[Electroacupuncture down-regulates expression of ADAMTS-4 of intervertebral disc in rats with lumbar intervertebral disc degeneration].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation on the levels of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) protein in the lumbar intervertebral disc tissue and serum in prolapsed lumbar intervertebral disc degeneration (PLIDD) rats, so as to explore its mechanism underlying improvement of intervertebral disc degeneration (IDD). METHODS: A total of 48 Sprague-Dawley rats were divided into sham operation (n＝12), model (n＝18) and EA (n＝18) groups. The PLIDD model was established by puncturing the lumbar discs (L4-L5, L5-L6) with a gauge-22 syringe needle. After modeling, EA stimulation was applied to "Pangguangshu"(BL28), "Zusanli"(ST36) and "Zhijian"for 20 min, 6 times per week for 4 weeks. The lumbar intervertebral disc tissue and blood samples were collected at the 4th, 6th and 8th week after modeling, respectively. The expression of ADAMTS-4 in the lumbar intervertebral disc tissue was detected by immunohistochemistry and Western blot (WB), separately. The content of ADAMTS-4 in the serum was detected by ELISA. RESULTS: Both immunohistochemical stain and WB showed that the expression levels of lumbar ADAMTS-4 at the 4th, 6th and 8th week were significantly up-regulated in the model group relevant to the sham operation group (P<0.01). Following EA treatment, the expression levels of ADAMTS-4 on day 14 and 28 were notably lower in the EA group than in the model group (P<0.05). The serum ADAMTS-4 contents were significantly increased in the model group than in the sham operation group at the 3 time-points (P<0.05, P<0.01), and considerably decreased in the EA group than in the model group on day 28 after EA intervention(P<0.05)．. CONCLUSION: EA can down-regulate the ADAMTS-4 expression of lumbar intervertebral disc tissue in PLIDD rats, which may contribute to its effect in relieving lumbar intervertebral disc degeneration.

Luteolin suppresses inflammation through inhibiting cAMP-phosphodiesterases activity and expression of adhesion molecules in microvascular endothelial cells.

Luteolin, an anti-inflammatory ingredient found in the Chinese herb Folium perillae, can inhibit not only the cyclic adenosine monophosphate (cAMP)-phosphodiesterases (PDEs) activity of neutrophils, but also the expression of lymphocyte function-associated antigen-1 in neutrophils, both of which result in a decrease in the adhesion between neutrophils and microvascular endothelial cells. However, the effect of luteolin on the cAMP-PDEs activity and expression of adhesion molecules in endothelial cells are not clear. In the present study, primary rat pulmonary microvascular endothelial cells and a lipopolysaccharide-induced rat acute pneumonia model were used to explore the role of luteolin on cAMP-PDEs activity, expression of adhesion molecules, and leukocyte infiltration. We demonstrate that rat pulmonary microvascular endothelial cells expressed high levels of cAMP-PDEs, specifically PDE4, and further luteolin exhibited dose-dependent inhibition on the activity of cAMP-PDEs or PDE4 in endothelial cells. Luteolin also had a significant inhibitory effect on the expression of vascular cell adhesion molecule (VCAM)-1, but not intracellular cell adhesion molecule (ICAM)-1 in microvascular endothelial cells. Further, we show that luteolin decreased the levels of soluble ICAM-1 (sICAM-1), but not soluble E-selectin in the serum of rats subjected to acute pneumonia. We also show that luteolin treatment decreased the wet/dry weight ratio of lung tissue and reduced the total number of serum leukocytes in a dose-dependent manner in a rat acute pneumonia model. In conclusion, these results demonstrate that luteolin suppresses inflammation, at least in part, through inhibiting both cAMP-PDEs or PDE4 activity and the expression of VCAM-1 (in vitro) and sICAM-1 (in vivo) in endothelial cells.

Luteolin Partially Inhibits LFA-1 Expression in Neutrophils Through the ERK Pathway.

Luteolin inhibits the adhesion of neutrophils to microvascular endothelial cells and plays an important anti-inflammatory role, owing to its mechanism of suppressing the expression of lymphocyte function-associated antigen-1 (LFA-1) in the neutrophils. Our study deals with the different signaling pathways participating in LFA-1 expression in neutrophils along with the regulation of luteolin in order to elucidate new anti-inflammatory targets of luteolin, thus providing a basis for clinical applications. In our study, neutrophils were separated using density gradient centrifugation and the cAMP levels were determined using ELISA. Additionally, phosphorylation levels of p38 mitogen-activated protein kinase (MAPK), extracellular regulated protein kinase (ERK), phosphatidylinositol-3-kinase (PI3K), and Janus kinase (JAK) were also detected by Western blotting. LFA-1 expression was estimated using flow cytometry. The results showed that inhibiting agents used against p38 MAPK, ERK, PI3K, and JAK could significantly inhibit LFA-1 expression on neutrophils (p < 0.05, p < 0.01). Luteolin also induced a noteworthy elevation of cAMP in neutrophil supernatants (p < 0.01). It could also significantly inhibit ERK phosphorylation (p < 0.05, p < 0.01), and had no obvious effect on p38 MAPK phosphorylation in neutrophils (p > 0.05). However, phosphorylation of PI3K and JAK was not detected in neutrophils. To conclude, the p38 MAPK, ERK, PI3K, and JAK pathways are involved in the regulation of LFA-1 expression in neutrophils, and luteolin partially inhibits LFA-1 expression by increasing cAMP levels and suppressing ERK phosphorylation.

Photomanipulated Architecture and Patterning of Azopolymer Array.

Here reported is the approach to prepare the tunable 3D architecture and patterning through photoinduced orientation of azopolymer. The hemispherical PAzoMA array can be transformed into spindlelike, flat ellipsoidlike, thick spindlelike, near-hexagon, near-quadrangle, and near-rhombus arrays while being exposed to linearly polarized light (LPL). The size and alignment of the arrays can be precisely controlled by manipulating the irradiation time. Furthermore, complex 3D architectures of the PAzoMA array are readily fabricated through secondary irradiation along different direction. This technique is promising for functionalized surfaces and photonic devices.