Effect of Angelica sinensis Polysaccharides on Osteoarthritis In Vivo and In Vitro: A Possible Mechanism to Promote Proteoglycans Synthesis
Figure 2
Effect of Angelica sinensis polysaccharides (APS-3c) on proteoglycan synthesis, cell proliferation, and cell apoptosis of rat interleukin-1-beta- (IL-1β-) stimulated chondrocytes. (a) Effect of APS-3c on glycosaminoglycan (GAG) contents in cell supernatants of rat IL-1β-stimulated chondrocytes. (b) Effect of APS-3c on proteoglycan synthesis of rat IL-1β-stimulated chondrocytes. Chondrocytes were incubated for the final 6 h with 10 μCi/mL 35S-sulfate in DMEM supplemented in the presence or absence of stimulants. After incubation, the media and cell extracts were incubated overnight at room temperature in 0.5% cetylpyridinium chloride. GAG was dissolved in Soluene-350 and the radioactivity of 35S-sulfate incorporated was measured by liquid scintillation counting. For each experiment, the amount of DNA was measured in sister flasks. Results were calculated as the mean ± SEM cell or medium disintegrations per minute/10 μg DNA in 6 similarly treated wells. (c) RT-PCR of aggrecan, Col2a1, and GAPDH (from top to bottom) mRNA in response to various concentrations of APS-3c. ((d) and (f)) The resulting data were expressed and illustrated as a ratio of the normal control. Integrated density values for polymerase products were normalized to the values for GAPDH. (e) Effect of APS-3c on apoptosis rate of rat IL-1β-stimulated chondrocytes. Chondrocytes were incubated with APS-3c alone for 4 h and then cotreated with IL-1β and APS-3c for 24 h. After being stained with annexin-V-fluoresceine isothiocyanate (FITC) and propidium iodide (PI), chondrocytes were measured on flow cytometry and analyzed with Multi-cycle software. (g) Effect of APS-3c on proliferation of rat chondrocytes in the presence or absence of IL-1β. Values represent Mean ± SEM of six different simples. versus normal control. , versus IL-1β-stimulated control.
