Long noncoding RNA ZFAS1 suppresses chondrocytes apoptosis via miR-302d-3p/SMAD2 in osteoarthritis.

Osteoarthritis (OA) seriously affects people's quality of life due to joint pain, stiffness, disability, and dyskinesia worldwide. Long noncoding RNA zinc finger antisense 1 (ZFAS1) is downregulated and tightly associated with proliferation, migration, apoptosis, and matrix synthesis of chondrocyte in OA. However, the molecular mechanisms of ZFAS1 in OA remain unknown. The expression correlation between ZFAS1, miR-302d-3p, and SMAD2 in OA tissues was analyzed by Pearson correlation analysis. ZFAS1 was a lower expression, and expedited proliferation and repressed apoptosis of chondrocytes. MiR-302d-3p was a direct target of ZFAS1. MiR-302d-3p hindered proliferation and facilitated apoptosis of chondrocytes. MiR-302d-3p partially reversed the effect of ZFAS1 on proliferation and apoptosis of chondrocytes. SMAD2 was positively regulated by the ZFAS1/miR-302d-3p. MiR-302d-3p-mediated proliferation and apoptosis were partly abrogated by targeting SMAD2. ZFAS1 promoted chondrocytes proliferation and repressed apoptosis possibly by regulating miR-302d-3p/SMAD2 axis, providing a potential target for OA treatment.

The molecular mechanism of treating osteoarthritis with dipsacus saponins by inhibiting chondrocyte apoptosis.

The present study aimed to determine the molecular mechanism of treating osteoarthritis with dipsacus saponins by inhibiting the apoptosis of chondrocytes. A total of 30 New Zealand rabbits were randomly divided into 2 groups: A control group and a model group. The osteoarthritis model was established using the HULTH method. The success of the model establishment was determined by pathological morphology and measurement of inflammatory cytokine levels. Chondrocytes were isolated and divided into 4 groups treated with varying concentrations of dipsacus saponins: 0, 25, 50 and l00 µg/l dipsacus saponins. Cell cycle distribution was analyzed using flow cytometry. Changes in cyclin D1, cyclin-dependent kinase 4 (CDK4) and p21 expression were detected by western blotting and changes in the levels of Bcl-2, Bax, caspase-3 and caspase-9 mRNA were detected using reverse transcription-quantitative polymerase chain reaction. The osteoarthritis model was established successfully, indicated by a significant increase in the levels of IL-1ß, IL-6 and TNF-α in the model group (P<0.05) compared with the control group. The viability of the chondrocytes increased following treatment with dipsacus saponins in a concentration-dependent manner. The number of chondrocytes in the G0/G1 phase decreased in the 50 and l00 µg/l groups while the number of chondrocytes in the G2/M phase increased in the 50 and l00 µg/l groups. Levels of cyclin D1 and CDK4 expression increased following treatment with dipsacus saponins. Levels of Bax, caspase-3 and caspase-9 expression decreased while Bcl-2 levels increased following treatment with dipsacus saponins. The viability of chondrocytes increased following treatment with dipsacus saponins in a concentration-dependent manner. Thus, dipsacus saponins inhibited the apoptosis of chondrocytes by up-regulating Bcl-2 and down-regulating caspase-9, caspase-3 and Bax expression.

[The effect of fibroblast growth factors and epidermal growth factors on the proliferation of rabbit anterior cruciate and medial collateral ligament cell in vitro].

OBJECTIVE: To investigate the effect of acid, basic fibroblast, growth factor (aFGF, bFGF) and epidermal growth factor (EGF), and their combination on the proliferation of rabbit anterior cruciate ligament (ACL) and medial collateral ligament (MCL) in vitro. METHODS: The cells of ACL and MCL were isolated and subcultured from the knee joints of ten-week-old New Zealand white rabbits. The cells were seeded into 96-well corning cluster plates. Three growth factors of different concentration alone or in combination were added into the culture medium respectively, which were 0, 1, 5, 10, 50 and 100 ng/ml for aFGF, bFGF and 0, 1.56, 3.13, 6.25, 12.5, 25 and 50 ng/ml for EGF. The proliferation of the fibroblasts was measured for 48 h with XTT method. RESULTS: All of the three growth factors alone promoted the cell proliferation of ACL and MCL fibroblasts. The concentration of aFGF had a significant effect on the proliferation of both ACL and MCL fibroblasts. The concentration of 1 ng/ml bFGF and 5 ng/ ml EGF was most effective in promoting the proliferation of ACL, and both bFGF and EGF had a significant effect on MCL. 5 ng/ml aFGF with 50 ng/ml EGF had effect on ACL. 1 ng/ml aFGF with 3.13 ng/ml EGF had effect on MCL. CONCLUSION: The three growth factors may promote the cell proliferation of ACL and MCL. These findings suggest that topical application of aFGF, either alone or in combination with EGF may have the potential to promote the proliferation of rabbit ACL and MCL,and aFGF of low concentration in combination with EGF is more effective than single growth factor.