Fig 1: Aorta for evidence of VSMC phenotype change by performing immunochemistry. Expression of CathepsinK (A-C), OPG (D-F), RANKL (G-I), TRAP (J-L), Runx2 (M-O), and Osteocalcin (P-R) were detected in the aortic tunica media of normal, CRF and 2%La treatment rats. Arrows indicate positively stained action. All sections were of the thoracic aorta region.
Fig 2: Evaluation of bone related markers in different groups by semi-quantitative scoring were demonstrated. 0: no expression; 1: focal expression; 2: partial expression; 3: circumferential expression. Immunohistochemical result showed that CathepsinK, RANKL and Osteocalcin were abundantly expressed whereas Runx2 was moderately expressed (p < 0.01) in CRF rats. Expression of Runx2, CathepsinK, RANKL and Osteocalcin were significantly down regulated in 2%La group (p < 0.01 vs CRF group). OPG were strongly positive in Control group and significantly down regulated in CRF group (p < 0.01 vs Control group) and up-regulated in 2%La group (p < 0.05 vs CRF group).
Fig 3: mRNA expressions of all factors relative to GAPDH were examined by qRT-PCR. Compared to control group, ## (p < 0.01); Compared to CRF Group, **(p < 0.01). mRNA expression of CathepsinK (A), RANKL (C), Runx2 (E) and Osteocalcin (F) were highly expressed (p < 0.01 vs control group) along with increased RANKL/OPG ratio (D) while OPG mRNA (B) was down-regulated in CRF group (p < 0.01 vs control). Binding of serum phosphate caused significantly decrease of CathepsinK, RANKL, Runx2 and Osteocalcin expression by 53.9%, 41.7%, 51.4% and 73.3% respectively (p < 0.01 vs CRF) whereas expression of OPG mRNA was found to be increased 1.7-fold (p < 0.01 vs CRF). The local RANKL/OPG ratio exhibited remarkable reduction in 2%La group (p < 0.01 vs CRF).
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