Bushen Qiangjin capsule inhibits the Wnt/α-catenin pathway to ameliorate papain-induced knee osteoarthritis in rat.

OBJECTIVE: To evaluate the molecular mechanism underlying the beneficial effect of Bushen Qiangjin capsule (BSQJ), a Traditional Chinese Medicine, on knee osteoarthritis (KOA). METHODS: In the present study, 32 female Sprague-Dawley rats were randomly divided into four groups: control, KOA, high-dose BSQJ (H-BSQJ), and low-dose BSQJ (L-BSQJ). After successfully establishing the KOA model by intra-articular injection of papain, H-BSQJ and L-BSQJ groups were intragastrically administered 0.243 and 0.122 g/kg BSQJ, respectively, daily for 6 weeks. At the end of the experiment, knee articular cartilage tissues of rats were collected for evaluation by hematoxylin and eosin staining, Safranin O-Fast Green staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay. Serum interleukin-1α and tumor necrosis factor-α levels of rats were detected with an enzyme-linked immunosorbent assay method. Gene expression of Wnt-4, α-catenin, Frizzled-2, glycogen synthase kinase-3ß (GSK-3ß), cysteinyl aspartate-specific proteinases 3 and 9 (caspases 3 and 9), collagen type II alpha 1 (Col2a1), and matrix metalloproteinases 1 and 13 (MMP-1 and MMP-3) of rat knee articular cartilage was quantified by reverse transcription-quantitative polymerase chain reaction analysis. Wnt-4, α-catenin, Frizzled-2, GSK-3ß, cleaved caspase-3, and cleaved caspase-9 protein expression in rat knee articular cartilage was determined by western blot analysis. RESULTS: BSQJ obviously reduced pathological damage and matrix degradation of articular cartilage in KOA rats. Compared with the KOA group, H-BSQJ rats exhibited downregulated mRNA and protein expression of Wnt-4, ß-catenin, Frizzled-2,and caspase-3, as well as upregulated mRNA and protein expression of GSK-3α. In addition, H-BSQJ significantly increased mRNA expression of Col2a1 and decreased mRNA expression of MMP-1 and MMP-13. CONCLUSION: BSQJ exerted a beneficial effect on KOA by a mechanism involving downregulation of the Wnt/α-catenin pathway, which inhibited both cartilage extracellular matrix degradation and chondrocyte apoptosis to ameliorate KOA in rats.

The synovial fluid neuropeptide PACAP may act as a protective factor during disease progression of primary knee osteoarthritis and is increased following hyaluronic acid injection.

The correlation of serum and synovial fluid (SF) pituitary adenylate cyclase-activating polypeptide (PACAP) levels with disease progression of primary knee osteoarthritis (OA) was explored. Radiographic severity of OA was determined by Kellgren-Lawrence (K-L) grades. PACAP levels were measured by ELISA before treatment, and 4 and 8 wk following hyaluronic acid (HA) injection. Levels of IL-1ß and MMP-3 were also detected. The numeric pain scale (NPS), revised Oxford Knee Score (OKS), and American Knee Society Score (AKSS) were employed to evaluate to symptomatic severity. Receiver-operating-characteristic (ROC) curve analysis was carried out to compare the diagnostic value of PACAP, IL-1ß, and MMP-3 for the K-L grade. PACAP concentrations in SF but not serum were significantly lower in OA patients compared with controls. SF PACAP levels were negatively associated with K-L grades and higher NPS as well as worse AKSS and OKS. Further analysis demonstrated that PACAP concentration in SF was negatively correlated with expressions of IL-1ß as well as MMP-3 and may act as a marker for radiographic progression along with MMP-3. Last, we found SF PACAP levels exhibited an incremental trend after HA injection. These findings confirmed the crucial role of PACAP deficiency in the development of primary knee OA.

[Influence of Jiangu decoction on peroxisome proliferator-activated receptor (PPAR) in the femoral head of rabbits with steroid-induced femoral head necrosis].

OBJECTIVE: The Jiangu decoction is used in the treatment of steroid-induced femoral head necrosis in clinical experiences, which has functions of tonifying kidney and activating blood, and invigorating spleen to remove phlegm. The decoction is mainly composed of Radix Polygoni Multiflori, Rhizoma alismatis Rhizoma Drynariae, haw, medlar, Radix Astragali, radix rehmanniae, angelica, Radix Codonopsis, radix salviae miltiorrhizae, Fructus Ligustri Lucidi, licorice, pharmaceutical composition. This study was designed to investigate the influence of Jiangu decoction on peroxisome proliferator-activated receptor gamma (PPARgamma) in the femoral head of rabbits with steroid-induced femoral head necrosis. METHODS: Eighteen adult SPF healthy New Zealand rabbits were divided into 3 groups: control group, model group, Jiangu decoction group. The rabbits of Jiangu decoction group orally received Jiangu decoction suspension with a dose of 10 ml/kg each day and the drug content was 0.719 g/ml. The rabbits in control and model groups were given saline with a dose of 10 ml/kg. The methylprednisolone sodium succinate was injected intramuscularly into left leg with a dose of 40 mg/kg. Then the rabbits were fed continuously for 3 weeks. The glucocorticoid levels, PPARgamma and plasma glucocorticoid levels in the femoral head were measured before and after modeling. RESULTS: Before model established, the plasma glucocorticoid levels had no significant difference among three groups (P=0.301). At 3 weeks after model established,the plasma glucocorticoid level of rabbits in model group increased compared to the control group (P=0.001); and the plasma glucocorticoid level of rabbits in Jiangu decoction group decreased compared with model group (P=0.001). The glucocorticoid level in the local femoral head of rabbits in model group increased compared to the control group (P=0.001); and the glucocorticoid level in the local femoral head of rabbits in Jiangu decoction group decreased compared with model group (P=0.001). The PPARgamma level in the local femoral head of rabbits in model group increased compared to the control group (P=0.018);and the PPARgamma level in the local femoral head of rabbits in Jiangu decoction group decreased compared with model group (P=0.033). CONCLUSION: The Jiangu decoction is effective to inhibit the femoral head adipogenic differentiation by decrease the PPAR content, so as to prevent and treat steroid-induced femoral head necrosis.