 sensitizes cells to palmitic acid-induced apoptosis by reactive oxygen species accumulation. Int J Mol Sci. 20(6268)2019.PubMed/NCBI View Article : Google Scholar
10	Okamoto Y, Kihara S, Ouchi N, Nishida M, Arita Y, Kumada M, Ohashi K, Sakai N, Shimomura I, Kobayashi H, et al: Adiponectin reduces atherosclerosis in apolipoprotein E-deficient mice. Circulation. 106:2767–2770. 2002.PubMed/NCBI View Article : Google Scholar
11	Whitehead JP, Richards AA, Hickman IJ, Macdonald GA and Prins JB: Adiponectin - a key adipokine in the metabolic syndrome. Diabetes Obes Metab. 8:264–280. 2006.PubMed/NCBI View Article : Google Scholar
12	Aksin S and Andan C: Protein-9 (CTRP9) levels associated with C1q tumor necrosis factor in obese preeclamptic, non-obese preeclamptic, obese and normal pregnant women. J Matern Fetal Neonatal Med. 34:2540–2547. 2021.PubMed/NCBI View Article : Google Scholar
13	Zuo A, Zhao X, Li T, Li J, Lei S, Chen J, Xu D, Song C, Liu T, Li C and Guo Y: CTRP9 knockout exaggerates lipotoxicity in cardiac myocytes and high-fat diet-induced cardiac hypertrophy through inhibiting the LKB1/AMPK pathway. J Cell Mol Med. 24:2635–2647. 2020.PubMed/NCBI View Article : Google Scholar
14	Jia Y, Luo X, Ji Y, Xie J, Jiang H, Fu M and Li X: Circulating CTRP9 levels are increased in patients with newly diagnosed type 2 diabetes and correlated with insulin resistance. Diabetes Res Clin Pract. 131:116–123. 2017.PubMed/NCBI View Article : Google Scholar
15	Niemann B, Li L, Siegler D, Siegler BH, Knapp F, Hanna J, Aslam M, Kracht M, Schulz R and Rohrbach S: CTRP9 mediates protective effects in cardiomyocytes via AMPK- and adiponectin receptor-mediated induction of anti-oxidant response. Cells. 9(1229)2020.PubMed/NCBI View Article : Google Scholar
16	Zhao SJ, Shen YF, Li Q, He YJ, Zhang YK, Hu LP, Jiang YQ, Xu NW, Wang YJ, Li J, et al: SLIT2/ROBO1 axis contributes to the Warburg effect in osteosarcoma through activation of SRC/ERK/c-MYC/PFKFB2 pathway. Cell Death Dis. 9(390)2018.PubMed/NCBI View Article : Google Scholar
17	Myerburg MM, King JD Jr, Oyster NM, Fitch AC, Magill A, Baty CJ, Watkins SC, Kolls JK, Pilewski JM and Hallows KR: AMPK agonists ameliorate sodium and fluid transport and inflammation in cystic fibrosis airway epithelial cells. Am J Respir Cell Mol Biol. 42:676–684. 2010.PubMed/NCBI View Article : Google Scholar
18	Ren L, Sun D, Zhou X, Yang Y, Huang X and Li Y, Wang C and Li Y: Chronic treatment with the modified Longdan Xiegan Tang attenuates olanzapine-induced fatty liver in rats by regulating hepatic de novo lipogenesis and fatty acid beta-oxidation-associated gene expression mediated by SREBP-1c, PPAR-alpha and AMPK-alpha. J Ethnopharmacol. 232:176–187. 2019.PubMed/NCBI View Article : Google Scholar
19	Kohjima M, Higuchi N, Kato M, Kotoh K, Yoshimoto T, Fujino T, Yada M, Yada R, Harada N, Enjoji M, et al: SREBP-1c, regulated by the insulin and AMPK signaling pathways, plays a role in nonalcoholic fatty liver disease. Int J Mol Med. 21:507–511. 2008.PubMed/NCBI
20	Jayachandran M, Zhang T, Wu Z, Liu Y and Xu B: Isoquercetin regulates SREBP-1C via AMPK pathway in skeletal muscle to exert antihyperlipidemic and anti-inflammatory effects in STZ induced diabetic rats. Mol Biol Rep. 47:593–602. 2020.PubMed/NCBI View Article : Google Scholar
21	Lee G, Jang H, Kim YY, Choe SS, Kong J, Hwang I, Park J, Im SS and Kim JB: SREBP1c-PAX4 axis mediates pancreatic β-Cell compensatory responses upon metabolic stress. Diabetes. 68:81–94. 2019.PubMed/NCBI View Article : Google Scholar
22	Larkin JC, Sears SB and Sadovsky Y: The influence of ligand-activated LXR on primary human trophoblasts. Placenta. 35:919–924. 2014.PubMed/NCBI View Article : Google Scholar
23	Tian L, Wen A, Dong S and Yan P: Molecular characterization of microtubule affinity-regulating kinase4 from sus scrofa and promotion of lipogenesis in primary porcine placental trophoblasts. Int J Mol Sci. 20(1206)2019.PubMed/NCBI View Article : Google Scholar
24	Kanehisa M and Goto S: KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Res. 28:27–30. 2000.PubMed/NCBI View Article : Google Scholar
