نوع مقاله : مقاله پژوهشی
نویسندگان
1 دانشجوی دکتری، گروه زیستشناسی، دانشگاه پیام نور، تهران، ایران
2 استادیار گروه بیوشیمی، دانشکده پزشکی، دانشگاه علوم پزشکی ایران، تهران، ایران
3 استادیار گروه زیستشناسی، دانشگاه پیام نور، تهران، ایران
4 دانشجوی دکتری، گروه بیوشیمی، دانشکده پزشکی، دانشگاه علوم پزشکی ایران، تهران، ایران
چکیده
زمینه و هدف: بیماری کبد چرب غیرالکلی (NAFLD) بهعنوان یک بیماری شایع مرتبط با اختلالات متابولیک، به دلیل افزایش تجمع اسیدهای چرب اشباع مانند پالمیتات، منجر به اختلال در متابولیسم لیپید میشود. این پژوهش به بررسی اثر پالمیتات بر تجمع قطرات چربی و ژنهای مرتبط با انتقال معکوس کلسترول در سلولهای کبدی HepG2 میپردازد.
مواد و روشها: سلولهای HepG2 در شرایط استاندارد کشت داده شدند. تجمع قطرات چربی با رنگآمیزی Oil Red O، سطح کلسترول و تریگلیسرید با کیتهای استاندارد و تغییرات بیان ژنهای (ATP-Binding Cassette Transporter A1 and G1) ABCG1 وABCA1، (Apolipoprotein A1)Apo-A1، (Liver X Receptor) LXR و (Scavenger Receptor Class B Type ) SR-B1 با روش RT-qPCR ارزیابی گردید.
یافتهها: تیمار سلولهای HepG2 با پالمیتات با افزایش تجمع قطرات چربی سیتوپلاسمی و درعینحال با کاهش قابلتوجهی در زندهمانی سلولها همراه بود (P<0.0001). سطح کلسترول و تریگلیسرید سلولها نیز به دنبال تیمار با پالمیتات نسبت به گروه کنترل افزایش یافت (P<0.05). همچنین، پالمیتات باعث کاهش بیان ABCA1 و SR-BI (P<0.05) و افزایش بیانABCG1 (P<0.05) شد، اما تأثیری بر بیان LXR و Apo-A1 نداشت (P>0.05).
نتیجهگیری: پالمیتات با کاهش بیان ژنهای ABCA1 و SR-B1، فرآیند انتقال معکوس کلسترول را مختل کرده و باعث تجمع لیپید و افزایش سطوح لیپید داخل سلولی میشود. افزایش بیان ABCG1 بهعنوان یک پاسخ جبرانی نتوانست این اختلالات را جبران کند. این یافتهها تأکید میکند که پالمیتات از طریق تأثیر بر مسیرهای متابولیکی و التهابی میتواند به اختلال در هموستاز لیپید و بیماریهای متابولیک مرتبط با آن منجر شود.
تازه های تحقیق
https://scholar.google.com/citations?user=J6F7R7kAAAAJ&hl=fa
https://pubmed.ncbi.nlm.nih.gov/?term=Elham+Jafarpour
https://scholar.google.com/citations?user=g4DxSh4AAAAJ&hl=en&oi=sra
https://scholar.google.com/citations?user=g4DxSh4AAAAJ&hl=en&oi=sra
https://scholar.google.com/scholar?hl=en&as_sdt=0,5&q=Habiballah+Nazem&btnG=
https://pubmed.ncbi.nlm.nih.gov/22801281/
https://scholar.google.com/citations?user=0GBL7C8AAAAJ&hl=en&oi=sra
https://pubmed.ncbi.nlm.nih.gov/?term=Zolfaghar+Lotfi
https://scholar.google.com/citations?user=5LjBil0AAAAJ&hl=en&oi=ao
https://pubmed.ncbi.nlm.nih.gov/?term=Yaser+Mohammadi
کلیدواژهها
موضوعات
عنوان مقاله [English]
The Effect Of Palmitate On The Expression Levels Of Genes Involved In Reverse Cholesterol Transport And Lipid Metabolism In HepG2 Cells
نویسندگان [English]
- Elham Jafarpour 1
- Elham Bahreini 2
- Habiballah Nazem 3
- Zolfaghar Lotfi 3
- Yaser Mohammadi 4
1 student, Department of Biology, Payame Noor University, Tehran, Iran
2 Assistant Professor, Department of Biochemistry, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran
3 Assistant Professor, Department of Biology, Payame Noor University, Tehran, Iran
4 PhD student, Department of Biochemistry, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran
چکیده [English]
Introduction: Non-alcoholic fatty liver disease (NAFLD) is associated with metabolic disorders that disrupt lipid and cholesterol metabolism due to the accumulation of saturated fatty acids, such as palmitate. This study investigates the effects of palmitate on lipid droplet accumulation and the expression of genes involved in reverse cholesterol transport (RCT) in HepG2 liver cells.
