Genistein alleviates chronic vascular inflammatory response via the miR‑21/NF‑κB p65 axis in lipopolysaccharide‑treated mice

Xie,Xiaolin; Cong,Li; Liu,Sujuan; Xiang,Liping; Fu,Xiaohua

doi:10.3892/mmr.2021.11831

Genistein alleviates chronic vascular inflammatory response via the miR‑21/NF‑κB p65 axis in lipopolysaccharide‑treated mice

Authors:
- Xiaolin Xie
- Li Cong
- Sujuan Liu
- Liping Xiang
- Xiaohua Fu
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Affiliations: Key Laboratory of Study and Discovery of Small Targeted Molecules of Hunan Province, Hunan Normal University, Changsha, Hunan 410013, P.R. China, Department of Basic Medicine, School of Medicine, Hunan Normal University, Changsha, Hunan 410013, P.R. China, Department of Pathology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China
Published online on: January 7, 2021 https://doi.org/10.3892/mmr.2021.11831
Article Number: 192
Copyright: © Xie et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Chronic vascular inflammatory response is an important pathological basis of cardiovascular disease. Genistein (GEN), a natural compound, exhibits anti‑inflammatory effects. The aim of the present study was to investigate the effects of GEN on lipopolysaccharide (LPS)‑induced chronic vascular inflammatory response in mice and explore the underlying anti‑inflammatory mechanisms. C57BL/6 mice were fed with a high‑fat diet combined with intraperitoneal injection of LPS to induce chronic vascular inflammation. The expression levels of TNF‑α, IL‑6 and microRNA (miR)‑21 in the vasculature were detected via reverse transcription‑quantitative (RT‑q)PCR. The protein levels of inducible nitric oxide synthase (iNOS) and NF‑κB p65 were detected via western blotting. NF‑κB p65 was also analyzed via immunohistochemistry and immunofluorescence (IF). In addition, after transfection with miR‑21 mimic or inhibitor for 24 h, vascular endothelial cells (VECs) were treated with GEN and LPS. RT‑qPCR and western blot analyses were performed to detect the expression of TNF‑α, IL‑6, miR‑21 and iNOS, and the protein levels of iNOS and NF‑κB p65, respectively. IF was used to measure NF‑κB p65 nuclear translocation. The results revealed that GEN significantly decreased the expression of inflammation‑associated vascular factors in LPS‑treated C57BL/6 mice, including TNF‑α, IL‑6, iNOS, NF‑κB p65 and miR‑21. Furthermore, miR‑21 antagomir enhanced the anti‑inflammatory effects of GEN. In LPS‑induced VECs, miR‑21 mimic increased inflammation‑associated factor expression and attenuated the anti‑inflammatory effects of GEN, whereas miR‑21 inhibitor induced opposing effects. Therefore, the results of the present study suggested that GEN inhibited chronic vascular inflammatory response in mice, which may be associated with the inhibition of VEC inflammatory injury via the miR‑21/NF‑κB p65 pathway.

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1	Libby P and Hansson GK: Inflammation and immunity in diseases of the arterial tree: Players and layers. Circ Res. 116:307–311. 2015. View Article : Google Scholar : PubMed/NCBI
2	Gimbrone MA Jr and García-Cardeña G: Endothelial cell dysfunction and the pathobiology of atherosclerosis. Circ Res. 118:620–636. 2016. View Article : Google Scholar : PubMed/NCBI
3	Ross R: Atherosclerosis-an inflammatory disease. N Engl J Med. 340:115–126. 1999. View Article : Google Scholar : PubMed/NCBI
4	Bacchi S, Palumbo P, Sponta A and Coppolino MF: Clinical pharmacology of non-steroidal anti-inflammatory drugs: A review. Antiinflamm Antiallergy Agents Med Chem. 11:52–64. 2012. View Article : Google Scholar : PubMed/NCBI
5	LiverTox: Clinical and research information on drug-induced liver injury (Internet). Bethesda (MD) National Institute of Diabetes and Digestive and Kidney Diseases. 316431762012.
