Purinergic 2X7 receptor is involved in adipogenesis and lipid degradation

Li,Jing; Gong,Linxia; Xu,Qiaolan

doi:10.3892/etm.2021.11004

Purinergic 2X7 receptor is involved in adipogenesis and lipid degradation

Authors:
- Jing Li
- Linxia Gong
- Qiaolan Xu
View Affiliations

Affiliations: Pediatric Department, Yancheng Third People's Hospital, Yancheng School of Clinical Medicine of Nanjing Medical University, Yancheng, Jiangsu 224000, P.R. China, Pediatric Department, Nanjing Integrated Traditional Chinese and Western Medicine Hospital, Nanjing, Jiangsu 210024, P.R. China
Published online on: November 25, 2021 https://doi.org/10.3892/etm.2021.11004
Article Number: 81
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Obesity and dyslipidemia are two metabolic syndrome disorders that have serious effects on the health of patients. Purinergic 2X receptor ligand‑gated ion channel 7 (P2X7R) has been reported to play a role in regulating lipid storage and metabolism. However, the role and potential mechanism of P2X7R in adipogenesis and lipid degradation remain unknown. In the present study, a mouse model of obesity was established by feeding mice a high‑fat diet, and the 3T3‑L1 cell line was used to analyze the function of P2X7R in vitro. Reverse transcription‑quantitative PCR and western blot analyses were performed to detect the expression levels of P2X7R, sterol regulatory element‑binding protein 1 (SREBP1) and other associated transcription factors. Bioinformatics analysis was used to predict the potential target gene of P2X7R and a dual luciferase reporter assay was used to confirm this prediction. Oil Red O staining was used to evaluate the adipogenic capacity of preadipocytes. AdipoRed assay, cholesterol assay and a free glycerol reagent were used to measure the expression levels of triglyceride (TGs), total cholesterol (TC) and glycerin, respectively. The results indicated that P2X7R was highly expressed in obese mice and that it was involved in adipogenic differentiation in vitro. SREBP1 enhanced the transcription activities of P2X7R to promote its expression. Inhibition of P2X7R significantly reduced the adipogenic capacity of preadipocytes, decreased the expression levels of adipogenesis‑associated transcription factors (peroxisome proliferator‑activated receptor γ, CCAAT‑enhancer‑binding protein α and fatty‑acid‑binding protein 4), enhanced the expression levels of lipolytic enzymes (adipose triglyceride lipase, phosphorylated hormone‑sensitive lipase and monoacylglycerol lipase) and regulated the expression of TG, TC and glycerin in mature 3T3‑L1 cells. These effects were reversed by a small interfering RNA targeting Wnt3a. Therefore, the results suggested that P2X7R, the transcription activities of which were regulated by SREBP1, regulated adipogenesis and lipid degradation by targeting SREBP1, indicating its potential effects on obesity‑associated metabolism.

