Top

Published in:

Open Access 01-08-2020 | Original Contribution

In vitro effects of sitosterol and sitostanol on mitochondrial respiration in human brown adipocytes, myotubes and hepatocytes

Published in: European Journal of Nutrition | Issue 5/2020

Abstract

Purpose

Lowering of LDL cholesterol levels by plant sterols and stanols is associated with decreased risk of cardiovascular disease in humans. Plant sterols and stanols also lower triacylglycerol (TG). However, it is not fully understood how reduction in TG is achieved and what the full potential of plant sterols and stanols is on whole-body metabolism. We here hypothesize that high levels of plant sterols and stanols stimulate whole-body energy expenditure, which can be attributed to changes in mitochondrial function of brown adipose tissue (BAT), skeletal muscle and liver.

Methods

Phytosterolemic mice were fed chow diets for 32 weeks to examine whole-body weight gain. In vitro, 24-h incubation were performed in adipocytes derived from human BAT, human myotubes or HepG2 human hepatocytes using sitosterol or sitostanol. Following mitochondrial function was assessed using seahorse bioanalyzer.

Results

Chow feeding in phytosterolemic mice resulted in diminished increase in body weight compared to control mice. In vitro, sitosterol or sitostanol did not change mitochondrial function in adipocytes derived from human BAT or in cultured human myotubes. Interestingly, maximal mitochondrial function in HepG2 human hepatocytes was decreased following sitosterol or sitostanol incubation, however, only when mitochondrial function was assessed in low glucose-containing medium.

Conclusions

Beneficial in vivo effects of plant sterols and stanols on lipid and lipoprotein metabolism are well recognized. Our results indicate that alterations in human mitochondrial function are apparently not involved to explain these beneficial effects.

Gylling H, Plat J, Turley S, Ginsberg HN, Ellegard L, Jessup W, Jones PJ, Lutjohann D, Maerz W, Masana L, Silbernagel G, Staels B, Boren J, Catapano AL, De Backer G, Deanfield J, Descamps OS, Kovanen PT, Riccardi G, Tokgozoglu L, Chapman MJ (2014) Plant sterols and plant stanols in the management of dyslipidaemia and prevention of cardiovascular disease. Atherosclerosis 232(2):346–360. https://doi.org/10.1016/j.atherosclerosis.2013.11.043 CrossRefPubMed

Plat J, Brufau G, Dallinga-Thie GM, Dasselaar M, Mensink RP (2009) A plant stanol yogurt drink alone or combined with a low-dose statin lowers serum triacylglycerol and non-HDL cholesterol in metabolic syndrome patients. J Nutr 139(6):1143–1149. https://doi.org/10.3945/jn.108.103481 CrossRefPubMed

Hegele RA, Ginsberg HN, Chapman MJ, Nordestgaard BG, Kuivenhoven JA, Averna M, Boren J, Bruckert E, Catapano AL, Descamps OS, Hovingh GK, Humphries SE, Kovanen PT, Masana L, Pajukanta P, Parhofer KG, Raal FJ, Ray KK, Santos RD, Stalenhoef AF, Stroes E, Taskinen MR, Tybjaerg-Hansen A, Watts GF, Wiklund O (2014) The polygenic nature of hypertriglyceridaemia: implications for definition, diagnosis, and management. Lancet Diabetes Endocrinol 2(8):655–666. https://doi.org/10.1016/S2213-8587(13)70191-8 CrossRefPubMed

Bansal S, Buring JE, Rifai N, Mora S, Sacks FM, Ridker PM (2007) Fasting compared with nonfasting triglycerides and risk of cardiovascular events in women. JAMA 298(3):309–316. https://doi.org/10.1001/jama.298.3.309 CrossRefPubMed

Nordestgaard BG, Benn M, Schnohr P, Tybjaerg-Hansen A (2007) Nonfasting triglycerides and risk of myocardial infarction, ischemic heart disease, and death in men and women. JAMA 298(3):299–308. https://doi.org/10.1001/jama.298.3.299 CrossRefPubMed

