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The Role of Angiopoietin-like Protein4in Severe Acute Pancreatitis-associated Lung Injury of Rats and Rat Pulmonary Microvascular Endothelial Cells

Author: WangYuZuo
Tutor: ChenHaiLong
School: Dalian Medical University
Course: Surgery
Keywords: angiopoitin-like protein4 pulmonary microvascular endothelial cellF-actin severe acute pancreatitis-associated lung injury rat
CLC: R657.51
Type: PhD thesis
Year: 2013
Downloads: 43
Quote: 0
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Abstract


Background: Severe acute pancreatitis associated lung injury is an inflammatorydisease which closely resembles the severe acute pancreatitis (SAP), whichcharacterized by tissue injury and a systemic inflammatory response, but themechanisms are unknown. Pulmonary microvascular endothelial cells (PMVECs)possess both highly proliferative and angiogenic capacities, and lined at the criticalinterface between the blood and microvessel are primary targets of inflammatorycytokines during lung inflammation. Angiopoietin-like4(Angptl4) is a circulatingprotein that has recently been implicated in the regulation of angiogenesis andmetastasis. This study aimed to investigate the effect of Angptl4on RPMVECs andsevere acute pancreatitis-associated lung injury of rats.Methods: SAP animal model was administrated as previously method andadministrated by Rosiglitazone (ROSI, ROZ) and GW9662. The cell culture wasadministrated by LPS (100ng/mL). The PCR product was recombined topcDNA3.1-eGFP vector and pcDNA3.1-eGFP-Angptl4vector, which were transfectedinto RPMVECs with SuperFect Transfection Reagent (QIAGEN). Vascular endothelialgrowth factor (VEGF) was detected from vascular endothelial cells of lung byimmunohistochemistry. Pancreatitis tissue was stained with hematoxylin-eosin andimmunohistochemistry analysis with BCL-2antibodies was performed.Angptl4mRNAlevels, protein level and cell morphology of RPMVECs in experiment group weredetected with RT-PCR, real time-PCR, Western-blotting, MTT, ELISA and Confocaldevelopment methods, respectively.Results: Angptl4expression vector pcDNA3.1-eGFP-Angptl4was successfullyconstructed. The Angptl4mRNA level in LPS-pcDNA3.1-eGFP was slightly increasedand experiment group was significantly higher than that of empty vector group and blank control group with obviously differences. The pro-apoptosis casapase8,9andBax protein was inhibited, while p-AKT/AKT and p-Mek1/2protein expression wasalso decreased. Angptl4protein was increased after induced by deoxycholic acidsodium salt, expansion with PPAR-γagonist rosiglitazone treatment, whereas, it wasobviously decreased with the PPAR-γantagonist GW9662(p<0.01). The rosiglitazonegroup had obviously decreased levels of the inflammatory cytokine TNF-α(p<0.01)and inhibit expression of VEGF in the vascular endothelial cells of lung associatedpancreatitis. Overexpression of Angptl4can inhibit that LPS cause the increase ofpermeability of RPMVEC, which relates to the depolymerization of central F-actin inRPMVECs.Conclusions: In summary, our study demonstrated that overexpression of Angptl4may induce protective, anti-inflammatory and antiangiogenic effectiveness. It representsa novel therapeutic target gene for the therapy of severe acute pancreatitis-associatedlung injury and ALI induced by LPS.

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CLC: > Medicine, health > Surgery > Of surgery > Abdominal surgery > Pancreas > Pancreatitis
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