View Figures

View References

1	Bou M, Todorcevic M, Rodriguez J, Capilla E, Gutierrez J and Navarro I: Interplay of adiponectin, TNFα and insulin on gene expression, glucose uptake and PPARү, AKT and TOR pathways in rainbow trout cultured adipocytes. Gen Comp Endocrinol. 205:218–225. 2014.PubMed/NCBI View Article : Google Scholar
2	Saxton SN, Clark BJ, Withers SB, Eringa EC and Heagerty AM: Mechanistic links between obesity, diabetes, and blood pressure: Role of perivascular adipose tissue. Physiol Rev. 99:1701–1763. 2019.PubMed/NCBI View Article : Google Scholar
3	Kahn CR, Wang G and Lee KY: Altered adipose tissue and adipocyte function in the pathogenesis of metabolic syndrome. J Clin Invest. 129:3990–4000. 2019.PubMed/NCBI View Article : Google Scholar
4	Bleich SN, Vercammen KA, Zatz LY, Frelier JM, Ebbeling CB and Peeters A: Interventions to prevent global childhood overweight and obesity: A systematic review. Lancet Diabetes Endocrinol. 6:332–346. 2018.PubMed/NCBI View Article : Google Scholar
5	Bhupathiraju SN and Hu FB: Epidemiology of obesity and diabetes and their cardiovascular complications. Circ Res. 118:1723–1735. 2016.PubMed/NCBI View Article : Google Scholar
6	Gallagher EJ and LeRoith D: Obesity and diabetes: The increased risk of cancer and cancer-related mortality. Physiol Rev. 95:727–748. 2015.PubMed/NCBI View Article : Google Scholar
7	Tang QQ and Lane MD: Adipogenesis: From stem cell to adipocyte. Annu Rev Biochem. 81:715–736. 2012.PubMed/NCBI View Article : Google Scholar
8	Haczeyni F, Bell-Anderson KS and Farrell GC: Causes and mechanisms of adipocyte enlargement and adipose expansion. Obes Rev. 19:406–420. 2018.PubMed/NCBI View Article : Google Scholar
9	Sawamoto A, Nakanishi M, Okuyama S, Furukawa Y and Nakajima M: Heptamethoxyflavone inhibits adipogenesis via enhancing PKA signaling. Eur J Pharmacol. 865(172758)2019.PubMed/NCBI View Article : Google Scholar
10	Im DU, Kim SC, Chau GC and Um SH: Carbamazepine enhances adipogenesis by inhibiting Wnt/β-catenin expression. Cells. 8(1460)2019.PubMed/NCBI View Article : Google Scholar
11	Prestwich TC and Macdougald OA: Wnt/beta-catenin signaling in adipogenesis and metabolism. Curr Opin Cell Biol. 19:612–617. 2007.PubMed/NCBI View Article : Google Scholar
12	Chen X, Ayala I, Shannon C, Fourcaudot M, Acharya NK, Jenkinson CP, Heikkinen S and Norton L: The diabetes gene and Wnt pathway effector TCF7L2 regulates adipocyte development and function. Diabetes. 67:554–568. 2018.PubMed/NCBI View Article : Google Scholar
13	Guo Y, Mishra A, Weng T, Chintagari NR, Wang Y, Zhao C, Huang C and Liu L: Wnt3a mitigates acute lung injury by reducing P2X7 receptor-mediated alveolar epithelial type I cell death. Cell Death Dis. 5(e1286)2014.PubMed/NCBI View Article : Google Scholar
14	Sindhavajiva PR, Sastravaha P, Arksornnukit M and Pavasant P: Purinergic 2X7 receptor activation regulates WNT signaling in human mandibular-derived osteoblasts. Arch Oral Biol. 81:167–174. 2017.PubMed/NCBI View Article : Google Scholar
15	Gnoni A, Siculella L, Paglialonga G, Damiano F and Giudetti AM: 3,5-diiodo-L-thyronine increases de novo lipogenesis in liver from hypothyroid rats by SREBP-1 and ChREBP-mediated transcriptional mechanisms. IUBMB Life. 71:863–872. 2019.PubMed/NCBI View Article : Google Scholar
16	Vadrot N, Duband-Goulet I, Cabet E, Attanda W, Barateau A, Vicart P, Gerbal F, Briand N, Vigouroux C, Oldenburg AR, et al: The p. R482W substitution in A-type lamins deregulates SREBP1 activity in Dunnigan-type familial partial lipodystrophy. Hum Mol Genet. 24:2096–2109. 2015.PubMed/NCBI View Article : Google Scholar
17	Hou XX, Dong HR, Sun LJ, Yang M, Cheng H and Chen YP: Purinergic 2X7 receptor is involved in the podocyte damage of obesity-related glomerulopathy via activating nucleotide-binding and oligomerization domain-like receptor protein 3 inflammasome. Chin Med J (Engl). 131:2713–2725. 2018.PubMed/NCBI View Article : Google Scholar
18	North RA: Molecular physiology of P2X receptors. Physiol Rev. 82:1013–1067. 2002.PubMed/NCBI View Article : Google Scholar
19	Su QQ, Tian YY, Liu ZN, Ci LL and Lv XW: Purinergic P2X7 receptor blockade mitigates alcohol-induced steatohepatitis and intestinal injury by regulating MEK1/2-ERK1/2 signaling and egr-1 activity. Int Immunopharmacol. 66:52–61. 2019.PubMed/NCBI View Article : Google Scholar
20	Beaucage KL, Xiao A, Pollmann SI, Grol MW, Beach RJ, Holdsworth DW, Sims SM, Darling MR and Dixon SJ: Loss of P2X7 nucleotide receptor function leads to abnormal fat distribution in mice. Purinergic Signal. 10:291–304. 2014.PubMed/NCBI View Article : Google Scholar
