ABSTRACT
Objective:To investigate the protective effect of overexpressed tripartite motif containing (TRIM27) on severe acute pancreatitis (SAP) in mice and its possible mechanism.Methods:Twenty-four mice were randomly divided into the sham operation + control virus group (AAV-GFP group), sham operation + overexpression of TRIM27 group (AAV-TRIM27 group), SAP + control virus group (SAP+AAV-GFP group), SAP + overexpression of TRIM27 group (SAP + AAV-TRIM27 group), with 6 mice in each group. SAP model of mice was established by intraperitoneal injection of L-arginine (4 mg/kg). The sham operation group was injected with equal volume of normal saline, and the virus group was injected with control or TRIM27 overexpression adeno-associated virus (2×10 11 μg/ per mice). The serum and pancreatic tissue samples were collected 72 h after modeling. The levels of serum amylase, lipase, tumor necrosis factor α (TNF-α), interleukin-1b (IL-1b), IL-6, macrophage chemoattractant protein-1 (MCP-1) and the expression of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) in pancreatic tissue were detected by enzyme-linked immunosorbent assay. Hematoxylin eosin staining was used to observe the pathological damage of pancreatic tissue. The expressions of myeloperoxidase (MPO) and Ly6g positive inflammatory cells in mouse pancreas were observed by immunohistochemistry. The expression of p-p65, p65, p-ASK1, ASK1, p-JNK, JNK, p-p38 and p38 in pancreatic tissue were detected by Western blot. Results:The expression of TRIM27 in pancreatic of mice was significantly down regulated after SAP ( P<0.05); after overexpression of TRIM27 by adeno-associated virus, the expression of TRIM27 in mouse pancreas was significantly up-regulated ( P<0.05). There was no significant difference in the indexes of mice between the AAV-GFP group and AAV-TRIM27 group ( P>0.05). Compared with the SAP + AAV-GFP group, the levels of serum amylase, lipase, TNF-α, IL-1b, IL-6 and MCP-1 in mice of the SAP + AAV-TRIM27 group were significantly decreased, MDA in pancreatic tissue was decreased, SOD and GSH were increased, MPO and Ly6g inflammatory cells were significantly decreased, and p-p65, p-ASK1, p-JNK, and p-p38 protein expression were down regulated. Conclusions:Overexpression of TRIM27 alleviates SAP in mice by inhibiting inflammatory response and oxidative stress, and its mechanism may be through inhibiting NFκB/MAPK signaling pathway.
ABSTRACT
AIM:To investigate the expression characteristics of TRIM 27 in human nasopharyngeal carcinoma tissues, nasopharyngeal carcinoma 5-8F cells and NP69 cells, and to observe the effect of TRIM27 on the proliferation, in-vasion and migration of 5-8F cells.METHODS:The levels of TRIM27 in the nasopharyngeal carcinoma tissues and normal nasopharyngeal epithelial tissues were observed by the method of immunohistochemistry .The mRNA and protein levels of TRIM27 in the 5-8F cells and NP69 cells were determined by real-time PCR and Western blot .TRIM27 siRNA was trans-fected into the 5-8F cells with Lipofectamine 2000.The relative mRNA expression of TRIM27 was detected by real-time PCR.The relative protein expression of TRIM 27 was detected by Western blot .The cell proliferation was analyzed by CCK-8 assay and cell colony formation assay .The change of cell invasion was examined by Matrigel invasion assay .The change of cell migration were examined by wound healing assay .RESULTS:The results of immunohistochemistry showed that the protein expression of TRIM27 in the nasopharyngeal carcinoma tissues was obviously higher than that in the normal nasopha -ryngeal epithelial tissues .The results of real-time PCR and Western blot showed that the mRNA and protein levels of TRIM27 in the 5-8F cells were obviously higher than those in the NP69 cells.The abilities of proliferation, invasion and migration in the 5-8F cells were significantly suppressed after TRIM27 gene silencing ( P <0.05).CONCLUSION:TRIM27 acts as a oncogene in the 5-8F nasopharygeal carcinoma cells .The abilities of proliferation , invasion and migration are significantly suppressed after TRIM27 gene silencing in the 5-8F cells.