Abstract
Background Cryptotanshinone (CPT), an active component extracted from the root of Salvia miltiorrhiza Bunge, exhibits extensive favorable bioactive properties including anti-inflammatory, antioxidative, antibacterial, and antitumor effects. This study aims to investigate the effects of CPT on osteogenesis and explore related mechanisms both in vivo and in vitro. Methods In the in vivo experiment, ovariectomized (OVX) female rats were intragastrically administered with CPT at doses of 10 mg/kg and 20 mg/kg for 13 consecutive weeks. Dual-energy X-ray absorptiometry was employed to detect bone mineral density (BMD). ELISA assay was leveraged to detect the biochemical parameters such as BUN and creatinine in the kidney samples. Bone and kidney sections were dyed by H&E and Masson staining kits. In the in vitro experiment, the RAW 264.7 cells were stimulated through the receptor activation of the nuclear factor kappa B ligand (RANKL) to establish an osteoclast differentiation model, and CPT's protective effect against bone loss was evaluated. Differentiated osteoclasts were determined by TRAP staining. While, osteoclast-marker proteins such as NFATc1, c-Fos, and cathepsin K were identified by Western blot. Results The results from in vivo experiments revealed that CPT could elevate bone mass and increase bone formation markers in OVX rats. Intriguingly, CPT administration noticeably ameliorated the kidney injury in OVX rats by suppressing BUN and restoring creatinine levels. Furthermore, the results from in vitro experiments suggested that CPT downregulated the protein expression of osteoclast-associated genes such as cathepsin K, c-Fos, and NFATc1 which hinted the related potential mechanisms. Conclusion The evidence from in vivo and in vitro experiments suggested that CPT exerted antiosteoclastogenic effects by inhibiting the activation of osteoclast differentiation followed by suppressing the protein expressions of cathepsin K, c-Fos, and NFATc1 in osteoclast precursors, and it exhibited protective effects against kidney damage, which highlighted its advantage in clinical application.
Highlights
Cryptotanshinone (CPT), an active component extracted from the root of Salvia miltiorrhiza Bunge, exhibits extensive favorable bioactive properties including anti-inflammatory, antioxidative, antibacterial, and antitumor effects. is study aims to investigate the effects of CPT on osteogenesis and explore related mechanisms both in vivo and in vitro
CPT was purchased from Dongguan Hanshuo Jianyuan Biotechnology (Shenzhen, China), while RANKL was purchased from R&D Systems (Minneapolis, MN, USA) and dissolved in phosphate-buffered saline. e ELISA kits, the detection of blood urea nitrogen (BUN), creatinine, ALP, OPG, and RANKL were purchased from Wuhan Huamei Biological Co., Ltd. (Wuhan, China)
JYB1-1 calcium removal solution and protein extraction kit were purchased from Solarbio Company (Beijing, China). e tartrate-resistant acid phosphatase (TRAP) staining kit was obtained from Whatman (England), and the MTT assay kit was purchased from Abcam (England)
Summary
Cryptotanshinone (CPT), an active component extracted from the root of Salvia miltiorrhiza Bunge, exhibits extensive favorable bioactive properties including anti-inflammatory, antioxidative, antibacterial, and antitumor effects. is study aims to investigate the effects of CPT on osteogenesis and explore related mechanisms both in vivo and in vitro. E results from in vivo experiments revealed that CPT could elevate bone mass and increase bone formation markers in OVX rats. The results from in vitro experiments suggested that CPT downregulated the protein expression of osteoclastassociated genes such as cathepsin K, c-Fos, and NFATc1 which hinted the related potential mechanisms. E evidence from in vivo and in vitro experiments suggested that CPT exerted antiosteoclastogenic effects by inhibiting the activation of osteoclast differentiation followed by suppressing the protein expressions of cathepsin K, c-Fos, and NFATc1 in osteoclast precursors, and it exhibited protective effects against kidney damage, which highlighted its advantage in clinical application. As estrogen regulates factors associated with the balance of bone formation and absorption [6], postmenopausal women are at high risk of developing osteoporosis owing to estrogen deficiency. The most common therapy for the prevention and treatment of postmenopausal osteoporosis [9, 10], has long been evidenced to be correlated with elevated risks of breast cancer, ovarian cancer, endometrial cancer, and cardiovascular diseases in postmenopausal women [11, 12]. erefore, estrogen therapy is not the optimal option to prevent fractures in postmenopausal women, and it is imperative to creatively develop effective and efficient treatment strategies in this regard
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