Identification and characteristics of microRNAs in rat abdominal aortic aneurysms
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ABSTRACT: Abdominal aortic aneurysm (AAA) is a lethal disease, occurring mostly in men more than 65 years of age. Until recently, the pathogenesis of AAA remains poorly understood. MicroRNAs (miRNAs) are a novel class of endogenous small noncoding RNAs that play important roles in diverse biological and pathological processes and was more recently investigated in cardiovascular physiology and pathology. In this study, we employed microarray to detect and compare miRNA expressions of AAAs in rats. Four miRNAs were validated using real time RT-PCR. Functional annotations of putative targets of deregulated miRNAs via bioinformatics approaches revealed that predicted targets were highly enriched and involved in several signaling pathways important for AAA formation. Our results indicate that miRNAs are extensively involved in rat AAA formation and provide a global view of AAA miRNA profiles, which is expected to provide new clues to develop targeted therapies against this calamitous disease. Sprague-Dawley rat AAA model was established by calcium chloride and collagenase co-incubation method. After 28 days, three rats with confirmed AAAs and three normal rats form control(sham operation) group were euthanized and the aortas specimens were used for this experiment.
Project description:In this study we used microarrays to examine relative genes expression within the aorta of ApoE-/- infused with angiotensin II in relation to aneurysm formation. Infusion of angiotensin II induces aortic dilatation particularly of the suprarenal aorta in ApoE-/- mice. Based on studies carried out in our and other laboratories the response to angiotensin II is variable, with some mice developing large aneurysms but other animals appearing resistant to aneurysm formation with aortic diameters similar to that of saline controls. We compared RNA expression from whole aortas of 17 week old male ApoE-/- mice exposed to angiotensin II (1.44 µg/kg/min) for 4 weeks where there was clear evidence of aortic aneurysm formation (n=5) with that of mice failing to develop aneurysms (n=7) and those exposed to saline infusion (n=6). AAA was defined as diameter of suprarenal aorta greated than 1.5mm measured on photographs of aortas at necroscopy. Keywords: Disease state analysis 18 samples analysed, AAA (n=5), no AAA (n=7), saline (n=6). AAA - abdominal aortic aneurysm
Project description:Abdominal aortic aneurysm (AAA) is a lethal disease, occurring mostly in men more than 65 years of age. Until recently, the pathogenesis of AAA remains poorly understood. MicroRNAs (miRNAs) are a novel class of endogenous small noncoding RNAs that play important roles in diverse biological and pathological processes and was more recently investigated in cardiovascular physiology and pathology. In this study, we employed microarray to detect and compare miRNA expressions of AAAs in rats. Four miRNAs were validated using real time RT-PCR. Functional annotations of putative targets of deregulated miRNAs via bioinformatics approaches revealed that predicted targets were highly enriched and involved in several signaling pathways important for AAA formation. Our results indicate that miRNAs are extensively involved in rat AAA formation and provide a global view of AAA miRNA profiles, which is expected to provide new clues to develop targeted therapies against this calamitous disease.
Project description:We performed two different pools for the 10 AAA patients (n=5 AAA patients in pool A and n=5 AAA patients in pool B) and two different pools for the 10 healthy subjects (n=5 controls in pool C and n=5 controls in pool D). Two replicates of the two microarray experiments were made. Experiment 1: AAA pool A Cy3 labeled vs control pool C Cy5 labeled AAA pool A Cy5 labeled vs control pool C Cy3 labeled (dye swap); Experiment 2: AAA pool B Cy3 labeled vs control pool D Cy5 labeled AAA pool B Cy5 labeled vs control pool D Cy3 labeled (dye swap).
Project description:We sought to identify differentially regulated microRNAs in infrarenal mouse aortic tissue after AAA-induction with PPE, compared with sham-operated mice. This treatment leads to rapid development of infrarenal aortic aneurysms with significant diameter differences observed by Day 7. We found 41 miRNAs were up-regulated with aneurysm and 37 down-regulated at p<0.05, which were also altered by >1.5-fold. Utilizing the PPE infusion model, we induced AAA in Male 10-week-old C57/Bl6 mice, 7 days after AAA-induction with PPE. One array per mouse, 5 mice per group, two groups (PPE and sham).