Materials and Methods: HepG2 cells were cultured under standard conditions. Lipid droplet accumulation was assessed using Oil Red O staining. Cellular cholesterol and triglyceride levels were measured using standard kits, and gene expression changes for ABCA1 and ABCG (ATP-Binding Cassette Transporter A1 and G1, respectively), APO-A1 (Apolipoprotein A1), SR-BI (Scavenger Receptor Class B Type I), and LXR (Liver X Receptor) were evaluated by RT-qPCR.
Results: Treatment of HepG2 cells with palmitate resulted in a significant increase in lipid droplet accumulation, which inversely correlated with cell viability (P<0.0001). Cholesterol and triglyceride levels also significantly increased following palmitate treatment compared to the control group (P<0.05). Additionally, palmitate decreased the expression of ABCA1 and SR-BI (P < 0.05) while increasing the expression of ABCG1 (P < 0.05). No significant changes were observed in the expression of APOA1 and LXR (P > 0.05).
Conclusion: Palmitate disrupts reverse cholesterol transport by downregulating ABCA1 and SR-BI expression, leading to lipid accumulation and elevated intracellular cholesterol and triglyceride levels. The upregulation of ABCG1 as a compensatory response does not adequately counter these disruptions. These findings suggest that palmitate, through its effects on metabolic and inflammatory pathways, contributes to disturbances in lipid homeostasis and associated metabolic disorders.
کلیدواژهها [English]
- Lipid Droplets؛ Non-alcoholic Fatty Liver Disease؛
- Palmitic Acid
- ؛ Reverse Cholesterol Transport
- Vancells Lujan P, Viñas Esmel E, Sacanella Meseguer E. Overview of Non-Alcoholic Fatty Liver Disease (NAFLD) and the Role of Sugary Food Consumption and Other Dietary Components in Its Development. Nutrients. 2021;13(5). doi: 10.3390/nu13051442.
- Henry L, Paik J, Younossi ZM. Review article: the epidemiologic burden of non-alcoholic fatty liver disease across the world. Aliment Pharmacol Ther. 2022;56(6):942-56. doi: 10.1111/apt.17158.
- Leamy AK, Egnatchik RA, Young JD. Molecular mechanisms and the role of saturated fatty acids in the progression of non-alcoholic fatty liver disease. Prog Lipid Res. 2013;52(1):165-74. doi: 10.1016/j.plipres.2012.10.004.
- Murru E, Manca C, Carta G, Banni S. Impact of Dietary Palmitic Acid on Lipid Metabolism. Front Nutr. 2022;9:861664. doi: 10.3389/fnut.2022.861664.
- van Rooijen MA, Plat J, Blom WAM, Zock PL, Mensink RP. Dietary stearic acid and palmitic acid do not differently affect ABCA1-mediated cholesterol efflux capacity in healthy men and postmenopausal women: A randomized controlled trial. Clin Nutr. 2021;40(3):804-11. doi: 10.1016/j.clnu.2020.08.016.
- Ouimet M, Barrett TJ, Fisher EA. HDL and Reverse Cholesterol Transport. Circ Res. 2019;124(10):1505-18. doi: 10.1161/CIRCRESAHA.119.312617.
- Yu XH, Tang CK. ABCA1, ABCG1, and Cholesterol Homeostasis. Adv Exp Med Biol. 2022;1377:95-107. doi: 10.1007/978-981-19-1592-5_7.
- Yvan-Charvet L, Wang N, Tall AR. Role of HDL, ABCA1, and ABCG1 transporters in cholesterol efflux and immune responses. Arterioscler Thromb Vasc Biol. 2010;30(2):139-43. doi: 10.1161/ATVBAHA.108.179283
- Bilotta MT, Petillo S, Santoni A, Cippitelli M. Liver X Receptors: Regulators of Cholesterol Metabolism, Inflammation, Autoimmunity, and Cancer. Front Immunol. 2020;11:584303. doi: 10.3389/fimmu.2020.584303.
- Zhao NQ, Li XY, Wang L, Feng ZL, Li XF, Wen YF, et al. Palmitate induces fat accumulation by activating C/EBPβ-mediated G0S2 expression in HepG2 cells. World J Gastroenterol. 2017;23(43):7705-15. doi: 10.3748/wjg.v23.i43.7705.
- Teixeira FS, Pimentel LL, Vidigal S, Azevedo-Silva J, Pintado ME, Rodríguez-Alcalá LM. Differential Lipid Accumulation on HepG2 Cells Triggered by Palmitic and Linoleic Fatty Acids Exposure. Molecules. 2023;28(5). doi: 10.3390/molecules28052367.
- Lyu J, Imachi H, Fukunaga K, Sato S, Kobayashi T, Dong T, et al. Role of ATP-binding cassette transporter A1 in suppressing lipid accumulation by glucagon-like peptide-1 agonist in hepatocytes. Mol Metab. 2020;34:16-26. doi: 10.1016/j.molmet.2019.12.015.
