Effects of VX765 on osteoarthritis and chondrocyte inflammation in rats.
10.7507/1002-1892.202308053
- Author:
Wanran HUANG
1
;
Junxue TU
1
;
Aiqing QIAO
1
;
Chujun HE
1
Author Information
1. Department of Pharmacy, Yuying Children's Hospital, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou Zhejiang, 325000, P. R. China.
- Publication Type:Journal Article
- Keywords:
VX765;
chondrocytes;
nuclear factor erythroid 2-related factor 2 pathway;
osteoarthritis;
rat
- MeSH:
Rats;
Animals;
Chondrocytes/metabolism*;
Matrix Metalloproteinase 13/metabolism*;
Rats, Sprague-Dawley;
Tumor Necrosis Factor-alpha/metabolism*;
Collagen Type II/metabolism*;
Interleukin-6;
Lipopolysaccharides/pharmacology*;
NF-E2-Related Factor 2/pharmacology*;
Inflammation/drug therapy*;
Osteoarthritis/metabolism*;
Transforming Growth Factor beta1/metabolism*;
Dipeptides;
para-Aminobenzoates
- From:
Chinese Journal of Reparative and Reconstructive Surgery
2024;38(1):74-81
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effects and underlying mechanisms of VX765 on osteoarthritis (OA) and chondrocytes inflammation in rats.
METHODS:Chondrocytes were isolated from the knee joints of 4-week-old Sprague Dawley (SD) rats. The third-generation cells were subjected to cell counting kit 8 (CCK-8) analysis to assess the impact of various concentrations (0, 1, 5, 10, 20, 50, 100 μmol/L) of VX765 on rat chondrocyte activity. An in vitro lipopolysaccharide (LPS) induced cell inflammation model was employed, dividing cells into control group, LPS group, VX765 concentration 1 group and VX765 concentration 2 group without obvious cytotoxicity. Western blot, real-time fluorescence quantitative PCR, and ELISA were conducted to measure the expression levels of inflammatory factors-transforming growth factor β 1 (TGF-β 1), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α). Additionally, Western blot and immunofluorescence staining were employed to assess the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1). Thirty-two SD rats were randomly assigned to sham surgery group (group A), OA group (group B), OA+VX765 (50 mg/kg) group (group C), and OA+VX765 (100 mg/kg) group (group D), with 8 rats in each group. Group A underwent a sham operation with a medial incision, while groups B to D underwent additional transverse incisions to the medial collateral ligament and anterior cruciate ligament, with removal of the medial meniscus. One week post-surgery, groups C and D were orally administered 50 mg/kg and 100 mg/kg VX765, respectively, while groups A and B received an equivalent volume of saline. Histopathological examination using HE and safranin-fast green staining was performed, and Mankin scoring was utilized for evaluation. Immunohistochemical staining technique was employed to analyze the expressions of matrix metalloproteinase 13 (MMP-13) and collagen type Ⅱ.
RESULTS:The CCK-8 assay indicated a significant decrease in cell viability at VX765 concentrations exceeding 10 μmol/L ( P<0.05), so 4 μmol/L and 8 μmol/L VX765 without obvious cytotoxicity were selected for subsequent experiments. Following LPS induction, the expressions of TGF-β 1, IL-6, and TNF-α in cells significantly increased when compared with the control group ( P<0.05). However, intervention with 4 μmol/L and 8 μmol/L VX765 led to a significant decrease in expression compared to the LPS group ( P<0.05). Western blot and immunofluorescence staining demonstrated a significant upregulation of Nrf2 pathway-related molecules Nrf2 and HO-1 protein expressions by VX765 ( P<0.05), indicating Nrf2 pathway activation. Histopathological examination of rat knee joint tissues and immunohistochemical staining revealed that, compared to group B, treatment with VX765 in groups C and D improved joint structural damage in rat OA, alleviated inflammatory reactions, downregulated MMP-13 expression, and increased collagen type Ⅱ expression.
CONCLUSION:VX765 can improve rat OA and reduce chondrocyte inflammation, possibly through the activation of the Nrf2 pathway.
