ABSTRACT
Objective To evaluate the effect and mechanism of rivaroxaban, an inhibitor of coagulation factor Ⅹa (FⅩa), on endotoxin-induced injury to human umbilical vein endothelial cells (HUVEC). Methods When cultured HUVEC grow to 80% fusion, they were divided into four groups according to the random number method: blank control group (DMEM medium), lipopolysaccharide (LPS) group (cells were challenged by 100 μg/L LPS for 16 hours), FⅩa+LPS group (cells were challenged by LPS for 16 hours after they were cultured with 100 nmol/L FⅩa for 24 hours), and FⅩa+RIV+LPS group (cells were challenged by LPS for 16 hours after they were cultured with 100 nmol/L FXa and 1 μmol/L rivaroxaban for 24 hours). After each group of cells were challenged with LPS, the cell activity was detected by the cell proliferation and toxicity kit (CCK-8); the cell migration ability was detected by cell scratch experiments;the abilities of cells migration were measured by scratch-wound-healing assay; the apoptosis of cells were evaluated using flow cytometry; the endothelial barrier of cells was assessed by Transwell and Evans blue; the levels of tumor necrosis factor-α(TNF-α), interleukin (IL-1β, IL-6) were detected by the enzyme linked immunosorbent assay (ELISA); the expressions of nuclear factor-κB (NF-κB) and mitogen activated protein kinase (MAPK) signaling pathway were detected by Western Blot. Results Compared with blank control group, the cell viability in LPS group was significantly decreased, and the migration ability, number of apoptotic cells, and barrier permeability of endothelial cells was significantly increased, the levels of TNF-α, IL-1β and IL-6 were significantly increased, and the expressions of phosphorylation of c-Jun N-terminal kinase (p-JNK), phosphorylation of p38MAPK (p-p38MAPK), phosphorylation of transforming growth factor kinase 1 (p-TAK1) and phosphorylation of NF-κBp65 (p-NF-κBp65) were significantly increased. It indicated that LPS could stimulate the inflammatory response of vascular endothelial cells, and had a significant impact on cell activity, apoptosis and function. There was no significant difference in above indexes between FⅩa+LPS group and LPS group, except for the level of IL-6 being higher in FⅩa+LPS group. Compared with FⅩa+LPS group, in FⅩa+RIV+LPS group, the cell activity was significantly increased (A value: 0.42±0.02 vs. 0.33±0.02), and migration ability was significantly decreased (folds: 1.78±0.17 vs. 2.24±0.20), the number of apoptotic cells was significantly decreased [(11.30±0.70)% vs. (21.03±0.19)%], and permeability of monolayers endothelial cells was significantly decreased [(149±12)% vs. (253±15)%], the levels of inflammatory cytokines were significantly decreased [IL-1β(ng/L): 163.2±20.7 vs. 477.8±20.2, IL-6 (ng/L): 69.3±0.5 vs. 238.0±24.1, TNF-α(ng/L): 117.0±13.1 vs. 196.2±4.5], the expressions of p-TAK1 and p-NF-κBp65 were significantly decreased (p-TAK1/TAK1: 0.74±0.09 vs. 1.85±0.15, p-NF-κBp65/NF-κBp65: 1.15±0.17 vs. 2.36±0.20), with statistically significant differences (all P <0.05). There was no significant difference in the p-JNK, p-p38MAPK expressions between FⅩa+RIV+LPS group and FⅩa+LPS group (p-JNK/JNK: 1.64±0.12 vs. 1.65±0.15, p-p38MAPK/p38MAPK: 2.31±0.32 vs. 2.35±0.20, both P > 0.05). Conclusion Rivaroxaban can effectively relieve the inflammatory response of HUVEC stimulated by LPS, which may be related to the inhibition of NF-κB signaling pathway activation rather than MAPK signaling pathway.
ABSTRACT
Objective The influence of myasthenia gravias(MG) upon the long term survival of thoma patients varied in the retrospective studies as reported,including the positive,negative impact and irrelevant influence.The recent study from the Chinese Alliance for Research on Thynic Diseses(ChART) indicated that in different stages of thymoma,MG influenced the prognosis in different ways.It's the author's conclusion that the complicated influences of MG on the prognosis of thymoma include a direct one which is negative resulting from MG related complications and an indirect positive one due to its beneficial pathologic and staging patterns.Inprovement of the prognosis for patients with thymoma complicating MG will be expected with advancement in MG therapeutics.
ABSTRACT
Objective Portal hypertension and ischemia/reperfusion (I/R) have been implicated in small-for-size liver graft dysfunction. Matrix metalloproteinases-2 (MMP-2) and MMP-9 are critically involved in hepatic I/R injury. The goal of this study was to investigate the role of MMP-2 and MMP-9 in acute small-for-size graft injury. Methods 108 rats were divided into three groups:100 % (full-size), 50 % (half-size) and 25 % (quarter-size) liver transplantation groups. Blood and liver samples were collected to assess liver function, hepatic malondialdehyde (MDA) content, tissue myeloperoxidase (MPO) activity and histological changes. ELISA, real-time PCR, gelatin zymography, and immunohistochemistry were used to determine the expression pattern of MMP-2 and MMP-9 in liver grafts. Results The expression levels of MMP-9 were significantly higher in quarter-size and half-size grafts than those in full-size liver grafts 6, 12, and 24 h after reperfusioa And theelevated levels of MMP-9 were related to graft size inversely. However, MMP-2 was expressed and remained in all groups invariably. MMP-9 overexpression was accompanied by extensive liver I/R injury, as evidenced by significant increases in hepatic microscopic damage scores, MDA content,MPO activity and liver function levels. Furthermore, MMP-9 was found mainly to locate around periportal area. The presence of the active form of MMP-9 was significantly higher in small-for-size grafts, which was correlated with sinusoidal dilatation, congestion and hemorrhage. Conclusion These results support critical function of MMP-9 in acute small-for-size liver graft injury. Moreover,portal hypertension may be a crucial trigger for expression and activation of MMP-9.