Demonty I, Ras RT, van der Knaap HC, Meijer L, Zock PL, Geleijnse JM, Trautwein EA (2013) The effect of plant sterols on serum triglyceride concentrations is dependent on baseline concentrations: a pooled analysis of 12 randomised controlled trials. Eur J Nutr 52(1):153–160. https://doi.org/10.1007/s00394-011-0297-x CrossRefPubMed

Theuwissen E, Plat J, van der Kallen CJ, van Greevenbroek MM, Mensink RP (2009) Plant stanol supplementation decreases serum triacylglycerols in subjects with overt hypertriglyceridemia. Lipids 44(12):1131–1140. https://doi.org/10.1007/s11745-009-3367-6 CrossRefPubMed

Schonewille M, Brufau G, Shiri-Sverdlov R, Groen AK, Plat J (2014) Serum TG-lowering properties of plant sterols and stanols are associated with decreased hepatic VLDL secretion. J Lipid Res 55(12):2554–2561. https://doi.org/10.1194/jlr.M052407 CrossRefPubMedPubMedCentral

Plat J, Mensink RP (2009) Plant stanol esters lower serum triacylglycerol concentrations via a reduced hepatic VLDL-1 production. Lipids 44(12):1149–1153. https://doi.org/10.1007/s11745-009-3361-z CrossRefPubMedPubMedCentral

10.

Baumgartner S, Mensink RP, Plat J (2016) Effects of a plant sterol or stanol enriched mixed meal on postprandial lipid metabolism in healthy subjects. PLoS ONE 11(9):e0160396. https://doi.org/10.1371/journal.pone.0160396 CrossRefPubMedPubMedCentral

11.

Cannon B, Nedergaard J (2004) Brown adipose tissue: function and physiological significance. Physiol Rev 84(1):277–359. https://doi.org/10.1152/physrev.00015.2003 CrossRefPubMed

12.

Cypess AM, Kahn CR (2010) Brown fat as a therapy for obesity and diabetes. Curr Opin Endocrinol Diabetes Obes 17(2):143–149. https://doi.org/10.1097/MED.0b013e328337a81f CrossRefPubMedPubMedCentral

13.

Schrauwen P, van Marken Lichtenbelt WD (2016) Combatting type 2 diabetes by turning up the heat. Diabetologia 59(11):2269–2279. https://doi.org/10.1007/s00125-016-4068-3 CrossRefPubMedPubMedCentral

14.

Ouellet V, Labbe SM, Blondin DP, Phoenix S, Guerin B, Haman F, Turcotte EE, Richard D, Carpentier AC (2012) Brown adipose tissue oxidative metabolism contributes to energy expenditure during acute cold exposure in humans. J Clin Investig 122(2):545–552. https://doi.org/10.1172/JCI60433 CrossRefPubMed

15.

Bartelt A, Bruns OT, Reimer R, Hohenberg H, Ittrich H, Peldschus K, Kaul MG, Tromsdorf UI, Weller H, Waurisch C, Eychmuller A, Gordts PL, Rinninger F, Bruegelmann K, Freund B, Nielsen P, Merkel M, Heeren J (2011) Brown adipose tissue activity controls triglyceride clearance. Nat Med 17(2):200–205. https://doi.org/10.1038/nm.2297 CrossRefPubMed

16.

Geerling JJ, Boon MR, van der Zon GC, van den Berg SA, van den Hoek AM, Lombes M, Princen HM, Havekes LM, Rensen PC, Guigas B (2014) Metformin lowers plasma triglycerides by promoting VLDL-triglyceride clearance by brown adipose tissue in mice. Diabetes 63(3):880–891. https://doi.org/10.2337/db13-0194 CrossRefPubMed

17.