6	Harirforoosh S, Asghar W and Jamali F: Adverse effects of nonsteroidal antiinflammatory drugs: An update of gastrointestinal, cardiovascular and renal complications. J Pharm Pharm Sci. 16:821–847. 2013. View Article : Google Scholar : PubMed/NCBI
7	Sutrisno S, Aprina H, Simanungkalit HM, Andriyani A, Barlianto W, Sujuti H, Santoso S, Dwijayasa PM, Wahyuni ES and Mustofa E: Genistein modulates the estrogen receptor and suppresses angiogenesis and inflammation in the murine model of peritoneal endometriosis. J Tradit Complement Med. 8:278–281. 2017. View Article : Google Scholar : PubMed/NCBI
8	Arakawa S, Inoue M, Kinouchi R, Morizumi S, Yamaguchi M, Shimazu Y and Takeda M: Dietary constituent genistein inhibits the hyperexcitability of trigeminal nociceptive neurons associated with mechanical hyperalgesia following orofacial inflammation. J Oral Biosci. 61:215–220. 2019. View Article : Google Scholar : PubMed/NCBI
9	Jaiswal N, Akhtar J, Singh SP, Badruddeen and Ahsan F: An overview on genistein and its various formulations. Drug Res (Stuttg). 69:305–313. 2019. View Article : Google Scholar : PubMed/NCBI
10	Liu B, Xu L, Yu X, Jiao X, Yan J, Li W and Guo M: Genistein inhibited estradiol-induced vascular endothelial cell injury by downregulating the FAK/Focal adhesion pathway. Cell Physiol Biochem. 49:2277–2292. 2018. View Article : Google Scholar : PubMed/NCBI
11	Han S, Wu H, Li W and Gao P: Protective effects of genistein in homocysteine-induced endothelial cell inflammatory injury. Mol Cell Biochem. 403:43–49. 2015. View Article : Google Scholar : PubMed/NCBI
12	Deretic V and Levine B: Autophagy balances inflammation in innate immunity. Autophagy. 14:243–251. 2018. View Article : Google Scholar : PubMed/NCBI
13	Deretic V and Klionsky DJ: Autophagy and inflammation: A special review issue. Autophagy. 14:179–180. 2018. View Article : Google Scholar : PubMed/NCBI
14	Zhang H, Yang X, Pang X, Zhao Z, Yu H and Zhou H: Genistein protects against ox-LDL-induced senescence through enhancing SIRT1/LKB1/AMPK-mediated autophagy flux in HUVECs. Mol Cell Biochem. 455:127–134. 2019. View Article : Google Scholar : PubMed/NCBI
15	Zhang H, Zhao Z, Pang X, Yang J, Yu H, Zhang Y, Zhou H and Zhao J: miR-34a/sirtuin-1/foxo3a is involved in genistein protecting against ox-LDL-induced oxidative damage in HUVECs. Toxicol Lett. 277:115–122. 2017. View Article : Google Scholar : PubMed/NCBI
16	Yi L, Chang M, Zhao Q, Zhou Z, Huang X, Guo F and Huan J: Genistein-3′-sodium sulphonate protects against lipopolysaccharide-induced lung vascular endothelial cell apoptosis and acute lung injury via BCL-2 signalling. J Cell Mol Med. 24:1022–1035. 2020. View Article : Google Scholar : PubMed/NCBI
17	Hajibabaie F, Kouhpayeh S, Mirian M, Rahimmanesh I, Boshtam M, Sadeghian L, Gheibi A, Khanahmad H and Shariati L: MicroRNAs as the actors in the atherosclerosis scenario. J Physiol Biochem. 76:1–12. 2020. View Article : Google Scholar : PubMed/NCBI
18	Li J, Li K and Chen X: Inflammation-regulatory microRNAs: Valuable targets for intracranial atherosclerosis. J Neurosci Res. 97:1242–1252. 2019. View Article : Google Scholar : PubMed/NCBI
19	Tao L, Xu X, Fang Y, Wang A, Zhou F, Shen Y and Li J: miR-21 targets jnk and ccr7 to modulate the inflammatory response of grass carp following bacterial infection. Fish Shellfish Immunol. 94:258–263. 2019. View Article : Google Scholar : PubMed/NCBI
20	Shoeibi S: Diagnostic and theranostic microRNAs in the pathogenesis of atherosclerosis. Acta Physiol (Oxf). 228:e133532020. View Article : Google Scholar : PubMed/NCBI