Project description:Abstract: The pathogenesis of AAA involves vascular inflammation and oxidative stress. Astragali Radix contains cycloastragenol (CAG) known to have anti-inflammatory and anti-oxidative properties. We hypothesized that CAG supplement impairs AAA progression. AAA was induced in male rats by intraluminal elastase infusion in the infrarenal aorta and treated daily with CAG (125 mg/kg/day). Aortic expansion was followed weekly by ultrasound, with euthanization at day 28. Changes in AAA wall composition were analyzed at mRNA levels, histology, zymography and explorative proteomic analyses. At day 28, mean AAA diameter was 37% lower in CAG group (p<0.0001). In aneurysm cross sections, elastin content was insignificantly higher in CAG group (10.5% ± 5.9% vs 19.9% ± 16.8%, p=0.20) with more preserved elastin lamellae structures (p=0.0003) and with no microcalcifications. Aneurysmal matrix metalloprotease-2 activity was reduced by CAG treatment (p=0.022), and mRNA levels of inflammatory- and antioxidative markers showed no difference between groups. Explorative proteomic analysis showed no difference in protein levels when adjustment for multiple testing. Amongst unadjusted affected proteins were fibulin-5 (p=0.02), aquaporin-1 (p=0.02) and prostacyclin synthase (p=0.006). CAG impairs experimental AAA progression by reduction of elastin degradation through decreased MMP-2 activity, thus CAG could be considered tested in AAA patients.
Project description:To understand the molecular change occurring in the atria during obstructive sleep apnea, we have implemented a rat model of OSA involving the surgical implantation of a tracheal obstructive device, allowing the rats to remain conscious and free-roaming throughout 2 weeks of apnea administration. Rats were divided into severe and moderate apnea groups, receiving a 23 second or 13 second apneas per minute, respectively. The two-dimensional polyacrylamide gel electrophoresis (2D PAGE) of atrial homogenates to compare the dysregulations in the protein pattern in severe and moderate apnea when compared to control.
Project description:We performed a global analysis of extracellular miRNAs in the serum of human subjects diagnosed with Alcohol Use Disorder (AUD) to identify robust biomarkers of early brain damage or dysfunction. This was performed in a set of 20 AUD subjects and 10 age-matched controls. They were subjected to comprehensive medical, neuropsychological and neuroimaging tests, followed by comparison of miRNA levels found in peripheral blood serum. The levels of miRNAs were quantified using two independent high-throughput methods: Affymetrix microarray and Illumina next-generation RNA-sequencing. Cross-species validation was performed using rat drinking models and mouse neural stem cell culture models, with tissue specificity of the serum miRNAs examined using additional RNA-sequencing of serum and body tissues in untreated rats. This data series includes the rat miRNA 2.0 GeneChip data only but is part of a larger SuperSeries (GSE71579). A total of 38 miRNA 2.0 GeneChips were run on small RNA purified from the rat blood serum samples.
Project description:We measured gene expression in the adrenal glands of the Spontaneously Hypertensive Rat (SHR) and Wistar-Kyoto rat (WKY) using Affymetrix RG-U34A GeneChips. All rats were aged-matched at 4-weeks. The rats were obtained from the colonies at the Univeristy of California San Diego, La Jolla, CA.
Project description:We applied Solexa sequencing technology to identify rat microRNA genes in dorsal root ganglia (DRGs) following sciatic nerve resection. Using Solexa sequencing, computational analysis and Q-PCR verification, 114 novel miRNAs in rats were discovered and identified, of which 52 novel miRNAs were first reported in rat DRGs and 62 novel miRNAs were produced at days 1, 4, 7 and 14 after sciatic nerve resection. These data provide an important resource relating to the role and regulation of miRNAs for future studies relating to peripheral nerve injury and regeneration. 18-30 nt small RNAs from 30 Thirty Sprague-Dawley (SD) rats were sequenced at one Solexa lane
Project description:Objective: This study aims to establish a T2DM rat model consistent with the natural history of the disease, and apply TMT proteomics technology to analyze the retina to reveal the pathogenic mechanism of NPDR and search for new targets for NPDR intervention. Methods: Six-week-old SD male rats were randomly divided into type 2 diabetes group (T2DM group) and normal group (NOR group). T2DM group rats were fed with high-fat diet containing 60% fat energy, while NOR group rats were fed with normal chow diet. After 6 weeks, oral glucose tolerance tests were conducted on the two groups of rats. Following confirmation of insulin resistance, the T2DM group rats were intraperitoneally injected with 2% STZ (30mg/kg), and blood glucose levels were monitored 72 hours later. The rats with random blood glucose levels higher than 16.7 mmol/L were fed with the high-fat diet for another 6 weeks, and then retinas were collected from the two groups of rats for TMT proteomic analysis. Results: After 72 hours and 6 weeks after STZ injection, the T2DM group rats showed typical symptoms of diabetes such as hyperglycemia, weight loss, increased food intake and water consumption, suggesting the establishment of a rat model of T2DM. The bioinformatics analysis results of proteomics reveal the close relationship between differentially expressed proteins, fatty acid metabolism, and angiogenesis. Conclusion: In a T2DM rat model consistent with the natural history of the disease, this study used TMT proteomics technology to deeply analyze the molecular mechanism of NPDR and revealed the key role of fatty acid metabolism in the pathogenesis of NPDR. In addition, Fabp3, Tinagl1, Col4a3, and Snrpd1, may subserve candidate targets for NPDR intervention.