阿司匹林和氯吡格雷联合应用 对小鼠脑缺血恢复期RAGE及 sRAGE表达的影响

目的 探索阿司匹林和氯吡格雷联合应用对小鼠大脑中动脉远端缺血再灌注模型恢复期晚期糖基化终末产物受体（receptor for advanced glycation end products,RAGE）及可溶性晚期糖基化终末产物受体（soluble receptor for advanced glycation end products,sRAGE）表达的影响及其机制。 方法 60只C57BL/6J雄性小鼠随机分为假手术组、溶剂组、阿司匹林和氯吡格雷联合组（双抗组）,每组20只。通过压迫大脑中动脉远端制作脑缺血再灌注模型（缺血60?min再灌注）,再灌注即刻灌胃,溶剂组给予饮用水100?μL,双抗组给予阿司匹林（剂量12?mg/kg,每只实际用量0.3?mg）与氯吡格雷（剂量12?mg/kg,每只实际用量0.3?mg）混悬液100?μL,每日1次,连续21?d。缺血1、3、5、7、9、11、14、21?d对小鼠进行神经功能评价。缺血21?d取材,酶联免疫吸附法检测小鼠血清sRAGE表达水平;免疫印记检测脑组织RAGE表达水平;实时荧光定量PCR检测CD16、CD32、CD11b、诱导型一氧化氮合成酶（inducible nitric-oxide synthase,iNOS）、CD206、精氨酸酶1（arginase1,Arg1）、转化生长因子β（transforming growth factor β,TGF-β）、几丁质酶样蛋白（chitinase-like 3,Chil3/Ym1/2）等炎症因子及RAGE的mRNA表达情况;免疫荧光染色标记RAGE、神经元特异核蛋白（neuronal nuclei,NeuN）、胶质纤维酸性蛋白（glial fibrillary acidic protein,GFAP）,观察RAGE与神经元和星形胶质细胞共定位情况。 结果 双抗组小鼠3?d时胡须碰触评分和神经功能总分、9?d时神经功能总分优于溶剂组,差异有统计学意义（P=0.0067、0.0140、0.0406）。缺血再灌注21?d时,双抗组小鼠血清sRAGE表达水平高于溶剂组（1.099±0.541?ng/mL?vs.?0.319±0.341?ng/mL,P=0.0120）;脑组织iNOS（1.250±0.318 vs.?1.843±0.301,P=0.0164）及RAGE（2.105±0.300?vs.?2.732±0.249,P=0.0071）的mRNA相对表达较溶剂组下降。在小鼠脑梗死周边区RAGE与神经元具有共定位,与星形胶质细胞无共定位;双抗组RAGE+NeuN+细胞数量少于溶剂组（328.798±35.183个/平方毫米?vs.?814.437±165.758个/平方毫米,P=0.0012）。 结论 阿司匹林和氯吡格雷联合应用可能通过下调RAGE、上调sRAGE的表达水平,减轻炎症反应,从而发挥脑保护作用。     

sRAGE plays a role as a protective factor in the development of PCOS by inhibiting inflammation

Abstract

This study determined the effect of exogenous soluble receptor for advanced glycation end products (sRAGE) on the pro-inflammatory activities that occur during polycystic ovary syndrome (PCOS) in human follicular cells and explored a potential mechanism for preventing the development of inflammation. Follicular fluid was allocated into one of three treatment groups (0, 0.6, and 1.2?μg mL^?1 of sRAGE). Collectively, these results indicate that exogenous sRAGE supplementation alleviates inflammation in ovarian follicular granulosa cells by regulating p-ERK and AP-1 signaling.     