21	Sessa R, Seano G, di Blasio L, Gagliardi PA, Isella C, Medico E, Cotelli F, Bussolino F and Primo L: The miR-126 regulates angiopoietin-1 signaling and vessel maturation by targeting p85β. Biochim Biophys Acta. 1823:1925–1935. 2012. View Article : Google Scholar : PubMed/NCBI
22	Muramatsu F, Kidoya H, Naito H, Sakimoto S and Takakura N: MicroRNA-125b inhibits tube formation of blood vessels through translational suppression of VE-cadherin. Oncogene. 32:414–421. 2013. View Article : Google Scholar : PubMed/NCBI
23	Zhang Y, Liu D, Chen X, Li J, Li L, Bian Z, Sun F, Lu J, Yin Y, Cai X, et al: Secreted monocytic miR-150 enhances targeted endothelial cell migration. Mol Cell. 39:133–144. 2010. View Article : Google Scholar : PubMed/NCBI
24	Pordzik J, Pisarz K, De Rosa S, Jones AD, Eyileten C, Indolfi C, Malek L and Postula M: The potential role of platelet-related microRNAs in the development of cardiovascular events in high-risk populations, including diabetic patients: A review. Front Endocrinol (Lausanne). 9:742018. View Article : Google Scholar : PubMed/NCBI
25	Canfran-Duque A, Rotllan N, Zhang X, Fernandez-Fuertes M, Ramirez-Hidalgo C, Araldi E, Daimiel L, Busto R, Fernandez-Hernando C and Suárez Y: Macrophage deficiency of miR-21 promotes apoptosis, plaque necrosis, and vascular inflammation during atherogenesis. EMBO Mol Med. 9:1244–1262. 2017. View Article : Google Scholar : PubMed/NCBI
26	Chipont A, Esposito B, Challier I, Montabord M, Tedgui A, Mallat Z, Loyer X and Potteaux S: MicroRNA-21 deficiency alters the survival of Ly-6C^lo monocytes in ApoE^−/− mice and reduces early-stage atherosclerosis-brief report. Arterioscler Thromb Vasc Biol. 39:170–177. 2019. View Article : Google Scholar : PubMed/NCBI
27	Huang X, Yue Z, Wu J, Chen J, Wwang S, Wu J, Ren L, Zhang A, Deng P, Wang K, et al: MicroRNA-21 knockout exacerbates angiotensin II-induced thoracic aortic aneurysm and dissection in mice with abnormal transforming growth factor-β-SMAD3 signaling. Arterioscler Thromb Vasc Biol. 38:1086–1101. 2018. View Article : Google Scholar : PubMed/NCBI
28	Li K, Cui MZ, Zhang KW, Wang GQ and Zhai ST: Effect of miR-21 on rat thoracic aortic aneurysm model by regulating the expressions of MMP-2 and MMP-9. Eur Rev Med Pharmacol Sci. 24:878–884. 2020.PubMed/NCBI
29	Cong L, Yang S, Zhang Y, Cao J and Fu X: DFMG attenuates the activation of macrophages induced by coculture with LPC-injured HUVE12 cells via the TLR4/MyD88/NFKB signaling pathway. Int J Mol Med. 41:2619–2628. 2018.PubMed/NCBI
30	Cong L, Zhang Y, Huang H, Cao J and Fu X: DFMG reverses proliferation and migration of vascular smooth muscle cells induced by co-culture with injured vascular endothelial cells via suppression of the TLR4-mediated signaling pathway. Mol Med Rep. 17:5692–5699. 2018.PubMed/NCBI
31	Xiang X, Li L, Bo P, Kuang T, Liu S, Xie X, Guo S, Fu X and Zhang Y: 7-Difluoromethyl-5,4′dimethoxygenistein exerts antiangiogenic effects on acute promyelocytic leukemia HL-60 cells by inhibiting the TLR4/NF-κB signaling pathway. Mol Med Rep. 21:2251–2259. 2020.PubMed/NCBI
32	Lee WR, Kim KH, An HJ, Park YY, Kim KS, Lee CK, Min BK and Park KK: Effects of chimeric decoy oligodeoxynucleotide in the regulation of transcription factors NF-κB and Sp1 in an animal model of atherosclerosis. Basic Clin Pharmacol Toxicol. 112:236–243. 2013. View Article : Google Scholar : PubMed/NCBI
33	Kim SJ, Park JH, Kim KH, Lee WR, Kim KS and Park KK: Melittin inhibits atherosclerosis in LPS/high-fat treated mice through atheroprotective actions. J Atheroscler Thromb. 18:1117–1126. 2011. View Article : Google Scholar : PubMed/NCBI
