Title:Astragaloside IV Protects Against IL-1β-Induced Chondrocyte Damage via Activating Autophagy
Volume: 24
Issue: 11
Author(s): Hang Xu, Wang Jing-Bo, Yue-Ping Chen*, Wei Huang and Zong-Bo Wei
Affiliation:
- Department of Orthopedics,
Ruikang Hospital Affiliated with Guangxi University of Chinese Medicine, Nanning, China
Keywords:
Astragaloside IV, autophagy, chondrocyte, osteoarthritis, inflammation, interleukin-1β.
Abstract:
Background: Osteoarthritis (OA) is a chronic inflammatory condition that
affects the articular cartilage. Astragaloside IV (AS-IV) constitutes the primary active
component of the Chinese herbal medicine Huangqi (Radix Astragali Mongolici). AS-IV
demonstrates anti-inflammatory and anti-apoptotic attributes, exhibiting therapeutic
potential across various inflammatory and apoptosis-related disorders. Nevertheless,
its pharmaceutical effects in OA are yet to be fully defined.
Objectives: This study aimed to investigate the protective impact of AS-IV on rat
chondrocytes treated with IL-1β and ascertain whether autophagy plays a role in this
effect.
Methods: Chondrocytes were isolated and cultivated from the knee joints of neonatal
SD mice. The study included the blank control group, the model group, and the AS-IV
concentration gradient group (50, 100, 200 μmol/L) to intervene with chondrocytes.
The MTT assay was employed to assess cell viability at varying culture periods,
enabling the determination of suitable concentration and duration. Subsequently,
chondrocytes were treated with the optimal AS-IV concentration and divided into three
groups: the model group replicated IL-1β-induced inflammatory chondrocyte injury, the
AS-IV group received a co-culture of AS-IV and IL-1β, and a blank control group was
established. Changes in cell morphology and structure were observed using ghost pen
cyclic peptide staining. ELISA was used to measure TNF-α and GAG levels in cell
supernatants. RT-qPCR assessed p62 and LC3 mRNA expression, while Western Blot
evaluated p62 and LC3Ⅱ/Ⅰ protein expression.
Results: AS-IV promoted chondrocyte proliferation and concurrently inhibited cell
apoptosis. An optimal AS-IV dose of 200 μmol/L and a suitable reaction time of 48 h
were identified. Ghost pen cyclic peptide staining indicated that the model group's
cytoskeleton exhibited fusiform changes with reduced immunofluorescence intensity,
as opposed to the blank control group. The AS-IV group displayed more polygonal
cytoskeletal morphology with increased immunofluorescence intensity. AS-IV reduced
TNF-α levels and elevated GAG levels in the culture supernatant. Additionally, AS-IV
lowered p62 mRNA and protein expression while increasing LC3 mRNA expression in
cultured chondrocytes.
Conclusion: Our findings suggest that AS-IV mitigates inflammatory chondrocyte
injury, safeguarding chondrocytes through a potential autophagy suppression
mechanism. These results imply that AS-IV could offer preventive advantages for OA.
