LI Lin, ZHU Biao, TIAN Xiaoyu, DANG Ruijie, ZHAO Lisheng, YANG Shuo, SUN Lei, WEN Ning. Effect of cannabidiol on osteogenic differentiation of senescent mice bone marrow mesenchymal stem cells in vitro[J]. ACADEMIC JOURNAL OF CHINESE PLA MEDICAL SCHOOL, 2022, 43(1): 75-80, 120. DOI: 10.3969/j.issn.2095-5227.2022.01.015
Citation: LI Lin, ZHU Biao, TIAN Xiaoyu, DANG Ruijie, ZHAO Lisheng, YANG Shuo, SUN Lei, WEN Ning. Effect of cannabidiol on osteogenic differentiation of senescent mice bone marrow mesenchymal stem cells in vitro[J]. ACADEMIC JOURNAL OF CHINESE PLA MEDICAL SCHOOL, 2022, 43(1): 75-80, 120. DOI: 10.3969/j.issn.2095-5227.2022.01.015

Effect of cannabidiol on osteogenic differentiation of senescent mice bone marrow mesenchymal stem cells in vitro

  •   Background  Bone marrow mesenchymal stem cells (BMSCs) in senescent state have a reduced capacity for osteogenic differentiation, which hazards to normal bone metabolism. Autophagy plays an important role in cellular senescence process and osteogenic metabolism. Cannabidiol (CBD) activates autophagy, but whether CBD can promote osteogenic differentiation of senescent BMSCs via regulating autophagy remains unclear.
      Objective  To investigate the effect of CBD on osteogenic differentiation of senescent mice BMSCs and the role of cellular autophagy.
      Methods  Mice BMSCs were resuscitated and cultured. D-galactose (D-gal) was used to induce an in vitro senescence model. Cells were randomly divided into control (Vehicle) group, senescence model (D-gal) group and CBD intervention (D-gal+CBD) group. qRT-PCR was performed to detect the mRNA expression levels of senescence-related genes, and CCK-8 assay was used to detect BMSCs proliferation activity. BMSCs in autophagy inhibition group (D-gal+CBD+Chloroquine) were added, and after osteogenesis induction of the four cells groups, the expression levels of autophagy-related proteins, P62 and LC3B, were detected by Western blot. qRT-PCR was performed to assess the mRNA expression levels of osteogenesis-related genes, and ALP activity was determined by alkaline phosphatase (ALP) assay kit.
      Results  BMSCs treated with D-gal overexpressed senescence-related genes P16 and P21, and exhibited decreased cell proliferation activity. The protein expression level of P62 increased while the protein expression level of LC3B-Ⅱ reduced, and the mRNA expression levels of ALP, Runx2, OCN and ALP activity were also decreased (P<0.05); In contrast, CBD intervention down-regulated the mRNA expression levels of P16 and P21 in senescent BMSCs. CBD significantly decreased P62 and increased the LC3B-Ⅱ protein levels, promoted the mRNA expression levels of ALP, Runx2 and OCN, and significantly enhanced ALP activity (P<0.05). In addition, the activation of autophagy and osteogenic differentiation of senescent BMSCs promoted by CBD were weakened by the autophagy inhibitor chloroquine (P<0.05).
      Conclusion  CBD promotes osteogenic differentiation of senescent mice BMSCs via activating autophagy.
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