Glucoraphanin Increases Intracellular Hydrogen Sulfide (H2S) Levels and Stimulates Osteogenic Differentiation in Human Mesenchymal Stromal Cell

Abstract

Osteopenia and osteoporosis are among the most prevalent consequences of ageing, urging the promotion of healthy nutritional habits as a tool in preventing bone fractures. Glucosinolates (GLSs) are organosulfur compounds considered relatively inert precursors of reactive derivatives isothiocyanates (ITCs). Recent evidence suggests that GLSs may exert biological properties based on their capacity to release hydrogen sulfide (H2S). H2S-donors are known to exert anabolic function on bone cells. Here, we investigated whether a GLS, glucoraphanin (GRA) obtained from Tuscan black kale, promotes osteogenesis in human mesenchymal stromal cells (hMSCs). H2S release in buffer and intracellular H2S levels were detected by amperometric measurements and fluorimetric/cytofluorimetric analyses, respectively. Alizarin red staining assay and real-time PCR were performed to evaluate mineral apposition and mRNA expression of osteogenic genes. Using an in vitro cell culture model, our data demonstrate a sulforaphane (SFN)-independent osteogenic stimulation of GRA in hMSCs, at least partially attributable to H2S release. In particular, GRA upregulated the expression of osteogenic genes and enhanced mineral apposition while increasing intracellular concentrations of H2S. Overall, this study suggests the feasibility of using cruciferous derivatives as natural alternatives to chemical H2S-donors as adjuvant therapies in the treatment of bone-wasting diseases.

Beneficial Effect of H2S-Releasing Molecules in an In Vitro Model of Sarcopenia: Relevance of Glucoraphanin

“…All treatments were done in DM. Concentration and time of exposure for DEX-induced damage were previously set up [ 30 ]; glucoraphanin concentration was chosen based on the ability of the compound to increase the intracellular H 2 S concentration [ 84 ]; L-cysteine and 3-mercaptopyruvate concentrations were chosen based on previously obtained results [ 29 ].…”

Sulforaphane Inhibits IL-1β-Induced IL-6 by Suppressing ROS Production, AP-1, and STAT3 in Colorectal Cancer HT-29 Cells

“…The exploration of natural compounds and their analogs as chemopreventive agents is gaining momentum. SFN, known for its health benefits, is garnering attention in the medical field, especially for its potential protective effects against various types of cancer [38][39][40][41][42][43], cardiovascular diseases [44][45][46], neurological diseases [47][48][49][50], insulin resistance [51][52][53], obesity [54][55][56], and musculoskeletal diseases [57][58][59]. Additionally, the consumption of broccoli, brussels sprouts, cabbage, and SFN, an isothiocyanate derivative, is linked to various health benefits, including anticancer and antioxidant properties [60].…”

Dietary organosulfur compounds: Emerging players in the regulation of bone homeostasis by plant-derived molecules

“…(Continued) (97); at the molecular level, SFN induced up-regulation of RUNX-2 in mouse MSCs (97). Gambari et al showed increased mineralization and BSP, CBS and SMAD-1 mRNA up-regulation by GRA administration in primary human MSCs (98). Finally, with regards to osteocyte regulation, Thaler et al showed that SFN inhibited proliferation in murine osteocyte-like cell line (MLO-Y4) (97).…”

“…Gambari et al. showed increased mineralization and BSP, CBS and SMAD-1 mRNA up-regulation by GRA administration in primary human MSCs ( 98 ). Finally, with regards to osteocyte regulation, Thaler et al.…”

“…Of note, our group recently demonstrated that GRA obtained from Tuscan black kale promotes osteogenesis in human MSCs, independent of SFN, and this effect is associated to the release of H 2 S and an increased H 2 S uptake inside the cells ( 98 ). This is relatively unexpected as GLS have been considered for many years a relatively inert precursor of reactive derivatives ITCs.…”