ABSTRACT
Objective To evaluate the relationship between Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) signaling pathway and propofol-induced inhibition of endotoxin-induced release of tumor necrosis factor-alpha (TNF-α) from alveolar macrophages (AMs) of rats.Methods AMs extracted from adult male Sprague-Dawley rats were cultured and inoculated in 6-well plates (1 × 106 cells/well)and in 96-well plates (1×104 cells/well).The cells were divided into 5 groups (n=18 each) using a random number table:control group (group C),dimethyl sulfoxide group (group D),lipopolysaccharide (LPS) group (group L),propofol group (group P) and LPS plus propofol group (group L+P).The cells were continuously cultured with phosphate buffer solution in group C.Dimethyl sulfoxide was added at the final concentration of 5 mg/ml in group D.LPS was added at the final concentration of 1 μg/ml in group L.Propofol was added at the final concentration of 25 μmol/L (4.46 μg/ml) in group P.LPS and propofol were added at the final concentration of 1 μg/ml and 25 μmol/L (4.46 μg/ml),respectively,in group L+P.At 24 h of culture or incubation,the cell viability was detected by CCK-8 assay,the morphological changes of cells were observed using Wright's staining,the concentration of TNF-α in the supernatant was determined by enzyme-linked immunosorbent assay,and TLR4 expression and NF-κB activities were measured by Western blot.Results Compared with group C,the cell viability and concentration of TNF-α in the supernatant were significantly increased,the expression of TLR4 was up-regulated,and the activity of NF-κB was enhanced in L and L+P groups (P＜0.05),and no significant change was found in the parameters mentioned above in D and P groups (P＞0.05).Compared with group L,the cell viability and concentration of TNF-α in the supernatant were significantly decreased,the expression of TLR4 was down-regulated,and the activity of NF-κB was weakened (P＜0.05),the morphological changes of cells were significantly attenuated,and the number of pseudopodia was reduced in group L+P.Conclusion The mechanism by which propofol inhibits endotoxin-induced release of TNF-α from AMs is related to inhibited activation of TLR4/NF-λB signaling pathway in rats.
ABSTRACT
In order to enhance the anticoagulant properties of decellularized biological materials as scaffolds for tissue engineering research via heparinized process, the decellularized porcine liver scaffolds were respectively immobilized with heparin through layer-by-layer self-assembly technique (LBL), multi-point attachment (MPA) or end-point attachment (EPA). The effects of heparinization and anticoagulant ability were tested. The results showed that the three different scaffolds had different contents of heparin. All the three kinds of heparinized scaffolds gained better performance of anticoagulant than that of the control scaffold. The thrombin time (TT), prothrombin time (PT) and activated partial thromboplastin time (APTT) of EPA scaffold group were longest in all the groups, and all the three times exceeded the measurement limit of the instrument. In addition, EPA scaffolds group showed the shortest prepared time, the slowest speed for heparin release and the longest recalcification time among all the groups. The decellularized biological materials for tissue engineering acquire the best effect of anticoagulant ability in vitro via EPA heparinized technique.
Subject(s)
Animals , Anticoagulants , Chemistry , Biocompatible Materials , Chemistry , Heparin , Chemistry , Liver , Swine , Tissue Engineering , Tissue ScaffoldsABSTRACT
Objective To investigate the proliferation rule after autografl vein in autografl of vein and repairing arterial defect,discuss the minuteness structure changes related to time,and provide morphological date for the prevention and treatment of restenosis in clinic application.Methods Sixty canine femoral veins (3.5 cm and 5.5 cm) were excised,reversed and grafted to repairing femoral arterial defect using end- to-end anastomosis.During the period of 30-180 days excised the grafted veins,observed ultrastrural changes of endothelium,microstructural components relative content and patency in grafted veins through angiography, lipht microscope.Results Angiography show that the grafted vein wall get gradually thicker,vascular cavity get slender,and vessel elasticity shrinkage.Light microscope chalrify the ultrastructural changes of endotheh- um,the three lamine structure of grafted vascular wall could not be identified.Conclusion Grafted femoral veins had obviously arteriosclerosis trend after it was transplanted to the arterial system.Two months after the transplantation is the crisis time of the proliferation of the vascular wall.The effect of the proliferation of the grafted veins vascular wall and the impact of the hi-pressure of blood can lead some canine ruptured to die from pseudoaneurysms.