Tanshinol stimulates bone formation and attenuates dexamethasone-induced inhibition of osteogenesis in larval zebrafish

Abstract

SummaryBackground/ObjectiveTanshinol is the main active component of Salvia miltiorrhiza Bunge, a significant Traditional Chinese Medicine used to treat cardiovascular disease. We have shown that tanshinol exerts an antiosteoporostic effect via the enhancement of bone formation in vivo and in vitro. However, the mechanism remains unclear. Based on the polyphenol group in the structure of tanshinol, we speculate the protective action on skeletal tissue is related to antioxidative capacity. Our in vitro evidence indicated that tanshinol stimulated osteoblastic differentiation by its antioxidaive capacity. In this study, we aim to further confirm the effect of tanshinol on bone formation and the underlying mechanism in zebrafish in vivo.MethodsWe used a Danio rerio (zebrafish) model, which has a bone formation process similar to humans, and evaluated the relationship between the dose and the effect of tanshinol on bone formation determined using alizarin red S staining or fluorescence intensity analysis in normal and glucocorticoid (GC)-induced inhibition of an osteogenesis model using wild-type zebrafish and cortical bone transgenic zebrafish tg(sp7:egfp). The expression of osteoblast-specific genes and reactive oxygen species (ROS) were tested.ResultsOur data showed that dexamethasone exerts a series of consequences, including the inhibition of bone formation, decrease of bone mass, downregulation of expression of osteoblast-specific genes (runx2a, ALP, osteocalcin, and sp7), as well as the accumulation of ROS generation and decreased capacity of antioxidants. Tanshinol showed a protective effect on promoting bone formation and bone mass both in wild-type larval zebrafish and transgenic zebrafish. Furthermore, tanshinol attenuated the inhibition of osteogenesis elicited by oxidative stress in the zebrafish exposed to dexamethasone.ConclusionThe present findings suggest that tanshinol prevented decreased osteogenesis in GC-treated larval zebrafish via scavenging ROS and stimulated the expression of osteoblast-specific genes. Tanshinol treatment may be developed as a novel therapeutic approach under recent recognised conditions of GC-induced osteoporosis.

The zebrafish operculum: A powerful system to assess osteogenic bioactivities of molecules with pharmacological and toxicological relevance

“…Several studies have reported the use of alizarin red S to screen for molecules with osteogenic and mineralogenic bioactivities in zebrafish. They assessed the mineralization of cranial bones in larvae exposed from 3 to 9 dpf (Luo et al, 2016), or the count of vertebral bodies at 10 dpf (Yu et al, 2008), but none considered inter-specimen variability or accurately quantified the osteogenic effect. They also failed to provide a medium/high-throughput approach, utilised long exposures, late endpoints and a laborious image acquisition procedure, and therefore have limited applicability for large-scale screening of molecules.…”

<p>Baicalin Ameliorates Dexamethasone-Induced Osteoporosis by Regulation of the RANK/RANKL/OPG Signaling Pathway</p>

“…Zebrafish is an ideal animal model in vivo for studying bone deformations for the high similarity of the structure and genetics of bone with human beings. 34,44 The bone of zebrafish larval contains the sufficient and necessary cells for both bone formation and resorption activity. 33 In the present study, we examined the bone mineralization by calcein staining, which is a vital dye staining and the biological mineralization process of zebrafish at phenotypic can be observed directly.…”

“…Dexamethasone (DEX)-induced osteopenia zebrafish larvae has been proved to be a well-established animal model for study of osteoporosis. 34 In this study, zebrafish larvae at 96 hpf (hours post fertilization) were exposed to DEX solutions with a concentration of 25 μM/L (DEX+), and 0.01% DMSO (DEX-) was selected as the vehicle control. Etidronate disodium (ED) is known to reduce bone resorption through the inhibition of osteoclastic activity.…”

Rolling Bearing Fault Diagnosis Using Modified Neighborhood Preserving Embedding and Maximal Overlap Discrete Wavelet Packet Transform with Sensitive Features Selection

“…WT can be treated as a fast-evolving mathematical and signal processing tool in dealing with nonstationary signals [66] and has been widely applied in many engineering fields for decomposing, denoising, and signal analysis over nonstationary signals [26,42]. Continuous wavelet transform (CWT) and discrete wavelet transform (DWT) are two categories of WT.…”

“…EMD has some problems such as over envelope, end effects, and mode mixing [22][23][24]. The effectiveness of STFT is still hampered by the limitation of single triangular basis [25,26]. WVD can produce interference terms on the time-frequency domain in a critical condition and high computational complexity [27].…”