Impact of lithocholic acid on the osteogenic and adipogenic differentiation balance of bone marrow mesenchymal stem cells_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

WANG Cui ¹ , LI Jiao ² , LU Lingyun ³ , LIU Lu ¹ ,  YU Xijie ¹

1. Laboratory of Endocrinology & Metabolism/Department of Endocrinology & Metabolism, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P. R. China;
2. Department of Infectious Diseases, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P. R. China;
3. Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P. R. China;

Corresponding author：

YU Xijie, Email: xijieyu@hotmail.com

Keywords：

Lithocholic acid; bone marrow mesenchymal stem cells; osteogenic differentiation; adipogenic differentiation; osteoporosis; mouse

DOI：

10.7507/1002-1892.202308050

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To Investigate the effects of lithocholic acid (LCA) on the balance between osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods Twelve 10-week-old SPF C57BL/6J female mice were randomly divided into an experimental group (undergoing bilateral ovariectomy) and a control group (only removing the same volume of adipose tissue around the ovaries), with 6 mice in each group. The body mass was measured every week after operation. After 4 weeks post-surgery, the weight of mouse uterus was measured, femur specimens of the mice were taken for micro-CT scanning and three-dimensional reconstruction to analyze changes in bone mass. Tibia specimens were taken for HE staining to calculate the number and area of bone marrow adipocytes in the marrow cavity area. ELISA was used to detect the expression of bone turnover markers in the serum. Liver samples were subjected to real-time fluorescence quantitative PCR (RT-qPCR) to detect the expression of key genes related to bile acid metabolism, including cyp7a1, cyp7b1, cyp8b1, and cyp27a1. BMSCs were isolated by centrifugation from 2 C57BL/6J female mice (10-week-old). The third-generation cells were exposed to 0, 1, 10, and 100 μmol/L LCA, following which cell viability was evaluated using the cell counting kit 8 assay. Subsequently, alkaline phosphatase (ALP) staining and oil red O staining were conducted after 7 days of osteogenic and adipogenic induction. RT-qPCR was employed to analyze the expressions of osteogenic-related genes, namely ALP, Runt-related transcription factor 2 (Runx2), and osteocalcin (OCN), as well as adipogenic-related genes including Adiponectin (Adipoq), fatty acid binding protein 4 (FABP4), and peroxisome proliferator-activated receptor γ (PPARγ). Results Compared with the control group, the body mass of the mice in the experimental group increased, the uterus atrophied, the bone mass decreased, the bone marrow fat expanded, and the bone metabolism showed a high bone turnover state. RT-qPCR showed that the expressions of cyp7a1, cyp8b1, and cyp27a1, which were related to the key enzymes of bile acid metabolism in the liver, decreased significantly (P<0.05), while the expression of cyp7b1 had no significant difference (P>0.05). Intervention with LCA at concentrations of 1, 10, and 100 μmol/L did not demonstrate any apparent toxic effects on BMSCs. Furthermore, LCA inhibited the expressions of osteogenic-related genes (ALP, Runx2, and OCN) in a dose-dependent manner, resulting in a reduction in ALP staining positive area. Concurrently, LCA promoted the expressions of adipogenic-related genes (Adipoq, FABP4, and PPARγ), and an increase in oil red O staining positive area. Conclusion After menopause, the metabolism of bile acids is altered, and secondary bile acid LCA interferes with the balance of osteogenic and adipogenic differentiation of BMSCs, thereby affecting bone remodelling.

