1α,25(OH)₂D₃ Regulates the TGF-β1/Samd Signaling Pathway Inhibition of Hepatic Stellate Cell Activation

Abstract

Objective

To investigate the effect of 1α,25(OH)₂D₃ on hepatic stellate cells and the mechanism of the TGF-β1/Smad signaling pathway.

Methods

LX2 cells were treated with TGF-β1 and different concentrations of 1α,25(OH)₂D₃. Cell proliferation was assessed using the CCK8 assay to determine the optimal concentration of 1α,25(OH)₂D₃ activity. The cell cycle and apoptotic rates were evaluated using flow cytometry. The expressions of Samd2, Samd3, Samd4, and Samd7 was assessed by western blotting, whereas the expression of MMP1, MMP13, and TIMP-1 was detected by qPCR.

Results

Compared with the control group, the 1α,25(OH)₂D₃ group had a higher apoptotic rate of LX2 cells, the cell cycle was blocked from the G1 stage to the S stage, the expressions of Samd2, Samd7, MMP1, and MMP13 increased, while the expressions of Samd3, Samd4, and TIMP-1 decreased.

Conclusion

1α,25(OH)₂D₃ inhibits hepatic stellate cell activation and exerts anti-hepatic fibrosis effects by downregulating the expression of Samd3, Samd4, TIMP-1 and upregulating the expression of Samd2, Samd4, MMP1, and MMP13.

Keywords

hepatic stellate cell - 1α,25(OH)₂D₃ - TGF-β1 - Samds protein

Publication History

Received: 29 January 2024

Accepted: 05 November 2024

Article published online:
15 January 2025

• References

• 1 Zhao M, Wang L, Wang M. et al. Targeting fibrosis, mechanisms and cilinical trials. Signal Transduct Target Ther 2022; 7: 206
• 2 Wong MCS, Huang JLW, George J. et al. The changing epidemiology of liver diseases in the Asia-Pacific region. Nat Rev Gastroenterol Hepatol 2019; 16: 57-73
• 3 Wong RJ, Aguilar M, Cheung R. et al. Nonalcoholic steatohepatitis is the second leading etiology of liver disease among adults awaiting liver transplantation in the United States. Gastroenterology 2015; 148: 547-555
• 4 Pimpin L, Cortez-Pinto H, Negro F. et al. EASL HEPAHEALTH Steering Committee. Burden of liver disease in Europe: epidemiology and analysis of risk factors to identify prevention policies. J Hepatol 2018; 69: 718-735
• 5 Roehlen N, Crouchet E, Baumert TF. Liver fibrosis: mechanistic concepts and therapeutic perspectives [J]. Cells 2020; 9: 875
• 6 Akcora BÖ, Dathathri E, Ortiz-Perez A. et al. TG101348, a selective JAK2 antagonist, ameliorates hepatic fibrogenesis in vivo [J]. FASEB J 2019; 33: 9466-9475
• 7 Xu S, Mao Y, Wu J. et al. TGF-β/Smad and JAK/STAT pathways are involved in the anti-fibrotic effects of propylene glycol alginate sodium sulphate on hepatic fibrosis [J]. J Cell Mol Med 2020; 24: 5224-5237
• 8 Yuan X, Gong Z, Wang B. et al. Astragaloside Inhibits Hepatic Fibrosis by Modulation of TGF-β1/Smad Signaling Pathway. Evid Based Complement Alternat Med 2018; 2018: 3231647
• 9 Elashiry M, Elsayed R, Elashiry MM. et al. Proteomic Characterization, Biodistribution, and Functional Studies of Immune-Therapeutic Exosomes: Implications for Inflammatory Lung Diseases. Front Immunol 2021; 12: 636222
• 10 Barchetta I, Cimini FA, Cavallo MG. Vitamin D and metabolic dysfunction-associated fatty liver disease (MAFLD): an update [J]. Nutrients 2020; 12: 3302
• 11 Jahn D, Dorbath D, Kircher S. et al. Beneficial Effects of Vitamin D Treatment in an Obese Mouse Model of Non-Alcoholic Steatohepatitis. Nutrients [J] 2019; 11: 77
• 12 Abramovitch S, Sharvit E, Weisman Y. et al. Vitamin D inhibits development of liver fibrosis in an animal model but cannot ameliorate established cirrhosis [J]. Am J Physiol Gastrointest Liver Physiol 2015; 308: G112-G120
• 13 Pop TL, Sîrbe C, Benţa G. et al. The Role of Vitamin D and Vitamin D Binding Protein in Chronic Liver Diseases.The Role of Vitamin D and Vitamin D Binding Protein in Chronic Liver Diseases [J]. Int J Mol Sci 2022; 23: 10705
• 14 Lu W, Li X, Liu N. et al. Vitamin D alleviates liver fibrosis by inhibiting histidine-rich calcium binding protein (HRC) [J]. Chem Biol Interact 2021; 334: 109355
• 15 Branton MH, Kopp JB. “TGF-beta and fibrosis.”. Microbes and infection vol [J] 1999; 1: 1349-1365
• 16 Wang W, Zhang Y, Jiang Y. et al. Exploration of potential mechanism of Rougan formula against hepatic fibrosis by network analysis and experimental assessment [J]. J Ethnopharmacol 2023; 304: 115960
• 17 Chen Y, Liao W, Zhu Z. et al. Essential oil from the raw and vinegar-processed Rhizoma Curcumae ameliorate CCL₄-induced liver fibrosis: integrating network pharmacology and molecular mechanism evaluation [J]. Food Funct 2021; 12: 4199-4220
• 18 Corachán A, Trejo MG, Carbajo-García MC. et al. Vitamin D as an effective treatment in human uterine leiomyomas independent of mediator complex subunit 12 mutation [J]. Fertil Steril 2021; 115: 512-521
• 19 Hijazi N, Shi Z, Rockey DC. Paxillin regulates liver fibrosis via actin polymerization and ERK activation in hepatic stellate cells [J]. J Cell Sci 2023; 136: jcs261122
• 20 Zhu L, Kong M, Han YP. et al. Spontaneous liver fibrosis induced by long term dietary vitamin D deficiency in adult mice is related to chronic inflammation and enhanced apoptosis [J]. Canadian journal of physiology and pharmacology 2015; 93: 385-394
• 21 Yoshida K, Murata M, Yamaguchi T. et al. TGF-β/Smad signaling during hepatic fibro-carcinogenesis (review). International journal of oncology [J] 2014; 45: 1363-1371
• 22 Abramovitch S, Dahan-Bachar L, Sharvit E. et al. Vitamin D inhibits proliferation and profibrotic marker expression in hepatic stellate cells and decreases thioacetamide-induced liver fibrosis in rats [J]. Gut 2011; 60: 1728-1737
• 23 Duarte S, Baber J, Fujii T. et al. Matrix metalloproteinases in liver injury, repair and fibrosis. Matrix biology [J]. Journal of the International Society for Matrix Biology 2015; 44–46: 147-156
• 24 Komolmit P, Kimtrakool S, Suksawatamnuay S. et al. Vitamin D supplementation improves serum markers associated with hepatic fibrogenesis in chronic hepatitis C patients: A randomized, double-blind, placebo-controlled study [J]. Scientific reports 2017; 7: 8905