Intervention by Picroside II on FFAs Induced Lipid Accumulation and Lipotoxicity in HepG2 Cells

Overview

Journal J Ayurveda Integr Med

Publisher Elsevier

Date 2021 Aug 6

PMID 34353693

Citations 1

Authors

Hiteshi Dhami-Shah

Rama Vaidya

Manasi Talwadekar

Eisha Shaw

Shobha Udipi

Ullas Kolthur-Seetharam

Ashok D B Vaidya

Affiliations

Soon will be listed here.

Abstract

Background: Accumulation of free fatty acids (FFAs) in hepatocytes is a hallmark of liver dysfunction and non-alcoholic fatty liver disease (NAFLD). Excessive deposition of FFAs alters lipid metabolism pathways increasing the oxidative stress and mitochondrial dysfunction. Attenuating hepatic lipid accumulation, oxidative stress, and improving mitochondrial function could provide potential targets in preventing progression of non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH). Earlier studies with Picrorhiza kurroa extract have shown reduction in hepatic damage and fatty acid infiltration in several experimental models and also clinically in viral hepatitis. Thus, the effect of P. kurroa's phytoactive, picroside II, needed mechanistic investigation in appropriate in vitro liver cell model.

Objective(s): To study the effect of picroside II on FFAs accumulation, oxidative stress and mitochondrial function with silibinin as a positive control in in vitro NAFLD model.

Materials And Methods: HepG2 cells were incubated with FFAs-1000μM in presence and absence of Picroside II-10 μM for 20 hours.

Results: HepG2 cells incubated with FFAs-1000μM lead to increased lipid accumulation. Picroside II-10μM attenuated FFAs-induced lipid accumulation (33%), loss of mitochondrial membrane potential (ΔΨm), ATP depletion, and production of reactive oxygen species (ROS). A concomitant increase in cytochrome C at transcription and protein levels was observed. An increase in expression of MnSOD, catalase, and higher levels of tGSH and GSH:GSSG ratios underlie the ROS salvaging activity of picroside II.

Conclusion: Picroside II significantly attenuated FFAs-induced-lipotoxicity. The reduction in ROS, increased antioxidant enzymes, and improvement in mitochondrial function underlie the mechanisms of action of picroside II. These findings suggest a need to develop an investigational drug profile of picroside II for NAFLD as a therapeutic strategy. This could be evaluated through the fast-track path of reverse pharmacology.

Citing Articles

Alterations in Glutathione Redox Homeostasis in Metabolic Dysfunction-Associated Fatty Liver Disease: A Systematic Review.

Cesarini L, Grignaffini F, Alisi A, Pastore A Antioxidants (Basel). 2025; 13(12.

PMID: 39765791 PMC: 11672975. DOI: 10.3390/antiox13121461.

Picrorhiza kurroa, Royle ex Benth:Traditional uses, phytopharmacology, and translational potential in therapy of fatty liver disease.

Raut A, Dhami-Shah H, Phadke A, Shindikar A, Udipi S, Joshi J J Ayurveda Integr Med. 2022; 14(1):100558.

PMID: 35659739 PMC: 10105242. DOI: 10.1016/j.jaim.2022.100558.

References

De Rautlin De La Roy Y, Messedi N, Grollier G, Grignon B . Kinetics of bactericidal activity of antibiotics measured by luciferin-luciferase assay. J Biolumin Chemilumin. 1991; 6(3):193-201. DOI: 10.1002/bio.1170060310. View

Cole B, Feaver R, Wamhoff B, Dash A . Non-alcoholic fatty liver disease (NAFLD) models in drug discovery. Expert Opin Drug Discov. 2017; 13(2):193-205. DOI: 10.1080/17460441.2018.1410135. View

Dhami-Shah H, Vaidya R, Udipi S, Raghavan S, Abhijit S, Mohan V . Picroside II attenuates fatty acid accumulation in HepG2 cells via modulation of fatty acid uptake and synthesis. Clin Mol Hepatol. 2017; 24(1):77-87. PMC: 5875197. DOI: 10.3350/cmh.2017.0039. View

Rahman I, Kode A, Biswas S . Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat Protoc. 2007; 1(6):3159-65. DOI: 10.1038/nprot.2006.378. View

