ABSTRACT
This study is to investigate the effect of osthol on osteoclasts' activity, bone resorption as well as apoptosis in vitro, and explore the mechanism of osthol in preventing osteoporosis. Osteoclasts were separated from long-limb bones of new born rabbits, cultured in 24-well plate with glass slices and bone slices, and treated by 1 x 10(-5) mol x L(-1) osthol. Osteoclasts were identified by observing live cells with phase contrast microscope, HE staining, TRAP staining and toluidine blue staining of bone resorption pits. The numbers of bone resorption pits were counted as well as the surface area of bone resorption on bone slice. Osteoclasts were stained with acridine orange to detect the cell apoptosis. The ratio of apoptotic osteoclasts was observed under fluorescence microscope. The gene expression of RANKL, OPG, TRAP and p-JNK1/2 protein expression were examined using real time PCR and Western blotting, respectively. Comparing with the control group without osthol, the rates of apoptotic osteoclasts increased obviously and the number and area of bone resorption pits decreased evidently with 1 x 10(-5) mol x L(-1) osthol. There is significant difference between control group and experiment group treated by 1 x 10(-5) mol x L(-1) osthol. Therefore, the osthol through RANK+RANKL/TRAF6/Mkk/JNK signal pathway inhibits the osteoclasts activity, enhances osteoclasts apoptotic and inhibits the bone resorption.
Subject(s)
Animals , Rabbits , Acid Phosphatase , Metabolism , Apoptosis , Bone Resorption , Cells, Cultured , Cnidium , Chemistry , Coumarins , Pharmacology , Gene Expression , Isoenzymes , Metabolism , Mitogen-Activated Protein Kinase 8 , Metabolism , Mitogen-Activated Protein Kinase 9 , Metabolism , Osteoclasts , Metabolism , Pathology , Osteoprotegerin , Metabolism , Phosphorylation , Plants, Medicinal , Chemistry , RANK Ligand , Metabolism , Seeds , Chemistry , Signal Transduction , Tartrate-Resistant Acid PhosphataseABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect on early treatment with vacuum-assisted closure(VAC) to wound healing of acute explosion injury in pigs, and provide a new way for early treatment of battle wounds.</p><p><b>METHODS</b>Eight healthy 3-month Landrace pigs of both sexes with the body mass of (50 +/- 5) kg were selected in the study. Sixteen battle wounds were made by explosion of same type detonator (pattern number: 660929F48840-55, included DDNP 0.3 g, RDX 0.7 g) in hibateral skin of buttock of 8 pigs, which were divided into experimental group and control group (pair wounds of left and right). The raw sufaces were thorough debrided at 3 h after exposure, according to the characteristics of treatment on the battlefield, experimental group was treated with VAC under the pressure of (-50 +/- 5) Kpa after debridement and sterilization and control group was treated with routine dry sterile gauze draping. Results of bacteriology (bacterial counts and the proportion of G+ bacteria) and pathology (HE stain and Masson stain) were detected at every wound before and after treatment.</p><p><b>RESULTS</b>At the 3 days after treatment,the bacterial number in the experimental group was [(7.82 +/- 0.55) x 10(4) ] CFU/g, in control group was [(1.07 +/- 0.14) x 10(6)] CFU/g. There was significant difference between two groups. The proportion of G+ bacteria in experimental group was significantly increased. The raw surface in experimental group was clean with affluent and neoformative granulation tissue, blood vessels and collagen, necrotic tissue decreased obviously by pathological observation.</p><p><b>CONCLUSION</b>VAC could reduce the quantity of bacteria, improve the proportion of G+ bacteria, and promote the formation of granulation tissue and the healing of wound. The VAC for the treatment of battle wounds has a positive effect.</p>