Shikonin stimulates MC3T3-E1 cell proliferation and differentiation via the BMP-2/Smad5 signal transduction pathway

Abstract

Abstract. Shikonin, the predominant naphthoquinone pigment isolated from the Chinese plant Lithospermum erythrorhizon, is anti-inflammatory, antiviral and exerts anticancer effects, amongst other biological activities. However, it is unknown whether shikonin affects bone formation. In the present study, the role of shikonin on cell proliferation was assessed via MTT assay, and shikonin was identified to markedly promote cell growth in a time-and dose-dependent manner in the MC3T3-E1 cell line. In addition, flow cytometric analysis was performed to evaluate the effect of shikonin on the cell cycle, and it was observed that shikonin markedly increased the percentage of S-phase MC3T3-E1 cells to accelerate the G 1 /S transition. To investigate the potential molecular mechanism by which shikonin enhances bone formation, the changes in bone morphogenic protein-2 (BMP-2), SMAD family member 5 (Smad5), runt related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and osteocalcin (OC) expression levels induced by shikonin were investigated using western blot analysis and quantitative polymerase chain reaction. The results indicated that shikonin increased the BMP-2 and Smad5 mRNA levels, and upregulated Smad5 and Runx2 protein expression levels to promote osteoblast differentiation. Furthermore, ALP staining was performed, and revealed that shikonin enhanced ALP activity. These results indicate that shikonin promotes cell proliferation and differentiation of MC3T3-E1 cells via the BMP-2/Smad5 signaling pathway.

Combined antibacterial and osteogenic in situ effects of a bifunctional titanium alloy with nanoscale hydroxyapatite coating

“…This result corroborates the wide-held belief that porous structure and the calcium component of nano-HA can accelerate osteoblast proliferation [41][42][43][44]. Additionally, we were able to confirm that HBD-3 and BMP-2 do not affect the cells' proliferation behaviour, which was in agreement with previous studies [45][46][47].…”

Pseudoshikonin I enhances osteoblast differentiation by stimulating Runx2 and Osterix

“…31 Additionally, shikonin, the predominant naphthoquinone pigment of Lithospermum erythrorhizon was shown to stimulate differentiation of MC3T3-E1 pre-osteoblastic cells throuth BMP-2/Smad5 signal pathway. 43 Moreover, administration of shikonin in adjuvant-induced arthritic mice prevented reductions in bone mass density and trabecular bone volume. 29 PSI, new bioactive constituent of Lithospermi radix, which has similar chemical structure compared with shikonin, was found to inhibit matrix metalloproteinases by targeting IL-1β.…”

“…Recently, the ethanol (EtOH) extracts of LES were demonstrated to influence the functions of Osterix and Runx2 in osteoblastogenesis . Additionally, shikonin, the predominant naphthoquinone pigment of Lithospermum erythrorhizon was shown to stimulate differentiation of MC3T3‐E1 pre‐osteoblastic cells throuth BMP‐2/Smad5 signal pathway . Moreover, administration of shikonin in adjuvant‐induced arthritic mice prevented reductions in bone mass density and trabecular bone volume .…”

Long noncoding RNA SMAD5‐AS1 acts as a microRNA‐106a‐5p sponge to promote epithelial mesenchymal transition in nasopharyngeal carcinoma

“…SMAD5 is a fundamental component of heterometric SMAD complex, which often localizes into the nucleus and regulates gene transcription (16). SMAD5 has been proved to be involved in the process of osteoblast proliferation and differentiation (17). Moreover, miR-27a and miR-23a contribute to enhanced apoptosis in human granulosa cells through regulating SMAD5 (18).…”