Inhibitory effects of berberine on osteoclast differentiation

Abstract

intervention followed by 3 to 7 days of washout period. Acupuncture in all groups was implemented at left LI10 with manipulation after insertion, 5 min, 10 min after insertion followed by needle removal. Microcirculation by laser Doppler perfusion imaging (LDPI) was measured 5 minutes before acupuncture till 10 minutes after removal of acupuncture. A repeated measures ANOVA was used for statistical analysis.Results: In each group, microperfusion increased by needle insertion was maintained till after needle removal, and different needle size induced different blood perfusion changes, with the increase in ST group significantly higher than SP, DT and DP groups (repeated measures ANOVA, P < 0.0001), but there was no significant difference by needling depth.Conclusion: ST intervention induced the greatest blood perfusion changes compared with other interventions, but the impact of different needling depths was not clear with blood perfusion changes. Studies investigating other physiological changes than blood perfusion by different acupuncture needle sizes and needling depths are needed.Contact: Xiuyu Zhang, Purpose: Indirubin-3'-monoxime (I3O), an active ingredient of Danggui Longhui Wan, exhibit antithrombotic activity through antiplatelet activity. Hence, we investigated the antiplatelet activity of I3O and the underlying mechanisms, focusing on the possible involvement of phospholipase C␥2 (PLC␥2) and extracellular signal-regulated kinase (ERK) 1/2. Methods: To identify the antithrombotic activity of I3O, we investigated using FeCl3-induced thrombus formation model, and platelet aggregation and coagulation ex vivo. In addition, the mechanism by which I3O is mediated the antithrombotic activity was assessed by determining platelet aggregation, immunoblotting, adenylyl cyclase activity, arachidonic acid (AA) liberation, and AA-metabolites conversion.Results: In a rat carotid artery injury model, oral administration (20 mg/kg/day) of I3O for 3 days significantly prolonged occlusion time, and ADP-and collagen-induced platelet aggregation, comparable with aspirin. In washed platelets in vitro, I3O potently inhibited collagen-induced platelet aggregation by suppression of PLC␥2 phosphorylation, consistent with the blockade of diacylglycerol (DAG) and AA formation, P-selectin secretion and the production of thromboxane B2 (TXB2). As expected, platelet aggregation induced by phorbol-12myristate 13-acetate (PMA), a protein kinase C (PKC) activator, was inhibited by I3O. Both I3O and U0126 markedly reduced collagen-induced phosphorylation of ERK1/2 and p47. Besides, I3O generally suppressed phosphorylation of JNK, p38, GSK-3␤, and AKT. Hence, we investigated to identify the effect of I3O in collagen receptor level; as a result, I3O concentrationdependently showed the inhibition pattern in immunofluorescence assay of glycoprotein VI (GPVI), as a collagen receptor. Moreover, I3O not only inhibited the phosphorylation of the tyrosine kinase Syk of GPVI but also suppressed the phosphorylation of PLC␥2 and ERK1/2 stimulated by...