Resveratrol induces proliferation and differentiation of mouse pre-osteoblast MC3T3-E1 by promoting autophagy

Abstract

Background In mouse, it was discovered that resveratrol (Res) enhanced osteoporosis (OP) by boosting osteogenesis. Besides, Res can also have an impact on MC3T3-E1 cells, which are crucial for the control of osteogenesis and thus increase osteogenesis. Although some articles have discovered that Res enhanced autophagy to promote the value-added differentiation of MC3T3, it is unclear exactly how this affects the process of osteogenesis in mouse. Therefore, we will show that Res encourages MC3T3-E1 proliferation and differentiation in mouse pre-osteoblasts and further investigate the autophagy-related mechanism for this impact. Methods (1) MC3T3-E1 cells were separated into blank control group and various concentrations (0.01, 0.1, 1, 10, 100µmol/L) of group in order to determine the ideal Res concentration. In the Res group, Cell Counting Kit-8 (CCK-8) was used to measure the proliferation activity of pre-osteoblasts in mice in each group after resveratrol intervention. Alkaline Phosphatase (ALP) and alizarin red staining were used to gauge the degree of osteogenic differentiation, and RT-qPCR was used to measure the expression levels of Runx2 and OCN in the osteogenic differentiation ability of the cells. (2) In the experiment, four groups were set up: the control group, 3MA group, Res group, and Res + 3MA group. To examine cell mineralization, ALP and alizarin red staining were utilized. RT-qPCR and Western blot detection of cell autophagy activity levels and osteogenic differentiation capacity in each group following intervention. Results (1) Resveratrol might increase the number of mice pre-osteoblast, with the impact being most pronounced at 10µmol/L (P < 0.05). The nodules developed substantially more often than in the blank control group, and Runx2 and OCN expressions significantly increased (P < 0.05). (2) In contrast to the Res group, after 3MA purine blocked autophagy, the Res + 3MA group’s alkaline phosphatase staining and the development of mineralized nodules were reduced. Runx2, OCN, LC3II / LC3I expression decreased, p62 expression increased (P < 0.05). Conclusion The present study partially or indirectly demonstrated that Res may, through increased autophagy, induce osteogenic differentiation of MC3T3-E1 cells.

Prevention and Co-Management of Breast Cancer-Related Osteoporosis Using Resveratrol

“…Moreover, results from a rat model with postmenopausal OP have demonstrated that resveratrol increases autophagy, thereby promoting osteoblast differentiation [ 191 ]. These findings are in accordance with observations suggesting that the number of mouse pre-osteoblasts rises under resveratrol treatment through increased autophagy [ 190 ], which can be interpreted as protection against epigenetic changes. Furthermore, in vivo results demonstrated a protective role of the natural polyphenol on osteoblasts by increasing the expression of Beclin-1 and LC3 through up-regulation of Sirt-1, associated with suppression of the mTOR/Akt/PI3K pathway [ 192 ].…”

“…A beneficial characteristic of resveratrol consists in its modulation of processes such as autophagy and apoptosis in osteoblasts and MSC under osteoporotic microenvironments in in vitro and in vivo OP models, promoting cell survival, function, and differentiation, which is crucial for the regeneration of ECM in bone tissue [ 190 ] ( Figure 7 ). Moreover, results from a rat model with postmenopausal OP have demonstrated that resveratrol increases autophagy, thereby promoting osteoblast differentiation [ 191 ].…”