Mackay DS, Jones PJ, Myrie SB, Plat J, Lutjohann D (2014) Methodological considerations for the harmonization of non-cholesterol sterol bio-analysis. J Chromatogr B Anal Technol Biomed Life Sci 957:116–122. https://doi.org/10.1016/j.jchromb.2014.02.052 CrossRef

18.

van der Krieken SE, Popeijus HE, Mensink RP, Plat J (2017) Link between ER-stress, PPAR-alpha activation, and BET inhibition in relation to apolipoprotein A–I transcription in HepG2 cells. J Cell Biochem 118(8):2161–2167. https://doi.org/10.1002/jcb.25858 CrossRefPubMed

19.

Sparks LM, Bosma M, Brouwers B, van de Weijer T, Bilet L, Schaart G, Moonen-Kornips E, Eichmann TO, Lass A, Hesselink MKC, Schrauwen P (2014) Reduced incorporation of fatty acids into triacylglycerol in myotubes from obese individuals with type 2 diabetes. Diabetes 63(5):1583–1593. https://doi.org/10.2337/db13-1123 CrossRefPubMedPubMedCentral

20.

Broeders EP, Nascimento EB, Havekes B, Brans B, Roumans KH, Tailleux A, Schaart G, Kouach M, Charton J, Deprez B, Bouvy ND, Mottaghy F, Staels B, van Marken Lichtenbelt WD, Schrauwen P (2015) The bile acid chenodeoxycholic acid increases human brown adipose tissue activity. Cell Metab 22(3):418–426. https://doi.org/10.1016/j.cmet.2015.07.002 CrossRefPubMed

21.

Klett EL, Lu K, Kosters A, Vink E, Lee MH, Altenburg M, Shefer S, Batta AK, Yu H, Chen J, Klein R, Looije N, Oude-Elferink R, Groen AK, Maeda N, Salen G, Patel SB (2004) A mouse model of sitosterolemia: absence of Abcg8/sterolin-2 results in failure to secrete biliary cholesterol. BMC Med 2:5. https://doi.org/10.1186/1741-7015-2-5 CrossRefPubMedPubMedCentral

22.

DeFronzo RA (1988) Lilly lecture 1987. The triumvirate: beta-cell, muscle, liver. A collusion responsible for NIDDM. Diabetes 37(6):667–687CrossRef

23.

Nascimento EBM, Sparks LM, Divoux A, van Gisbergen MW, Broeders EPM, Jorgensen JA, Schaart G, Bouvy ND, van Marken Lichtenbelt WD, Schrauwen P (2018) Genetic markers of brown adipose tissue identity and in vitro brown adipose tissue activity in humans. Obesity 26(1):135–140. https://doi.org/10.1002/oby.22062 CrossRefPubMed

24.

Panov A, Kubalik N, Brooks BR, Shaw CA (2010) In vitro effects of cholesterol beta-d-glucoside, cholesterol and cycad phytosterol glucosides on respiration and reactive oxygen species generation in brain mitochondria. J Membr Biol 237(2–3):71–77. https://doi.org/10.1007/s00232-010-9307-9 CrossRefPubMed

25.

Shi C, Wu F, Xu J (2013) Incorporation of beta-sitosterol into mitochondrial membrane enhances mitochondrial function by promoting inner mitochondrial membrane fluidity. J Bioenerg Biomembr 45(3):301–305. https://doi.org/10.1007/s10863-012-9495-3 CrossRefPubMed

26.

Wong HS, Chen N, Leong PK, Ko KM (2014) beta-Sitosterol enhances cellular glutathione redox cycling by reactive oxygen species generated from mitochondrial respiration: protection against oxidant injury in H9c2 cells and rat hearts. Phytother Res 28(7):999–1006. https://doi.org/10.1002/ptr.5087 CrossRefPubMed

27.

Kim YS, Li XF, Kang KH, Ryu B, Kim SK (2014) Stigmasterol isolated from marine microalgae Navicula incerta induces apoptosis in human hepatoma HepG2 cells. BMB Rep 47(8):433–438CrossRef

28.

