Abstract
BackgroundOsteoclast cell increase is a major risk factor for osteoporosis and degenerative bone and joint diseases. At present, RANKL and M-CSF are commonly used to induce osteoclastogenesis. Thioacetamide (TAA) can lead to many types of liver and kidney damage, but less attention has been paid to the association of TAA with bone damage. In this work, we investigated the effects of TAA on the osteoclastogenesis and differentiation of bone marrow macrophages (BMMs).MethodsBMMs of SD rat suckling mice were taken for primary culture. CCK-8 was used to detect the toxic effects of TAA on BMMs, and flow cytometry was used to detect the effects of TAA on the cell cycle, cell viability, apoptosis and intracytoplasmic Ca2+ concentration of BMMs. TRAP staining was used to detect the effect of RANKL and M-CSF and TAA on osteoclast differentiation of BMMs. Western Blot was used to detect the expression level of PI3K/AKT pathway and osteoclast-specific proteins (TRAP and cathepsin K).ResultsThe results suggested that TAA inhibited the proliferation of BMMs, while enhancing osteoclastogenesis at 0.5 mg/mL and 1 mg/mL as assayed by TRAP staining. Exposed to TAA, BMMs could differentiate into osteoclast-like cells with overexpression of cathepsin K and TRAP proteins. Western blot results showed that TAA can activate the expression levels of P-PI3K, P-AKT, P-P38, and P-JNK, accompanied by apoptosis of BMMs and increase in intracellular Ca2+.ConclusionTAA may induce osteoclast formation in BMMs by activating the expression of PI3K/AKT pathway proteins, which is comparable to the classic osteoclast differentiation inducer RANKL and M-CSF. This suggests that we may find a cheap osteoclast inducer.
Highlights
TAA as a commercial chemical is widely used in electroplating additives, photographic drugs, pesticides and dyeing aids [1,2,3,4], and it is used to establish an animal model for liver cirrhosis due to its hepatotoxic effects [5]
Morphology and characterization of bone marrow macrophages (BMMs) culture in vitro We observed that freshly harvested BMMs were a uniform population; as cells continued to proliferate, the majority of cells became larger, and binuclear or even trinuclear can be seen
BMMs proliferation and viability were inhibited by TAAWe determined the inhibitory effect of TAA on BMMs using a CCK-8 assay upon exposure of BMMs
Summary
TAA as a commercial chemical is widely used in electroplating additives, photographic drugs, pesticides and dyeing aids [1,2,3,4], and it is used to establish an animal model for liver cirrhosis due to its hepatotoxic effects [5]. TAA-induced cirrhosis has similar pathologic changes to human liver cirrhosis and can be used to reproduce the human disease [6, 7]. The effects of TAA are not limited to the liver, as profound structural and functional changes have been described in the kidney, spleen, lung, intestine, stomach, brain and bone marrow after. Less attention has been paid to the association of TAA with bone damage. We investigated the effects of TAA on the osteoclastogenesis and differentiation of bone marrow macrophages (BMMs)
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