血浆S100A12与婴幼儿体外循环术后非感染性肺部并发症的相关性研究

目的 动态观察婴幼儿体外循环(CPB)围术期血浆中S100A12和可溶性晚期糖基化终末产物受体(sRAGE)水平变化,探讨其在婴幼儿CPB术后非感染性肺部并发症(NPC)早期预测中的作用.方法 采用病例对照研究.选取2011年6月1日至7月31日,在浙江大学附属儿童医院行CPB下心内直视矫治术、年龄＜3岁的先天性心脏病患儿.排除术前肺部有炎症性疾病和肝肾功能异常的患儿.根据术后是否出现胸腔积液、乳糜胸、部分肺不张、肺动脉高压危象、气道紊乱、气胸、纵膈积气、膈神经麻痹,20例被分为NPC组,40例被分为no-NPC组.于手术前,CPB前,CPB后,手术后1、12和24 h,采用ELISA法检测血浆中S100A12和sRAGE浓度.两组之间的浓度差异采用t检验;多因素Logistic回归分析检验S100A12和sRAGE在术后NPC早期预测中的作用,并以优势比(OR)及95％可信区间(95％ CI)表示相对危险度;以P＜0.05为差异具有统计学意义.结果 血浆中S100A12和sRAGE浓度在CPB后立刻明显升高(P＜0.01).术后24h后血浆中sRAGE水平明显低于术前水(P＜0.05),而S100A12仍明显高于术前水平(P＜0.01).CPB术后即刻,NPC组血浆中S100A12和sRAGE水平明显高于no-NPC组(P＜0.05).术后24 hNPC组血浆中S100A12水平仍明显高于no-NPC组(P＜0.05),而两组患儿血浆中sRAGE水平无明显差异(P＞0.05).逐步法Logistic回归分析,CPB后即刻血浆中S100A12水平与术后NPC的发生明显相关(OR=1.042,95％ CI:1.010～ 1.076,P=0.011).CPB后即刻血浆中S100A12与术后机械通气时间(r=0.47,P＜0.01)、住ICU时间(r=0.363,P=0.008)和住院时间(r =0.402,P=0.002)明显相关.结论 婴幼儿CPB术后血浆中S100A12和sRAGE水平立刻明显升高.血浆中S100A12是婴幼儿CPB术后NPC发生和预后的早期预测指标.     

Circulating levels of advanced glycation end products (AGE) and interleukin-6 (IL-6) are independent determinants of serum asymmetric dimethylarginine (ADMA) levels in patients with septic shock

There is a growing body of evidence that nitric oxide (NO) excess plays a central role in the pathogenesis of hypotension and organ failure in patients with septic shock. In addition, recently, asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NO synthase, has been shown to contribute to the regulation of vascular tone via modulation of NO generation in vivo. However, the kinetics and regulation of serum levels of ADMA in patients with septic shock are largely unknown. Since high mobility group box 1 (HMGB1)-receptor for advanced end products (RAGE) axis is supposed to be involved in the lethality in septic shock, we examined the correlations among serum levels of ADMA, endotoxin, interleukin-6 (IL-6), soluble form of RAGE (sRAGE) and RAGE ligands such as HMGB1 and advanced glycation end products (AGE) in septic shock patients. Fifteen septic shock patients (10 males and 15 females, mean age: 70.1 ± 8.5 years) and fifteen age- and sex-matched healthy volunteers were included in this study. The criteria of the American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference were used for diagnosis of septic shock. All the subjects underwent a complete history and physical examination, determination of blood chemistries, including serum levels of ADMA, endotoxin, IL-6, HMGB1, AGE and sRAGE. Linear and multiple stepwise regression analysis were performed for the determinants of serum levels of ADMA. Serum levels of ADMA were significantly higher than those in healthy volunteers (0.98 ± 0.21 nmol/mL vs. 0.30 ± 0.05 nmol/mL, p < 0.0001). In univariate analysis, creatinine (p < 0.005), endotoxin (p < 0.001), IL-6 (p < 0.001), HMGB1 (p < 0.001), AGE (p < 0.001) and sRAGE (p < 0.001) were significantly associated with serum ADMA levels. After performing multivariate stepwise regression analyses, IL-6 (p = 0.001), AGE (p = 0.002) and creatinine (p = 0.013) still remained significant independently. The present study is the first demonstration that ADMA levels were significantly elevated in patients with septic shock and that serum IL-6, AGE and creatinine levels were independent determinants of ADMA in these patients. Given the harmful effects of NO excess in septic shock, ADMA levels may be increased as a counter-system against inflammation and oxidative stress in this life-threatening disorder.     

慢性阻塞性肺疾病患者sRAGE与AGEs水平变化及意义

目的探讨慢性阻塞性肺疾病患者血浆sRAGE与AGEs水平变化及意义。方法选取我院80例慢阻肺急性加重期患者与80例慢阻肺稳定期患者,分别测量其血浆sRAGE与AGEs浓度。结果慢阻肺急性加重期患者的血浆AGEs浓度明显高于慢阻肺稳定期患者(P0.01);而血浆sRAGE浓度明显低于稳定期患者(P0.01);Pearson相关分析结果显示,血浆AGEs浓度与FEV1%呈负相关,血浆sRAGE浓度与FEV1%呈正相关。结论血浆AGEs可能是慢阻肺的危险因素,血浆sRAGE则可能是慢阻肺的保护因素,两者对判断慢阻肺病情严重程度及预后具有一定的应用价值。