Citation： WANG Cui, LI Jiao, LU Lingyun, LIU Lu, YU Xijie. Impact of lithocholic acid on the osteogenic and adipogenic differentiation balance of bone marrow mesenchymal stem cells. Chinese Journal of Reparative and Reconstructive Surgery, 2024, 38(1): 82-90. doi: 10.7507/1002-1892.202308050 Copy

1.	白璧辉, 谢兴文, 李鼎鹏, 等. 我国近5年来骨质疏松症流行病学研究现状. 中国骨质疏松杂志, 2018, 24(2): 253-258.
2.	Collins SL, Stine JG, Bisanz JE, et al. Bile acids and the gut microbiota: metabolic interactions and impacts on disease. Nat Rev Microbiol, 2023, 21(4): 236-247.
3.	Zhao YX, Song YW, Zhang L, et al. Association between bile acid metabolism and bone mineral density in postmenopausal women. Clinics (Sao Paulo), 2020, 75: e1486.
4.	Assis DN. Chronic complications of cholestasis: Evaluation and management. Clin Liver Dis, 2018, 22(3): 533-544.
5.	Han S, Li T, Ellis E, et al. A novel bile acid-activated vitamin D receptor signaling in human hepatocytes. Mol Endocrinol, 2010, 24(6): 1151-1164.
6.	Chaudhari SN, Luo JN, Harris DA, et al. A microbial metabolite remodels the gut-liver axis following bariatric surgery. Cell Host Microbe, 2021, 29(3): 408-424.
7.	Yao B, He J, Yin X, et al. The protective effect of lithocholic acid on the intestinal epithelial barrier is mediated by the vitamin D receptor via a SIRT1/Nrf2 and NF-κB dependent mechanism in Caco-2 cells. Toxicol Lett, 2019, 316: 109-118.
8.	Nguyen TT, Ung TT, Li S, et al. Lithocholic acid induces miR21, promoting PTEN inhibition via STAT3 and ERK-1/2 signaling in colorectal cancer cells. Int J Mol Sci, 2021, 22(19): 10209.
9.	Sheng W, Ji G, Zhang L. The effect of lithocholic acid on the gut-liver axis. Front Pharmacol, 2022, 13: 910493.
10.	Wang S, Wang S, Wang X, et al. Effects of icariin on modulating gut microbiota and regulating metabolite alterations to prevent bone loss in ovariectomized rat model. Front Endocrinol (Lausanne), 2022, 13: 874849.
11.	Hojo H, Ohba S. Gene regulatory landscape in osteoblast differentiation. Bone, 2020, 137: 115458.
12.	Li J, Chen X, Lu L, et al. The relationship between bone marrow adipose tissue and bone metabolism in postmenopausal osteoporosis. Cytokine Growth Factor Rev, 2020, 52: 88-98.
13.	Tencerova M, Ferencakova M, Kassem M. Bone marrow adipose tissue: Role in bone remodeling and energy metabolism. Best Pract Res Clin Endocrinol Metab, 2021, 35(4): 101545.
14.	Li Z, Hardij J, Bagchi DP, et al. Development, regulation, metabolism and function of bone marrow adipose tissues. Bone, 2018, 110: 134-140.
15.	Pachón-Peña G, Bredella MA. Bone marrow adipose tissue in metabolic health. Trends Endocrinol Metab, 2022, 33(6): 401-408.
