Astragalus Polysaccharides Alleviate Lung Adenocarcinoma Bone Metastases by Inhibiting the CaSR/PTHrP Signaling Pathway

Figure 1

APS inhibited RANKL-induced OCs differentiation and gene expression. (a, c, d) OC precursor cells were treated with different concentrations of APS or cultured for different time points to examine the potential effect on osteoclast differentiation. TRAP⁺ multinucleated cells were counted under a 400x microscope and the area of TRAP-stained osteoclasts was calculated with Image J software (NIH). The number of OCs and osteolytic area size in the APS-treated group were significantly lower than those of the negative control group (n = 3 per group). This inhibitory effect was concentration-dependent. (b, e, f) OCs formation was significantly inhibited 7 days after 200 mg/L of APS treatment according to TRAP staining (n = 3 per group). (g, h) The related protein expression of OCs differentiation cathepsin K and CTR, as well as NFATC1 and TRAP, were detected by Western blot after APS treatment. Image J software (NIH) for image processing was used to verify the band intensities as a result of semiquantitative western blots. In the APS treatment group, the protein expression levels of TRAP, cathepsin (K) CTR, and NFATC1 were significantly lower than those of the negative control group (n = 3 per group). The inhibitory effect was concentration-dependent. (, , , , the statistical test method was one-way ANOVA).

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