Matteoni C, Younossi Z, Gramlich T, Boparai N, Liu Y, McCullough A . Nonalcoholic fatty liver disease: a spectrum of clinical and pathological severity. Gastroenterology. 1999; 116(6):1413-9. DOI: 10.1016/s0016-5085(99)70506-8. View

Garcia-Ruiz I, Fernandez-Moreira D, Solis-Munoz P, Rodriguez-Juan C, Diaz-Sanjuan T, Munoz-Yague T . Mitochondrial complex I subunits are decreased in murine nonalcoholic fatty liver disease: implication of peroxynitrite. J Proteome Res. 2010; 9(5):2450-9. DOI: 10.1021/pr9011427. View

Perez-Carreras M, del Hoyo P, Martin M, Rubio J, Martin A, Castellano G . Defective hepatic mitochondrial respiratory chain in patients with nonalcoholic steatohepatitis. Hepatology. 2003; 38(4):999-1007. DOI: 10.1053/jhep.2003.50398. View

Begriche K, Igoudjil A, Pessayre D, Fromenty B . Mitochondrial dysfunction in NASH: causes, consequences and possible means to prevent it. Mitochondrion. 2006; 6(1):1-28. DOI: 10.1016/j.mito.2005.10.004. View

Rector R, Thyfault J, Uptergrove G, Morris E, Naples S, Borengasser S . Mitochondrial dysfunction precedes insulin resistance and hepatic steatosis and contributes to the natural history of non-alcoholic fatty liver disease in an obese rodent model. J Hepatol. 2010; 52(5):727-36. PMC: 3070177. DOI: 10.1016/j.jhep.2009.11.030. View

10.

Cousin S, Hugl S, Wrede C, Kajio H, Myers Jr M, Rhodes C . Free fatty acid-induced inhibition of glucose and insulin-like growth factor I-induced deoxyribonucleic acid synthesis in the pancreatic beta-cell line INS-1. Endocrinology. 2001; 142(1):229-40. DOI: 10.1210/endo.142.1.7863. View

11.

Pessayre D, Fromenty B . NASH: a mitochondrial disease. J Hepatol. 2005; 42(6):928-40. DOI: 10.1016/j.jhep.2005.03.004. View

12.

Antarkar D, Tathed P, Vaidya A . A pilot phase II trial with arogya-wardhani and Punarnavadi-Kwath in viral hepatitis. Panminerva Med. 1978; 20(3):157-60. View

13.

Chavez-Tapia N, Rosso N, Tiribelli C . In vitro models for the study of non-alcoholic fatty liver disease. Curr Med Chem. 2011; 18(7):1079-84. DOI: 10.2174/092986711794940842. View

14.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

15.

Chalasani N, Deeg M, Crabb D . Systemic levels of lipid peroxidation and its metabolic and dietary correlates in patients with nonalcoholic steatohepatitis. Am J Gastroenterol. 2004; 99(8):1497-502. DOI: 10.1111/j.1572-0241.2004.30159.x. View

16.

Dowman J, Tomlinson J, Newsome P . Pathogenesis of non-alcoholic fatty liver disease. QJM. 2009; 103(2):71-83. PMC: 2810391. DOI: 10.1093/qjmed/hcp158. View

17.

Angulo P . Nonalcoholic fatty liver disease. N Engl J Med. 2002; 346(16):1221-31. DOI: 10.1056/NEJMra011775. View

18.

Visen P, Shukla B, Patnaik G, Dhawan B . Prevention of galactosamine-induced hepatic damage by picroliv: study on bile flow and isolated hepatocytes (ex vivo). Planta Med. 1993; 59(1):37-41. DOI: 10.1055/s-2006-959600. View

19.

Younossi Z, Anstee Q, Marietti M, Hardy T, Henry L, Eslam M . Global burden of NAFLD and NASH: trends, predictions, risk factors and prevention. Nat Rev Gastroenterol Hepatol. 2017; 15(1):11-20. DOI: 10.1038/nrgastro.2017.109. View

20.

Distelmaier F, Koopman W, Testa E, de Jong A, Swarts H, Mayatepek E . Life cell quantification of mitochondrial membrane potential at the single organelle level. Cytometry A. 2008; 73(2):129-38. DOI: 10.1002/cyto.a.20503. View