16.	Guañabens N, Parés A, Ros I, et al. Severity of cholestasis and advanced histological stage but not menopausal status are the major risk factors for osteoporosis in primary biliary cirrhosis. J Hepatol, 2005, 42(4): 573-577.
17.	Parés A, Guañabens N, Rodés J. Gene polymorphisms as predictors of decreased bone mineral density and osteoporosis in primary biliary cirrhosis. Eur J Gastroenterol Hepatol, 2005, 17(3): 311-315.
18.	Yang S, Li H, Gu Y, et al. The association between total bile acid and bone mineral density among patients with type 2 diabetes. Front Endocrinol (Lausanne), 2023, 14: 1153205.
19.	Carson MD, Warner AJ, Hathaway-Schrader JD, et al. Minocycline-induced disruption of the intestinal FXR/FGF15 axis impairs osteogenesis in mice. JCI Insight, 2023, 8(1): e160578.
20.	Ma YS, Hou ZJ, Li Y, et al. Unveiling the pharmacological mechanisms of eleutheroside E against postmenopausal osteoporosis through UPLC-Q/TOF-MS-based metabolomics. Front Pharmacol, 2020, 11: 1316.
21.	Li Z, Huang J, Wang F, et al. Dual targeting of bile acid receptor-1 (TGR5) and farnesoid X receptor (FXR) prevents estrogen-dependent bone loss in mice. J Bone Miner Res, 2019, 34(4): 765-776.
22.	Id Boufker H, Lagneaux L, Fayyad-Kazan H, et al. Role of farnesoid X receptor (FXR) in the process of differentiation of bone marrow stromal cells into osteoblasts. Bone, 2011, 49(6): 1219-1231.
23.	Cho SW, An JH, Park H, et al. Positive regulation of osteogenesis by bile acid through FXR. J Bone Miner Res, 2013, 28(10): 2109-2121.
24.	Ahn TK, Kim KT, Joshi HP, et al. Therapeutic potential of tauroursodeoxycholic acid for the treatment of osteoporosis. Int J Mol Sci, 2020, 21(12): 4274.
25.	Ambrosi TH, Scialdone A, Graja A, et al. Adipocyte accumulation in the bone marrow during obesity and aging impairs stem cell-based hematopoietic and bone regeneration. Cell Stem Cell, 2017, 20(6): 771-784.
26.	Ruiz-Gaspà S, Guañabens N, Jurado S, et al. Bile acids and bilirubin effects on osteoblastic gene profile. Implications in the pathogenesis of osteoporosis in liver diseases. Gene, 2020, 725: 144167.
27.	Ruiz-Gaspà S, Guañabens N, Enjuanes A, et al. Lithocholic acid downregulates vitamin D effects in human osteoblasts. Eur J Clin Invest, 2010, 40(1): 25-34.
28.	Abdelkarim M, Caron S, Duhem C, et al. The farnesoid Ⅹ receptor regulates adipocyte differentiation and function by promoting peroxisome proliferator-activated receptor-gamma and interfering with the Wnt/beta-catenin pathways. J Biol Chem, 2010, 285(47): 36759-36767.

