Local and systemic alterations in signal transducers and activators of transcription (STAT) associated with human abdominal aortic aneurysms.
Local and systemic alterations in signal transducers and activators of transcription (STAT) associated with human abdominal aortic aneurysms.
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DOI：
10.1016/j.jss.2011.05.041
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发表时间：
2012-07
期刊：
影响因子：
--
通讯作者：
Eagleton MJ
中科院分区：
文献类型：
--
作者：
Liao M;Xu J;Clair AJ;Ehrman B;Graham LM;Eagleton MJ
信号转导和转录激活因子（STAT）蛋白是转录因子，当被磷酸化激活时，调节基因表达和细胞活性。本研究的目的是评估与腹主动脉瘤（AAA）相关的STAT蛋白的局部和全身表达和激活。在从经历AAA修复的患者（N=9）和从非动脉瘤（NA）供体（N=17）获得的主动脉壁样品中评估STAT蛋白的表达和活化。使用RT-PCR和免疫印迹（WB）测定评价主动脉样本中STAT 1、2、3、4、5a和5 b的mRNA和蛋白表达，并将其标准化为β-肌动蛋白（以任意单位表示）。使用磷酸化（p）-STAT特异性抗体，用WB测定法评估STAT活化。通过将p-STAT蛋白标准化为相应的总STAT水平来计算STAT活化的变化。通过评估来自AAA（AAA，N=8）、修复的动脉瘤（RA，N =8）或年龄/性别匹配的无AAA的对照（CT，N= 8）的循环白细胞中p-STAT的存在来评估STAT活化的全身性改变。进行流式细胞术以评估单核细胞、粒细胞和淋巴细胞中STAT 1（pY 701）、STAT 3（pY 705）和STAT 5a（pY 694）的循环水平。在基线和响应于用IFN-γ（50 ng/mL）或IL-6（100 ng/mL）的体外刺激进行评估。使用学生T检验分析结果，并表示为平均值±SEM。与NA相比，AAA组织中STAT-1（1.08±0.09 v.0.62 ±0.07）、-2（0.98±0.07 v.0.55 ±0.08）和-4（0.89±0.12 v.0.35 ±0.11）mRNA水平升高（均P<0.01）。STAT蛋白表达仅在STAT 1中相应增加（2.77±0.93 v.0.93 ±0.08，P<0.05）。与NA相比，AAA中p-STAT 2（0.77±0.1 v.0.1 ±0.02，P<0.01）、p-STAT 3（1.6±0.3 v.0.2 ±0.06，P<0.02）和p-STAT 5（0.57±0.03 v.0.2 ±0.03，P<0.05）水平的激活增加。在侵袭AAA外膜的炎性细胞中观察到磷酸化的STAT 1、2、3和5。此外，在AAA和NA中均观察到STAT 3的表达，而pSTAT 3仅在AAA中表达。p-STAT阳性循环白细胞的基线水平无差异。IFN-γ刺激使STAT-5a（pY 694）阳性CT淋巴细胞减少至基线的40±13%，但对AAA或RA淋巴细胞无影响（分别为116± 35%，102± 19%; P=0.01）。与AAA或RA粒细胞相比，STAT-5a（pY 694）阳性CT粒细胞也降低至基线的62±18%（分别为122± 25%，126± 17%; P=0.01）。在细胞因子刺激后，在白细胞中未观察到STAT 1（pY 701）和STAT 3（pY 705）的改变。STAT蛋白是转录活性的重要调节因子，并与心血管疾病有关。目前的数据表明，磷酸化STAT水平的改变与AAA相关。了解它们的作用可以进一步了解AAA形成的机制，并允许开发药物治疗方案。
Signal transducers and activators of transcription (STAT) proteins are transcription factors that when activated, by phosphorylation, regulate gene expression and cellular activity. The aim of this study was to evaluate the local and systemic expression and activation of STAT proteins associated with abdominal aortic aneurysms (AAA). Expression and activation of STAT proteins were assessed in aortic wall samples obtained from patients undergoing repair of AAA (N=9) and from non-aneurysmal (NA) donors (N=17). Aortic samples were evaluated for mRNA and protein expression for STAT1, 2, 3, 4, 5a, and 5b using RT-PCR and immunoblot (WB) assays and normalized to β-actin (expressed as arbitrary units). STAT activation was assessed with WB assays using phosphorylated (p)-STAT-specific antibodies. Alterations in STAT activation were calculated by normalizing p-STAT proteins to corresponding total STAT levels. Immunohistochemistry was performed on AAA and NA samples using the total and pSTAT antibodies.Systemic