21	Gao H, Li D, Yang P, Zhao L, Wei L, Chen Y and Ruan XZ: Suppression of CD36 attenuates adipogenesis with a reduction of P2X7 expression in 3T3-L1 cells. Biochem Biophys Res Commun. 491:204–208. 2017.PubMed/NCBI View Article : Google Scholar
22	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
23	Han Y, Lee SH, Lee IS and Lee KY: Regulatory effects of 4-methoxychalcone on adipocyte differentiation through PPARγ activation and reverse effect on TNF-α in 3T3-L1 cells. Food Chem Toxicol. 106:17–24. 2017.PubMed/NCBI View Article : Google Scholar
24	Lee CW, Seo JY, Lee J, Choi JW, Cho S, Bae JY, Sohng JK, Kim SO, Kim J and Park YI: 3-O-Glucosylation of quercetin enhances inhibitory effects on the adipocyte differentiation and lipogenesis. Biomed Pharmacother. 95:589–598. 2017.PubMed/NCBI View Article : Google Scholar
25	Ma J, Liao H, Lin Y, Huang K, Zhu J and Wang Y: Molecular characterization, expression analysis of Chemerin gene and its potential role in intramuscular adipocyte differentiation of goat. Anim Biotechnol. 31:382–390. 2020.PubMed/NCBI View Article : Google Scholar
26	Rosen ED and MacDougald OA: Adipocyte differentiation from the inside out. Nat Rev Mol Cell Biol. 7:885–896. 2006.PubMed/NCBI View Article : Google Scholar
27	Moseti D, Regassa A and Kim WK: Molecular regulation of adipogenesis and potential anti-adipogenic bioactive molecules. Int J Mol Sci. 17(124)2016.PubMed/NCBI View Article : Google Scholar
28	Zhang L, Zhang L, Wang X and Si H: Anti-adipogenic effects and mechanisms of ginsenoside Rg3 in pre-adipocytes and obese mice. Front Pharmacol. 8(113)2017.PubMed/NCBI View Article : Google Scholar
29	Jin H, Lee K, Chei S, Oh HJ, Lee KP and Lee BY: Ecklonia stolonifera extract suppresses lipid accumulation by promoting lipolysis and adipose browning in high-fat diet-induced obese male mice. Cells. 9(871)2020.PubMed/NCBI View Article : Google Scholar
30	Di Virgilio F, Dal Ben D, Sarti AC, Giuliani AL and Falzoni S: The P2X7 receptor in infection and inflammation. Immunity. 47:15–31. 2017.PubMed/NCBI View Article : Google Scholar
31	Coccurello R and Volonte C: P2X7 Receptor in the management of energy homeostasis: Implications for obesity, dyslipidemia, and insulin resistance. Front Endocrinol (Lausanne). 11(199)2020.PubMed/NCBI View Article : Google Scholar
32	Calderon-Dominguez M, Mir JF, Fucho R, Weber M, Serra D and Herrero L: Fatty acid metabolism and the basis of brown adipose tissue function. Adipocyte. 5:98–118. 2015.PubMed/NCBI View Article : Google Scholar
33	Moon MH, Jeong JK, Lee YJ, Seol JW, Ahn DC, Kim IS and Park SY: 18β-Glycyrrhetinic acid inhibits adipogenic differentiation and stimulates lipolysis. Biochem Biophys Res Commun. 420:805–810. 2012.PubMed/NCBI View Article : Google Scholar
34	Seo YJ, Jin H, Lee K, Song JH, Chei S, Oh HJ, Oh JH and Lee BY: Cardamonin suppresses lipogenesis by activating protein kinase A-mediated browning of 3T3-L1 cells. Phytomedicine. 65(153064)2019.PubMed/NCBI View Article : Google Scholar
35	Lee JE, Schmidt H, Lai B and Ge K: Transcriptional and epigenomic regulation of adipogenesis. Mol Cell Biol. 39:e00601–e00618. 2019.PubMed/NCBI View Article : Google Scholar
36	Walther TC and Farese RV Jr: Lipid droplets and cellular lipid metabolism. Annu Rev Biochem. 81:687–714. 2012.PubMed/NCBI View Article : Google Scholar
37	Suh HJ, Cho SY, Kim EY and Choi HS: Blockade of lipid accumulation by silibinin in adipocytes and zebrafish. Chem Biol Interact. 227:53–62. 2015.PubMed/NCBI View Article : Google Scholar
38	Clevers H: Wnt/beta-catenin signaling in development and disease. Cell. 127:469–480. 2006.PubMed/NCBI View Article : Google Scholar
39	Xi FX, Wei CS, Xu YT, Ma L, He YL, Shi XE, Yang GS and Yu TY: MicroRNA-214-3p targeting Ctnnb1 promotes 3T3-L1 preadipocyte differentiation by interfering with the Wnt/β-catenin signaling pathway. Int J Mol Sci. 20(1816)2019.PubMed/NCBI View Article : Google Scholar
40	Kang S, Bennett CN, Gerin I, Rapp LA, Hankenson KD and Macdougald OA: Wnt signaling stimulates osteoblastogenesis of mesenchymal precursors by suppressing CCAAT/enhancer-binding protein alpha and peroxisome proliferator-activated receptor gamma. J Biol Chem. 282:14515–14524. 2007.PubMed/NCBI View Article : Google Scholar
41	Liu J and Farmer SR: Regulating the balance between peroxisome proliferator-activated receptor gamma and beta-catenin signaling during adipogenesis. A glycogen synthase kinase 3beta phosphorylation-defective mutant of beta-catenin inhibits expression of a subset of adipogenic genes. J Biol Chem. 279:45020–45027. 2004.PubMed/NCBI View Article : Google Scholar
42	Liu J, Wang H, Zuo Y and Farmer SR: Functional interaction between peroxisome proliferator-activated receptor gamma and beta-catenin. Mol Cell Biol. 26:5827–5837. 2006.PubMed/NCBI View Article : Google Scholar