1. 白璧辉, 谢兴文, 李鼎鹏, 等. 我国近5年来骨质疏松症流行病学研究现状. 中国骨质疏松杂志, 2018, 24(2): 253-258.
2. Collins SL, Stine JG, Bisanz JE, et al. Bile acids and the gut microbiota: metabolic interactions and impacts on disease. Nat Rev Microbiol, 2023, 21(4): 236-247.
3. Zhao YX, Song YW, Zhang L, et al. Association between bile acid metabolism and bone mineral density in postmenopausal women. Clinics (Sao Paulo), 2020, 75: e1486.
4. Assis DN. Chronic complications of cholestasis: Evaluation and management. Clin Liver Dis, 2018, 22(3): 533-544.
5. Han S, Li T, Ellis E, et al. A novel bile acid-activated vitamin D receptor signaling in human hepatocytes. Mol Endocrinol, 2010, 24(6): 1151-1164.
6. Chaudhari SN, Luo JN, Harris DA, et al. A microbial metabolite remodels the gut-liver axis following bariatric surgery. Cell Host Microbe, 2021, 29(3): 408-424.
7. Yao B, He J, Yin X, et al. The protective effect of lithocholic acid on the intestinal epithelial barrier is mediated by the vitamin D receptor via a SIRT1/Nrf2 and NF-κB dependent mechanism in Caco-2 cells. Toxicol Lett, 2019, 316: 109-118.
8. Nguyen TT, Ung TT, Li S, et al. Lithocholic acid induces miR21, promoting PTEN inhibition via STAT3 and ERK-1/2 signaling in colorectal cancer cells. Int J Mol Sci, 2021, 22(19): 10209.
9. Sheng W, Ji G, Zhang L. The effect of lithocholic acid on the gut-liver axis. Front Pharmacol, 2022, 13: 910493.
10. Wang S, Wang S, Wang X, et al. Effects of icariin on modulating gut microbiota and regulating metabolite alterations to prevent bone loss in ovariectomized rat model. Front Endocrinol (Lausanne), 2022, 13: 874849.
11. Hojo H, Ohba S. Gene regulatory landscape in osteoblast differentiation. Bone, 2020, 137: 115458.
12. Li J, Chen X, Lu L, et al. The relationship between bone marrow adipose tissue and bone metabolism in postmenopausal osteoporosis. Cytokine Growth Factor Rev, 2020, 52: 88-98.
13. Tencerova M, Ferencakova M, Kassem M. Bone marrow adipose tissue: Role in bone remodeling and energy metabolism. Best Pract Res Clin Endocrinol Metab, 2021, 35(4): 101545.
14. Li Z, Hardij J, Bagchi DP, et al. Development, regulation, metabolism and function of bone marrow adipose tissues. Bone, 2018, 110: 134-140.
15. Pachón-Peña G, Bredella MA. Bone marrow adipose tissue in metabolic health. Trends Endocrinol Metab, 2022, 33(6): 401-408.
16. Guañabens N, Parés A, Ros I, et al. Severity of cholestasis and advanced histological stage but not menopausal status are the major risk factors for osteoporosis in primary biliary cirrhosis. J Hepatol, 2005, 42(4): 573-577.
17. Parés A, Guañabens N, Rodés J. Gene polymorphisms as predictors of decreased bone mineral density and osteoporosis in primary biliary cirrhosis. Eur J Gastroenterol Hepatol, 2005, 17(3): 311-315.
18. Yang S, Li H, Gu Y, et al. The association between total bile acid and bone mineral density among patients with type 2 diabetes. Front Endocrinol (Lausanne), 2023, 14: 1153205.
19. Carson MD, Warner AJ, Hathaway-Schrader JD, et al. Minocycline-induced disruption of the intestinal FXR/FGF15 axis impairs osteogenesis in mice. JCI Insight, 2023, 8(1): e160578.
20. Ma YS, Hou ZJ, Li Y, et al. Unveiling the pharmacological mechanisms of eleutheroside E against postmenopausal osteoporosis through UPLC-Q/TOF-MS-based metabolomics. Front Pharmacol, 2020, 11: 1316.
21. Li Z, Huang J, Wang F, et al. Dual targeting of bile acid receptor-1 (TGR5) and farnesoid X receptor (FXR) prevents estrogen-dependent bone loss in mice. J Bone Miner Res, 2019, 34(4): 765-776.
22. Id Boufker H, Lagneaux L, Fayyad-Kazan H, et al. Role of farnesoid X receptor (FXR) in the process of differentiation of bone marrow stromal cells into osteoblasts. Bone, 2011, 49(6): 1219-1231.
23. Cho SW, An JH, Park H, et al. Positive regulation of osteogenesis by bile acid through FXR. J Bone Miner Res, 2013, 28(10): 2109-2121.
24. Ahn TK, Kim KT, Joshi HP, et al. Therapeutic potential of tauroursodeoxycholic acid for the treatment of osteoporosis. Int J Mol Sci, 2020, 21(12): 4274.
25. Ambrosi TH, Scialdone A, Graja A, et al. Adipocyte accumulation in the bone marrow during obesity and aging impairs stem cell-based hematopoietic and bone regeneration. Cell Stem Cell, 2017, 20(6): 771-784.
26. Ruiz-Gaspà S, Guañabens N, Jurado S, et al. Bile acids and bilirubin effects on osteoblastic gene profile. Implications in the pathogenesis of osteoporosis in liver diseases. Gene, 2020, 725: 144167.
27. Ruiz-Gaspà S, Guañabens N, Enjuanes A, et al. Lithocholic acid downregulates vitamin D effects in human osteoblasts. Eur J Clin Invest, 2010, 40(1): 25-34.
28. Abdelkarim M, Caron S, Duhem C, et al. The farnesoid Ⅹ receptor regulates adipocyte differentiation and function by promoting peroxisome proliferator-activated receptor-gamma and interfering with the Wnt/beta-catenin pathways. J Biol Chem, 2010, 285(47): 36759-36767.

Chinese Journal of Reparative and Reconstructive Surgery

Impact of lithocholic acid on the osteogenic and adipogenic differentiation balance of bone marrow mesenchymal stem cells

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content