ABSTRACT
Objective:To investigate the possible mechanism of Wenjing Tongluo decoction （WTD） in alleviating articular cartilage defect in knee osteoarthritis （KOA） and delaying joint degeneration. Method:The KOA model was established by anterior cruciate ligament transection （ACLT）. Mice were classified into sham-operated group， model group， WTD high-dose and low-dose groups， and positive control group. Four weeks after modeling， WTD groups and the positive control group were given WTD （80， 20 g·kg<sup>-1</sup>） and glucosamine sulfate capsules （0.29 g·kg<sup>-1</sup>）， respectively， and the sham-operated group and model group received normal saline of the equivalent volume. After continuous intervention for 4 weeks， hemoxylin-eosin （HE） staining was used to observe the morphological changes of cartilage and Mankin scoring system was employed to score the knee cartilage. Western blot was combined with Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） to detect the protein and mRNA levels of vascular endothelial growth factor <italic>α</italic> （VEGFA）， vascular endothelial growth factor receptor 2 （VEGFR2）， extracellular signal-related kinase 1/2 （ERK1/2） and a disintegrin and metalloproteinase with thrombospondin motifs 4 （ADAMTS4）. Result:The Mankin score in the model group increased as compared with that in the sham-operated group （<italic>P</italic><0.01）. Compared with the model group， administration groups demonstrated alleviated articular cartilage defect and low Mankin score （<italic>P</italic><0.01）， but there was no statistical significance in Mankin score between the WTD groups and positive control group. The protein and mRNA levels of VEGFA， VEGFR2， ERK1/2， and ADAMTS4 in the model group were significantly higher than those in the sham-operated group （<italic>P</italic><0.01）. The protein expression of VEGFA and ERK1/2 was inhibited in each administration group as compared with that in the model group （<italic>P</italic><0.01）， and the inhibition in the positive control group was stronger than that in the WTD low-dose group （<italic>P</italic><0.05） but weaker than that in the WTD high-dose group （<italic>P</italic><0.01）. Glucosamine Sulfate capsules suppressed the expression of VEGFR2 and ADAMTS4 to the extent the same with low-dose WTD but weaker than the high-dose WTD （<italic>P</italic><0.05）. Conclusion:WTD can relieve the articular cartilage injury in KOA mice， and the mechanism may be related to VEGF/VEGFR2/ERK1/2 signaling pathway.
ABSTRACT
OBJECTIVE: To improve the stability and oral bioavailability of paeoniflorin, develop solid and hollow zein-paeoniflorin nanoparticles, and compare their properties and in vitro release. METHODS: The solid and hollow zein-paeoniflorin nanoparticles were fabricated by phase separation method, and the processing parameters were optimized according to the entrapment efficiency by orthogonal design. The morphology and mean size of nanoparticles were investigated and in vitro release was carried out using the optimal prescription. RESULTS: The mass of paeoniflorin, the concentration of zein and the stirring time in optimal prescription of solid zein-paeoniflorin nanoparticles were 10 mg, 10 mg•mL-1 and 2 h, respectively. These three factors in optimal prescription of hollow zein-paeoniflorin nanoparticles were 20 mg, 5 mg•mL-1 and 1 h, respectively. The obtained nanoparticles of solid and hollow zein-paeoniflorin nanoparticles were spherical with a mean particle size of 110 and 50 nm. And the drug load efficiency and entrapment efficiency of solid and hollow zein-paeoniflorin nanoparticles were 26.94%, 53.87%, 44.52% and 55.65%, respectively. In vitro release of both formulations indicated sustained release effect. CONCLUSION: The solid and hollow zein-paeoniflorin nanoparticles are successfully prepared by phase separation method. The method is simple and reliable. Compared with solid nanoparticles, the drug load efficiency of hollow nanoparticles is greatly improved, and in vitro studies of hollow zein-paeoniflorin nanoparticles show much slower release.