34	Kim SJ, Park JH, Kim KH, Lee WR, Lee S, Kwon OC, Kim KS and Park KK: Effect of NF-κB decoy oligodeoxynucleotide on LPS/high-fat diet-induced atherosclerosis in an animal model. Basic Clin Pharmacol Toxicol. 107:925–930. 2010. View Article : Google Scholar : PubMed/NCBI
35	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. View Article : Google Scholar : PubMed/NCBI
36	Liu J, Wang X, Zheng M and Luan Q: Lipopolysaccharide from Porphyromonas gingivalis promotes autophagy of human gingival fibroblasts through the PI3K/Akt/mTOR signaling pathway. Life Sci. 211:133–139. 2018. View Article : Google Scholar : PubMed/NCBI
37	Khedoe PPSJ, de Kleijn S, van Oeveren-Rietdijk AM, Plomp JJ, de Boer HC, van Pel M, Rensen PCN, Berbee JFP and Hiemstra PS: Acute and chronic effects of treatment with mesenchymal stromal cells on LPS-induced pulmonary inflammation, emphysema and atherosclerosis development. PLoS One. 12:e01837412017. View Article : Google Scholar : PubMed/NCBI
38	Sheane BJ, Smyth P, Scott K, Aziz R, Buckley M, Lodge E, Kiely N, Kingston M, McGovern E, Healy M, et al: An association between microRNA-21 expression and vitamin D deficiency in coronary artery disease. Microrna. 4:57–63. 2015. View Article : Google Scholar : PubMed/NCBI
39	Nejad C, Pepin G, Behlke MA and Gantier MP: Modified polyadenylation-based RT-qPCR increases selectivity of amplification of 3′-MicroRNA isoforms. Front Genet. 9:112018. View Article : Google Scholar : PubMed/NCBI
40	Krützfeldt J, Rajewsky N, Braich R, Rajeev KG, Tuschl T, Manoharan M and Stoffel M: Silencing of microRNAs in vivo with ‘antagomirs’. Nature. 438:685–689. 2005. View Article : Google Scholar : PubMed/NCBI
41	Hermkens DMA, Stam OCG, de Wit NM, Fontijn RD, Jongejan A, Moerland PD, Mackaaij C, Waas ISE, Daemen MJAP and de Vries HE: Profiling the unique protective properties of intracranial arterial endothelial cells. Acta Neuropathol Commun. 7:1512019. View Article : Google Scholar : PubMed/NCBI
42	Zeng Y, Xu J, Hua YQ, Peng Y and Xu XL: MDM2 contributes to oxidized low-density lipoprotein-induced inflammation through modulation of mitochondrial damage in endothelial cells. Atherosclerosis. 305:1–9. 2020. View Article : Google Scholar : PubMed/NCBI
43	Virani SS, Alonso A, Benjamin EJ, Bittencourt MS, Callaway CW, Carson AP, Chamberlain AM, Chang AR, Cheng S, Delling FN, et al: Heart disease and stroke statistics-2020 update: A report from the American heart association. Circulation. 141:e139–e596. 2020. View Article : Google Scholar : PubMed/NCBI
44	Varela ML, Mogildea M, Moreno I and Lopes A: Acute inflammation and metabolism. Inflammation. 41:1115–1127. 2018. View Article : Google Scholar : PubMed/NCBI
45	Aghasafari P, George U and Pidaparti R: A review of inflammatory mechanism in airway diseases. Inflamm Res. 68:59–74. 2019. View Article : Google Scholar : PubMed/NCBI
46	Skaper SD, Facci L, Zusso M and Giusti P: An inflammation-centric view of neurological disease: Beyond the neuron. Front Cell Neurosci. 12:722018. View Article : Google Scholar : PubMed/NCBI
47	Tan H, Zhao J, Zhang H, Zhai Q and Chen W: Novel strains of Bacteroides fragilis and Bacteroides ovatus alleviate the LPS-induced inflammation in mice. Appl Microbiol Biotechnol. 103:2353–2365. 2019. View Article : Google Scholar : PubMed/NCBI
48	Wu X, Zhang LL, Yin LB, Fu YJ, Jiang YJ, Ding HB, Chu ZX, Shang H and Zhang ZN: Deregulated microRNA-21 expression in monocytes from HIV-infected patients contributes to elevated IP-10 secretion in HIV infection. Front Immunol. 8:11222017. View Article : Google Scholar : PubMed/NCBI
