Hypoxia Inhibits Osteogenesis and Promotes Adipogenesis of Fibroblast-like Synoviocytes via Upregulation of Leptin in Patients with Rheumatoid Arthritis

Figure 5

Knockdown of leptin attenuated the hypoxia-mediated increase of adipogenesis of RA-FLSs under hypoxia conditions. (a) RT-qPCR analysis of leptin and adipogenesis-related genes (C/EBPα, PPARγ, and FABP4) in leptin-knockdown or scrambled siRNA control RA-FLSs after culture with adipogenic induction medium for 3 days under hypoxia and normoxia conditions. Data quantification is (-test, three independent experiments). (b, c) Westen blot analysis of leptin and adipogenesis-related proteins (C/EBPα and PPARγ) in leptin-knockdown or scrambled siRNA control RA-FLSs after culture with adipogenic induction medium for 3 days under hypoxia and normoxia conditions. The expression normalized with β-actin (-test, three independent experiments). (d) Oil Red O staining of leptin-knockdown or scramble siRNA control RA-FLSs after culture with adipogenic induction medium for 21 days under hypoxia and normoxia conditions. (e) Oil Red O positive cells per field were counted. Data quantification is . (-test, six randomized fields, three independent experiments, and one representative experiment shown). ; ; . Scale bars:

(a)

(b)

(c)

(d)

(e)