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Updated: Dec 30, 2025

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro
Published on: June 16, 2022
Protosappanin a inhibits osteoclastogenesis via reducing oxidative stress in RAW264.7 cells
Qiang Huang1, Rui-Ying Meng1, Yan-Wei Yang1
1Lanzhou General Hospital of Lanzhou Military Command Lanzhou, Gansu, People's Republic of China.
Abstract:
Protosappanin A (PrA), obtained from the traditional Chinese herbal medicine, Caesalpinia sappan L. (Lignum Sappan), possesses a lot of pharmaceutical activities. Typically, it is a potent antioxidant. This study makes an effort to test its protective effects against osteoporosis by partially reducing oxidative stress in RAW264.7 cells and a mouse ovariectomized (OVX) osteoporosis model. The influence that PrA affected on osteoclastic proliferation and differentiation under oxidative status was investigated. Our results revealed that PrA significantly inhibited the proliferation of RAW264.7 cells in oxidative stress conditions. Moreover, it suppressed some osteoclastic markers by TRAP staining, bone section assay and quantitative real-time PCR. PrA decreased reactive oxygen species (ROS) generation in RAW264.7 cells. In vivo, our results demonstrated that PrA supplementation improved some serum oxidative markers, including malondialdehyde (MDA) and reduced glutathione (GSH), and inhibited some osteoclastic markers, such as CTX-1 and TRAP. Importantly, it ameliorated the micro-architecture of trabecular bones by micro-CT assay. In summary, these findings showed that protection by PrA against osteoporosis is associated with a reduction in oxidative stress, suggesting that PrA may be useful in bone resorption related diseases, especially osteoporosis.
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