49	Lu Y, An Y, Lv C, Ma W, Xi Y and Xiao R: Dietary soybean isoflavones in Alzheimer's disease prevention. Asia Pac J Clin Nutr. 27:946–954. 2018.PubMed/NCBI
50	Gholampour F, Mohammadi Z, Karimi Z and Owji SM: Protective effect of genistein in a rat model of ischemic acute kidney injury. Gene. 753:1447892020. View Article : Google Scholar : PubMed/NCBI
51	Das G, Paramithiotis S, Sundaram Sivamaruthi B, Wijaya CH, Suharta S, Sanlier N, Shin HS and Patra JK: Traditional fermented foods with anti-aging effect: A concentric review. Food Res Int. 134:1092692020. View Article : Google Scholar : PubMed/NCBI
52	Petry FDS, Coelho BP, Gaelzer MM, Kreutz F, Guma FTCR, Salbego CG and Trindade VMT: Genistein protects against amyloid-beta-induced toxicity in SH-SY5Y cells by regulation of Akt and Tau phosphorylation. Phytother Res. 34:796–807. 2020. View Article : Google Scholar : PubMed/NCBI
53	Ahmad Z, Ng CT, Fong LY, Bakar NA, Hussain NH, Ang KP, Ee GC and Hakim MN: Cryptotanshinone inhibits TNF-α-induced early atherogenic events in vitro. J Physiol Sci. 66:213–220. 2016. View Article : Google Scholar : PubMed/NCBI
54	Unver N and McAllister F: IL-6 family cytokines: Key inflammatory mediators as biomarkers and potential therapeutic targets. Cytokine Growth Factor Rev. 41:10–17. 2018. View Article : Google Scholar : PubMed/NCBI
55	Richards J, El-Hamamsy I, Chen S, Sarang Z, Sarathchandra P, Yacoub MH, Chester AH and Butcher JT: Side-specific endothelial-dependent regulation of aortic valve calcification: Interplay of hemodynamics and nitric oxide signaling. Am J Pathol. 182:1922–1931. 2013. View Article : Google Scholar : PubMed/NCBI
56	Cassini-Vieira P, Araujo FA, da Costa Dias FL, Russo RC, Andrade SP, Teixeira MM and Barcelos LS: iNOS activity modulates inflammation, angiogenesis, and tissue fibrosis in polyether-polyurethane synthetic implants. Mediators Inflamm. 2015:1384612015. View Article : Google Scholar : PubMed/NCBI
57	Bellucci A, Bubacco L, Longhena F, Parrella E, Faustini G, Porrini V, Bono F, Missale C and Pizzi M: Nuclear Factor-κB Dysregulation and α-synuclein pathology: Critical interplay in the pathogenesis of Parkinson's disease. Front Aging Neurosci. 12:682020. View Article : Google Scholar : PubMed/NCBI
58	Zhang Q, Lenardo MJ and Baltimore D: 30 Years of NF-κB: A blossoming of relevance to human pathobiology. Cell. 168:37–57. 2017. View Article : Google Scholar : PubMed/NCBI
59	Skvortsova K, Iovino N and Bogdanovic O: Functions and mechanisms of epigenetic inheritance in animals. Nat Rev Mol Cell Biol. 19:774–790. 2018. View Article : Google Scholar : PubMed/NCBI
60	Hulshoff MS, Del Monte-Nieto G, Kovacic J and Krenning G: Non-coding RNA in endothelial-to-mesenchymal transition. Cardiovasc Res. 115:1716–1731. 2019. View Article : Google Scholar : PubMed/NCBI
61	Singh RP, Massachi I, Manickavel S, Singh S, Rao NP, Hasan S, Mc Curdy DK, Sharma S, Wong D, Hahn BH and Rehimi H: The role of miRNA in inflammation and autoimmunity. Autoimmun Rev. 12:1160–1165. 2013. View Article : Google Scholar : PubMed/NCBI
62	Das A, Ganesh K, Khanna S, Sen CK and Roy S: Engulfment of apoptotic cells by macrophages: A role of microRNA-21 in the resolution of wound inflammation. J Immunol. 192:1120–1129. 2014. View Article : Google Scholar : PubMed/NCBI
63	Xue Z, Xi Q, Liu H, Guo X, Zhang J, Zhang Z, Li Y, Yang G, Zhou D, Yang H, et al: miR-21 promotes NLRP3 inflammasome activation to mediate pyroptosis and endotoxic shock. Cell Death Dis. 10:4612019. View Article : Google Scholar : PubMed/NCBI