Roussi S, Gosse F, Aoude-Werner D, Zhang X, Marchioni E, Geoffroy P, Miesch M, Raul F (2007) Mitochondrial perturbation, oxidative stress and lysosomal destabilization are involved in 7beta-hydroxysitosterol and 7beta-hydroxycholesterol triggered apoptosis in human colon cancer cells. Apoptosis 12(1):87–96. https://doi.org/10.1007/s10495-006-0485-y CrossRefPubMed

29.

De Smet E, Mensink RP, Plat J (2012) Effects of plant sterols and stanols on intestinal cholesterol metabolism: suggested mechanisms from past to present. Mol Nutr Food Res 56(7):1058–1072. https://doi.org/10.1002/mnfr.201100722 CrossRefPubMed

30.

Plat J, Hendrikx T, Bieghs V, Jeurissen ML, Walenbergh SM, van Gorp PJ, De Smet E, Konings M, Vreugdenhil AC, Guichot YD, Rensen SS, Buurman WA, Greve JW, Lutjohann D, Mensink RP, Shiri-Sverdlov R (2014) Protective role of plant sterol and stanol esters in liver inflammation: insights from mice and humans. PLoS O ne 9(10):e110758. https://doi.org/10.1371/journal.pone.0110758 CrossRef

31.

Clayton PT, Bowron A, Mills KA, Massoud A, Casteels M, Milla PJ (1993) Phytosterolemia in children with parenteral nutrition-associated cholestatic liver disease. Gastroenterology 105(6):1806–1813CrossRef

32.

Clayton PT, Whitfield P, Iyer K (1998) The role of phytosterols in the pathogenesis of liver complications of pediatric parenteral nutrition. Nutrition 14(1):158–164CrossRef

33.

Gura KM, Duggan CP, Collier SB, Jennings RW, Folkman J, Bistrian BR, Puder M (2006) Reversal of parenteral nutrition-associated liver disease in two infants with short bowel syndrome using parenteral fish oil: implications for future management. Pediatrics 118(1):e197–e201. https://doi.org/10.1542/peds.2005-2662 CrossRefPubMed

34.

Gura KM, Lee S, Valim C, Zhou J, Kim S, Modi BP, Arsenault DA, Strijbosch RA, Lopes S, Duggan C, Puder M (2008) Safety and efficacy of a fish-oil-based fat emulsion in the treatment of parenteral nutrition-associated liver disease. Pediatrics 121(3):e678–e686. https://doi.org/10.1542/peds.2007-2248 CrossRefPubMed

Title: In vitro effects of sitosterol and sitostanol on mitochondrial respiration in human brown adipocytes, myotubes and hepatocytes
Publication date: 01-08-2020
Published in: European Journal of Nutrition / Issue 5/2020
Print ISSN: 1436-6207
Electronic ISSN: 1436-6215
DOI: https://doi.org/10.1007/s00394-019-02052-y

ACC 2024 Congress Coverage

Heart Failure Video

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

In vitro effects of sitosterol and sitostanol on mitochondrial respiration in human brown adipocytes, myotubes and hepatocytes

Abstract

Purpose

Methods

Results

Conclusions

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 5/2020

Long-term Paleolithic diet is associated with lower resistant starch intake, different gut microbiota composition and increased serum TMAO concentrations

Long-term dietary adherence and changes in dietary intake in coronary patients after intervention with a Mediterranean diet or a low-fat diet: the CORDIOPREV randomized trial

Maternal probiotic milk intake during pregnancy and breastfeeding complications in the Norwegian Mother and Child Cohort Study

Association between carbohydrate intake and fatty acids in the de novo lipogenic pathway in serum phospholipids and adipose tissue in a population of Swedish men

Associations among dietary seaweed intake, c-MYC rs6983267 polymorphism, and risk of colorectal cancer in a Korean population: a case–control study

Cross-sectional association between non-soy legume consumption, serum uric acid and hyperuricemia: the PREDIMED-Plus study

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page