- Jeon BH, Lee YH, Yun MR, Kim SH, Lee BW, Kang ES, et al. Increased expression of ATP-binding cassette transporter A1 (ABCA1) as a possible mechanism for the protective effect of cilostazol against hepatic steatosis. Metabolism. 2015;64(11):1444-53. doi: 10.1016/j.metabol.2015.07.014.
- Rein-Fischboeck L, Krautbauer S, Eisinger K, Pohl R, Meier EM, Weiss TS, et al. Hepatic scavenger receptor BI is associated with type 2 diabetes but unrelated to human and murine non-alcoholic fatty liver disease. Biochem Biophys Res Commun. 2015;467(2):377-82. doi: 10.1016/j.bbrc.2015.09.149.
- Spady DK, Kearney DM, Hobbs HH. Polyunsaturated fatty acids up-regulate hepatic scavenger receptor B1 (SR-BI) expression and HDL cholesteryl ester uptake in the hamster. J Lipid Res. 1999;40(8):1384-94. doi: 10.1016/S0022-2275(20)33380-0
- Rocca C, De Bartolo A, Guzzi R, Crocco MC, Rago V, Romeo N, et al. Palmitate-Induced Cardiac Lipotoxicity Is Relieved by the Redox-Active Motif of SELENOT through Improving Mitochondrial Function and Regulating Metabolic State. Cells. 2023;12(7). doi: 10.3390/cells12071042.
- Liu C, Zhou B, Meng M, Zhao W, Wang D, Yuan Y, et al. FOXA3 induction under endoplasmic reticulum stress contributes to non-alcoholic fatty liver disease. J Hepatol. 2021;75(1):150-62. doi: 10.1016/j.jhep.2021.01.042.
- Li Y, Xu Y, Jadhav K, Zhu Y, Yin L, Zhang Y. Hepatic Forkhead Box Protein A3 Regulates ApoA-I (Apolipoprotein A-I) Expression, Cholesterol Efflux, and Atherogenesis. Arterioscler Thromb Vasc Biol. 2019;39(8):1574-87. doi: 10.1161/ATVBAHA.119.312610.
- Vargas-Alarcón G, Fragoso JM, Ramírez-Bello J, Posadas-Sánchez R. FOXA3 Polymorphisms Are Associated with Metabolic Parameters in Individuals with Subclinical Atherosclerosis and Healthy Controls-The GEA Mexican Study. Biomolecules. 2022;12(5). doi: 10.3390/biom12050601.
- Shen WJ, Azhar S, Kraemer FB. SR-B1: A Unique Multifunctional Receptor for Cholesterol Influx and Efflux. Annu Rev Physiol. 2018;80:95-116. doi: 10.1146/annurev-physiol-021317-121550.
- Rivera K, Quiñones V, Amigo L, Santander N, Salas-Pérez F, Xavier A, et al. Lipoprotein receptor SR-B1 deficiency enhances adipose tissue inflammation and reduces susceptibility to hepatic steatosis during diet-induced obesity in mice. Biochim Biophys Acta Mol Cell Biol Lipids. 2021;1866(6):158909. doi: 10.1016/j.bbalip.2021.158909.
- Moore JM, Bell EL, Hughes RO, Garfield AS. ABC transporters: human disease and pharmacotherapeutic potential. Trends Mol Med. 2023;29(2):152-72. doi: 10.1016/j.molmed.2022.11.001.
- Kennedy MA, Barrera GC, Nakamura K, Baldán A, Tarr P, Fishbein MC, et al. ABCG1 has a critical role in mediating cholesterol efflux to HDL and preventing cellular lipid accumulation. Cell Metab. 2005;1(2):121-31. doi: 10.1016/j.cmet.2005.01.002.
- Wang X, Collins HL, Ranalletta M, Fuki IV, Billheimer JT, Rothblat GH, et al. Macrophage ABCA1 and ABCG1, but not SR-BI, promote macrophage reverse cholesterol transport in vivo. J Clin Invest. 2007;117(8):2216-24. doi: 10.1172/JCI32057.
- Korbecki J, Bajdak-Rusinek K. The effect of palmitic acid on inflammatory response in macrophages: an overview of molecular mechanisms. Inflamm Res. 2019;68(11):915-32. doi: 10.1007/s00011-019-01273-5.
- Ahn SB, Jang K, Jun DW, Lee BH, Shin KJ. Expression of liver X receptor correlates with intrahepatic inflammation and fibrosis in patients with nonalcoholic fatty liver disease. Dig Dis Sci. 2014;59(12):2975-82. doi: 10.1007/s10620-014-3289-x.
- Parks E, Yki-Järvinen H, Hawkins M. Out of the frying pan: dietary saturated fat influences nonalcoholic fatty liver disease. J Clin Invest. 2017;127(2):454-6. doi: 10.1007/s11154-019-09512-0.
- Madaudo C, Bono G, Ortello A, Astuti G, Mingoia G, Galassi AR, et al. Dysfunctional High-Density Lipoprotein Cholesterol and Coronary Artery Disease: A Narrative Review. J Pers Med. 2024;14(9). doi: 10.3390/jpm14090996.
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