25	Kanehisa M: Toward understanding the origin and evolution of cellular organisms. Protein Sci. 28:1947–1951. 2019.PubMed/NCBI View Article : Google Scholar
26	Kanehisa M, Furumichi M, Sato Y, Ishiguro-Watanabe M and Tanabe M: KEGG: Integrating viruses and cellular organisms. Nucleic Acids Res. 49D:D545–D551. 2021.PubMed/NCBI View Article : Google Scholar
27	Graham CH, Hawley TS, Hawley RG, MacDougall JR, Kerbel RS, Khoo N and Lala PK: Establishment and characterization of first trimester human trophoblast cells with extended lifespan. Exp Cell Res. 206:204–211. 1993.PubMed/NCBI View Article : Google Scholar
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
29	Li J, Zhang P, Li T, Liu Y, Zhu Q, Chen T, Liu T, Huang C, Zhang J, Zhang Y and Guo Y: CTRP9 enhances carotid plaque stability by reducing pro-inflammatory cytokines in macrophages. Biochem Biophys Res Commun. 458:890–895. 2015.PubMed/NCBI View Article : Google Scholar
30	Sun H, Zhu X, Zhou Y, Cai W and Qiu L: C1q/TNF-related protein-9 ameliorates Ox-LDL-induced endothelial dysfunction via PGC-1α/AMPK-mediated antioxidant enzyme induction. Int J Mol Sci. 18(1097)2017.PubMed/NCBI View Article : Google Scholar
31	Moradi N, Fadaei R, Emamgholipour S, Kazemian E, Panahi G, Vahedi S, Saed L and Fallah S: Association of circulating CTRP9 with soluble adhesion molecules and inflammatory markers in patients with type 2 diabetes mellitus and coronary artery disease. PLoS One. 13(e0192159)2018.PubMed/NCBI View Article : Google Scholar
32	Zhang P, Huang C, Li J, Li T, Guo H, Liu T, Li N, Zhu Q and Guo Y: Globular CTRP9 inhibits oxLDL-induced inflammatory response in RAW 264.7 macrophages via AMPK activation. Mol Cell Biochem. 417:67–74. 2016.PubMed/NCBI View Article : Google Scholar
33	Challier JC, Basu S, Bintein T, Minium J, Hotmire K, Catalano PM and Hauguel-de Mouzon S: Obesity in pregnancy stimulates macrophage accumulation and inflammation in the placenta. Placenta. 29:274–281. 2008.PubMed/NCBI View Article : Google Scholar
34	El-Haggar SM and Mostafa TM: Adipokines and biochemical changes in Egyptian obese subjects: Possible variation with sex and degree of obesity. Endocrine. 48:878–885. 2015.PubMed/NCBI View Article : Google Scholar
35	Jones HN, Jansson T and Powell TL: IL-6 stimulates system A amino acid transporter activity in trophoblast cells through STAT3 and increased expression of SNAT2. Am J Physiol Cell Physiol. 297:C1228–C1235. 2009.PubMed/NCBI View Article : Google Scholar
36	Aye IL, Jansson T and Powell TL: Interleukin-1β inhibits insulin signaling and prevents insulin-stimulated system A amino acid transport in primary human trophoblasts. Mol Cell Endocrinol. 381:46–55. 2013.PubMed/NCBI View Article : Google Scholar
37	Knöfler M, Mösl B, Bauer S, Griesinger G and Husslein P: TNF-alpha/TNFRI in primary and immortalized first trimester cytotrophoblasts. Placenta. 21:525–535. 2000.PubMed/NCBI View Article : Google Scholar
38	Kambara T, Ohashi K, Shibata R, Ogura Y, Maruyama S, Enomoto T, Uemura Y, Shimizu Y, Yuasa D, Matsuo K, et al: CTRP9 protein protects against myocardial injury following ischemia-reperfusion through AMP-activated protein kinase (AMPK)-dependent mechanism. J Biol Chem. 287:18965–18973. 2012.PubMed/NCBI View Article : Google Scholar
39	Yamaguchi S, Shibata R, Ohashi K, Enomoto T, Ogawa H, Otaka N, Hiramatsu-Ito M, Masutomi T, Kawanishi H, Murohara T and Ouchi N: C1q/TNF-related protein 9 promotes revascularization in response to ischemia via an eNOS-dependent manner. Front Pharmacol. 11(1313)2020.PubMed/NCBI View Article : Google Scholar
40	Yecies JL, Zhang HH, Menon S, Liu S, Yecies D, Lipovsky AI, Gorgun C, Kwiatkowski DJ, Hotamisligil GS, Lee CH and Manning BD: Akt stimulates hepatic SREBP1c and lipogenesis through parallel mTORC1-dependent and independent pathways. Cell Metab. 14:21–32. 2011.PubMed/NCBI View Article : Google Scholar
41	Kwon D, Kim SH, Son SW, Seo J, Jeong TB, Kim KM, Jung JC, Jung MS, Lee YH and Jung YS: Germinated soybean embryo extract ameliorates fatty liver injury in high-fat diet-fed obese mice. Pharmaceuticals (Basel). 13(380)2020.PubMed/NCBI View Article : Google Scholar