alterations in STAT activation were assessed by evaluating circulating leukocytes for the presence of p-STAT from patients with AAA (AAA, N=8), repaired aneurysm (RA, N=8), or age/gender matched controls with no AAA (CT, N=8). Flow cytometry was performed to assess for circulating levels of STAT1 (pY701), STAT3 (pY705), and STAT5a (pY694) in monocytes, granulocytes, and lymphocytes. Assessments were made at baseline and in response to in vitro stimulation with IFN-gamma (50 ng/mL) or IL-6 (100 ng/mL). Results were analyzed using Student’s T-test and are expressed as mean±SEM. In AAA tissue compared to NA, STAT-1 (1.08±0.09 v. 0.62±0.07), -2 (0.98±0.07 v. 0.55±0.08), and -4 (0.89±0.12 v. 0.35±0.11) mRNA levels were elevated (P<0.01, all). Corresponding increases in STAT protein were only observed for STAT1 (2.77±0.93 v. 0.93±0.08, P<0.05). Increases in activation were observed in AAA compared to NA in p-STAT2 (0.77±0.1 v. 0.1±0.02, P<0.01), p-STAT3 (1.6±0.3 v. 0.2±0.06, P<0.02) and p-STAT5 (0.57±0.03 v. 0.2±0.03, P<0.05) levels. Phosphorylated STAT1, 2, 3, and 5 were observed in inflammatory cells invading the AAA adventitia. In addition, STAT3 was observed in the media of AAA and NA, but pSTAT3 was only observed in the media of AAA. There were no differences in baseline levels of p-STAT-positive circulating leukocytes. IFN-gamma stimulation decreased STAT-5a (pY694)-positive CT lymphocytes to 40±13% of baseline, but had no effect on AAA or RA lymphocytes (116±35%, 102±19%, respectively; P=0.01). STAT-5a (pY694)-positive CT granulocytes also decreased to 62±18% of baseline compared to AAA or RA granulocytes (122±25%, 126±17%, respectively; P=0.01). Alterations in STAT1 (pY701) and STAT3 (pY705) were not observed in leukocytes following cytokine stimulation. STAT proteins are important regulators of transcriptional activity and have been linked to cardiovascular disease. The present data suggest that altered levels of phosphorylated STATs are associated with AAA. Understanding their role may provide further insight into the mechanisms of AAA formation and allow for the development of medical treatment options.
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影响因子：
32.4
作者：
Stritesky GL;Muthukrishnan R;Sehra S;Goswami R;Pham D;Travers J;Nguyen ET;Levy DE;Kaplan MH
通讯作者：
Kaplan MH
影响因子：
6
作者：
Lindeman, Jan H. N.;Abdul-Hussien, Hazem;Kleemann, Robert
通讯作者：
Kleemann, Robert
影响因子：
37.8
作者：
King, Victoria L.;Lin, Alexander Y.;Gerszten, Robert E.
通讯作者：
Gerszten, Robert E.
影响因子：
4.9
作者：
Carey, Rebecca;Jurickova, Ingrid;Denson, Lee A.
通讯作者：
Denson, Lee A.
DOI：
10.1158/1940-6207.capr-09-0105
发表时间：
2010-04
期刊：
Cancer prevention research (Philadelphia, Pa.)
影响因子：
--
作者：
Gamero AM;Young MR;Mentor-Marcel R;Bobe G;Scarzello AJ;Wise J;Colburn NH
通讯作者：
Colburn NH