Mechanism of ykl-40 regulating apoptosis of rabbit osteoarthritis chondrocytes via pi3k/akt signaling pathway
10.3969/j.issn.2095-4344.2856
- Author:
Shenglan TIAN
1
Author Information
1. Wuhan University of Science and Technology Hospital
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Chondrocyte;
Experiment;
Human chitinase protein 40;
Knee;
Osteoarthritis;
Pathway;
Phosphatidylinositol 3-kinase
- From:
Chinese Journal of Tissue Engineering Research
2020;24(32):5108-5113
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Due to the close relationship between the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway and the physiological metabolism of bone tissue, human chitinase protein 40 (YKL-40) can regulate the PI3K/Akt signaling pathway related to breast cancer pathogenesis. Therefore, it is speculated that YKL-40 may regulate apoptosis in knee osteoarthritis (KOA) chondrocytes through the PI3K/Akt signaling pathway. OBJECTIVE: To study the mechanism by which YKL-40 regulates apoptosis in rabbit KOA chondrocyte s through the PI3K/Akt signaling pathway. METHODS: (1) New Zealand white rabbits were randomized into two groups. An animal model of KOA was made using anterior cruciate ligament dissection in the model group, whereas the right posterior knee joint capsule was cut but not dissected in the control group. Chondrocytes were extracted from the rabbits at 6 weeks after modeling. Hematoxylin-eosin staining and Mankin histological scoring of the cartilage tissue were performed, whereas immunohistochemical staining was used to detect type II collagen expression in chondrocytes. (2) The second-generation chondrocytes in the control group were used as normal control group, and those in the model group were further divided into four groups, followed by culture with high glucose DMEM medium containing 10% fetal bovine serum in KOA model group, 100 μg/L YKL-40 in KOA + YKL group, 50 µmol/L LY294002 in KOA + LY group, and 50 µmol/L LY294002 + 100 μg/L YKL-40 in KOA + YKL + LY group. The expression levels of collagen type II, matrix metalloproteinase 13, Akt, p-Akt, P53, Bcl-2 proteins in chondrocytes were detected by western blot. RESULTS AND CONCLUSION: Hematoxylin-eosin staining, Mankin histological score, and collagen type II immunohistochemical staining confirmed the successful construction of KOA animal model and successful chondrocyte culture. Compared with the normal control group, the collagen type II, Bcl-2, p-Akt protein expression levels in chondrocytes were significantly reduced in the KOA model group (P < 0.05), and matrix metalloproteinase 13 and P53 protein expression levels were significantly increased in the KOA model group (P < 0.05). Compared with the KOA model group, these indicators were significantly improved in the KOA + YKL group (P < 0.05), significantly worsened in the KOA + LY group (P < 0.05), and had no significant changes in the KOA + YKL + LY group (P > 0.05). However, there were significant differences between the KOA + YKL group and the KOA + LY group (P < 0.05). In addition, the expression level of p-Akt protein in chondrocytes had no difference among groups (P > 0.05). To conclude, YKL-40 can inhibit the apoptosis of KOA chondrocytes, accelerate the repair of KOA cartilage damage and delay the degeneration of cartilage by activating the PI3K/Akt signaling pathway. Therefore, YKL-40 can be used as a new target for clinical treatment of KOA.
