Crocin protects against dexamethasone‑induced osteoblast apoptosis by inhibiting the ROS/Ca2+‑mediated mitochondrial pathway

Abstract

Osteoblast apoptosis has been identified as an important event in the development of glucocorticoid (GC)-induced osteoporosis and osteonecrosis of the femoral head. Crocin, a bioactive ingredient of saffron, has been demonstrated to induce antiapoptotic effects on numerous types of cell in vitro ; however, the effects of crocin on the dexamethasone (Dex)-induced apoptosis of osteoblasts remain unclear. In the present study, the protective effects of crocin during Dex-induced apoptosis of MC3T3-E1 osteoblasts, and the underlying mechanisms, were investigated. MTT and Annexin V-FITC/PI flow cytometry assays were performed to evaluate the viability and apoptosis of cells, respectively. The mitochondrial transmembrane potential, reactive oxygen species (ROS), intracellular Ca 2+ levels and apoptosis-associated protein expression were assessed via flow cytometry, fluorescence microscopy and western blotting. It was demonstrated that crocin pretreatment inhibited Dex-induced apoptosis of osteoblasts in a dose-dependent manner. Crocin reversed Dex-induced decreases in the mitochondrial transmembrane potential, and increases in ROS and intracellular Ca 2+ levels. Furthermore, crocin upregulated the expression levels of B-cell lymphoma-2 (Bcl-2) and mitochondrial cytochrome c (Cyt C), and downregulated those of cleaved caspase-9, cleaved caspase-3, Bcl-2-associated X protein and cytoplasmic Cyt C. N-acetylcysteine, a ROS inhibitor, and 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid, a calcium chelator, attenuated Dex-induced osteoblast apoptosis, whereas H 2 O 2 and ionomycin, a calcium ionophore that increases intracellular calcium levels, reversed the antiapoptotic effects of crocin on Dex-treated osteoblasts. These results indicated that crocin may protect osteoblasts from Dex-induced apoptosis by inhibiting the ROS/Ca 2+ -mediated mitochondrial pathway, thus suggesting that crocin has potential value as a treatment for GC-induced bone diseases.

Crocin alleviates lipopolysaccharide-induced acute respiratory distress syndrome by protecting against glycocalyx damage and suppressing inflammatory signaling pathways

“…# p < 0.05 vs. control group, *p < 0.05 vs. LPS group participate in hemodynamics [5][6][7][8]25]. As shown in previous reports, crocin had an important protective effects in several diseases by affecting on cell apoptosis, arteriosclerosis, oxidation, and inflammation [15][16][17][18][19]. The present study demonstrated that pretreatment with crocin effectively improved the pulmonary edema associated with albumin leakage by maintaining the integrity of glycocalyx in LPS-induced ARDS mice.…”

“…Crocin is an ester glycoside extracted from saffron. As one of the main active constituents of saffron, crocin has many pharmacological effects, such as anti-oxidation [15], anti-arteriosclerosis [16], anti-apoptosis [17], and anti-inflammatory [18]. However, no studies have reported the effects of crocin on endothelial glycocalyx, HMGB-1, NF-κB, and MAPK signaling pathways in LPS-induced ARDS mice and LPS-stimulated HUVECs.…”

microRNA‐148a‐3p in extracellular vesicles derived from bone marrow mesenchymal stem cells suppresses SMURF1 to prevent osteonecrosis of femoral head

“…A previous study has demonstrated that osteoporosis and osteonecrosis are closely related to ONFH. 35 Especially, the inhibition of SMURF1 could improve the bone morphogenetic protein (BMP) signalling and the osteogenic differentiation while enhancing the capacity of bone formation in spinal fusion rats, and meanwhile SMURF1 could elevate the capacity of the bone fracture healing. 36,37 The knockdown of SMURF1 could promote the osteogenesis.…”

Effects of simvastatin on iNOS and caspase‑3 levels and oxidative stress following smoke inhalation injury

“…The levels of Mda, a product of the destruction of polyunsaturated fatty acids in the cell membrane, indirectly reflects the grade of oxidative damage. Sod acts as an important antioxidant enzyme by antagonizing and blocking oxygen free radicals to decrease cell damage and thus speeding the repair of free radical-induced damage in cells (54,55). High levels of oxygen free radicals exceed the capacity of the antioxidant system during smoke inhalation injury, which causes lipid peroxidation and subsequent destruction of cell membranes (56).…”