Journal of Shandong University (Health Sciences) ›› 2020, Vol. 58 ›› Issue (6): 14-21.doi: 10.6040/j.issn.1671-7554.0.2020.111

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Effects of medical ozone on the expression of PPARγ and autophagy in chondrocytes of osteoarthritis in rats

SUN Panpan1, ZHAO Xu2, LIN Xiaowen1, FU Zhijian1   

  1. 1. Department of Pain Management;
    2. Department of Anesthesia, Shandong Provincial Hospital Affiliated toShandong University, Jinan 250021, Shandong, China
  • Published:2022-09-27

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Abstract: Objective To investigate the effects of ozone(O3)on the expressions of peroxisome proliferators-activated receptor γ(PPARγ)and autophagy-related proteins in rat osteoarthritis chondrocytes and the role of PPARγ in ozone-induced autophagy of chondrocytes. Methods Wistar rats within 3 days of birth were chosen and the articular cartilage of extremities were obtained for the isolation and culture of primary chondrocytes. The chondrocytes were identified with toluidine blue staining and type II collagen immunofluorescence. The cells were stimulated with 10 ng/mL interleukin-1β(IL-1β)for 24 h to construct osteoarthritis chondrocytes models. After that, osteoarthritis chondrocytes were treated with different concentrations of ozone for 30 min, the effects on cell activity were detected by CCK-8 assay and the optimal O3 concentration of 30 μg/mL was selected. After IL-1β treatment, PPARγ-specific inhibitor GW9662 was added to the osteoarthritis chondrocytes and treated for 12 h, which were then treated with 30 μg/mL O3. The primary chondrocytes were randomly divided into control group, IL-1β group, IL-1β+O3 group, control+O3 group, IL-1β+O3+GW9662 group and IL-1β+GW9662 group. The expressions of PPARγ and autophagy-related proteins including LC3 Ⅱ, P62 and Beclin-1 were detected with Western blotting. The expressions of LC3B and P62 were detected with immunofluorescence staining. Results Toluidine blue staining and type II collagen immunofluorescence identified the cultured cells were chondrocytes. The cell activity was improved with 30 μg/mL ozone(P<0.05), and decreased with 50 and 70 μg/mL ozone(P<0.05). The 30 μg/mL ozone increased the expressions of PPARγ, LC3Ⅱ and Beclin-1, while decreased the level of P62(P<0.05). GW9662(20 mmol/mL)inhibited the up-regulation effect of ozone on the levels of LC3Ⅱ and Beclin-1, and reversed the inhibition of ozone on P62. Conclusion Ozone at 30 μg/mL can promote autophagy of osteoarthritis chondrocytes, and activation of PPARγ can promote ozone-induced autophagy in osteoarthritis chondrocytes.

Key words: Ozone, Interleukin-1β, Osteoarthritis, peroxisome proliferators-activated receptors, peroxisome proliferators-activated receptors, Autophagy

CLC Number: 

  • R575.1
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