The kallikrein-kinin system in humans

1. Kinin peptides are implicated in many physiological and pathological processes, including the regulation of blood pressure and sodium homeostasis, inflammation and the cardioprotective effects of preconditioning. In humans, the plasma and tissue kallikrein-kinin systems (KKS) generate bradykinin and kallidin peptides, respectively. 2. We established methodology for the measurement of bradykinin and kallidin peptides and their metabolites in order to study the function of the plasma and tissue KKS in humans. 3. Bradykinin peptides were more abundant than kallidin peptides in blood and cardiac atrial tissue, whereas kallidin peptides were predominant in urine. The levels of kinin peptides in tissue were higher than in blood, confirming the primary tissue localization of the KKS. 4. Angiotensin-converting enzyme inhibition increased blood levels of bradykinin and kallidin peptides. 5. Blood levels of kallidin peptides were suppressed in patients with severe cardiac failure, indicating that the activity of the tissue KKS is suppressed in this condition. 6. Bradykinin peptide levels were increased in the urine of patients with interstitial cystitis, suggesting a role for these peptides in the pathogenesis and/or symptomatology of this condition. 7. Cardiopulmonary bypass, a model of activation of the contact system, activated both the plasma and tissue KKS. 8. Measurement of individual bradykinin and kallidin peptides and their metabolites gives important information about the operation of the plasma and tissue KKS and their role in physiology and disease states.     

人血浆组织激肽释放酶结合蛋白Kallistatin与冠状动脉病变程度相关性研究

目的:分析组织激肽释放酶结合蛋白(kallistatin,KS)在冠心病患者血浆中水平的差异以及与Gensini评分的相关性,揭示血浆KS水平与冠状动脉病变严重程度的相关性?方法:选取151例行冠脉造影检查的患者作为研究对象,统计其临床数据及造影结果?①根据冠脉病变分支结果分成4组:正常对照组?单支病变组?双支病变组?三支病变组,应用ELISA法测定患者血浆中KS含量;②根据Gensini评分评估其冠脉病变程度,将患者分为4组:A组(0分)?B组(>0~≤20分)?C组(>20~≤40分)?D组(>40~?艽160分),两两比较各组间患者血中KS含量的差异,并探究KS水平与Gensini评分间的相关性?结果:①根据冠脉病变分支结果分的4组中,血浆KS量分别为:对照组(95.35 ± 32.00)μg/mL?单支病变组(78.19 ± 25.27)μg/mL?双支病变组(66.19 ± 25.93)μg/mL?三支病变组(51.84 ± 23.26)μg/mL,各组间均具有统计学差异(P < 0.01),KS水平与冠脉病变支数呈负相关;②根据Gensini评分分的A?B?C?D 4组中,KS含量分别为(101.66 ± 36.50)μg/mL?(77.73 ± 25.13)μg/mL?(61.58 ± 23.06)μg/mL?(53.80 ± 23.92)μg/mL,KS含量在各组间的差异有统计学意义(P <0.01);③KS含量与Gensini评分存在统计学上有显著的相关性(r=-0.371,P < 0.001)?结论:冠心病患者血浆KS含量明显低于正常人群,且KS含量与Gensini评分存在显著的相关性?提示KS可能作为一个潜在的血管保护因子参与动脉粥样硬化性心脏病的发生发展过程?     

多基因联合作用对kallistatin活性增强的研究

 目的 加强kallistatin(Kal)这一多功能内源性抗血管生成因子抗肿瘤活性。方法 将其他多个基因与Kal联合应用,利用脂质体将带有目的基因的质粒转染入内皮细胞和肿瘤细胞,分析多基因联合治疗对细胞特性的影响。结果 Kal对肺癌细胞A549、NCI-H446、SPC-A1都具抑制作用。Kal与Trail或vasostatin(Vas)联合,可加强对SPC-A1细胞的抑制作用,但三者联合该抑制作用并未进一步加强。Kal与angiostatin(Ang)、Vas 联合应用可显著增强对血管内皮细胞EVC304生长、迁移以及小管形成的抑制作用。结论 Kal基因可与多种类型的基因联合作用,增强对肿瘤细胞和血管内皮细胞的抑制作用,为肿瘤的多基因联合治疗提供了实验基础。     

Kallistatin抗炎作用的研究进展

Kallistatin(KS)是一种激肽释放酶结合蛋白(kallikrein-binding protein)，广泛分布于多种组织及体液中。KS不但具有抑制组织型激肽释放酶的作用，还具有抗炎、抗血管生成、抗肿瘤等多种生物学作用。KS生物学功能与其分子结构中不同的结构域有关。而KS抗炎作用被认为主要与肝素结合结构域有关。KS通过其肝素结合结构域，能竞争性抑制肿瘤坏死因子(tumor necrosis factor α，TNF-α)与它的受体的结合，从而起到影响TNF-α-NF-κB信号通路，发挥抗炎作用。KS也能与Kruppel样因子4(Kruppel-like Factor 4，KLF4)结合，激活内皮型一氧化氮合酶(endothelial nitric oxide synthase，eNOS)，抑制TNF-α活化核因子-κB(nuclear factor-κB，NF-κB)，发挥抗炎作用。但这与肝素结合结构域的机制不同。不同的机制可能为抗炎治疗提供新的思路，本文将就Kallistatin抗炎作用的相关研究进展进行阐述。     

Human kallistatin administration reduces organ injury and improves survival in a mouse model of polymicrobial sepsis

Kallistatin, a plasma protein, has been shown to exert multi‐factorial functions including inhibition of inflammation, oxidative stress and apoptosis in animal models and cultured cells. Kallistatin levels are reduced in patients with sepsis and in lipopolysaccharide (LPS)‐induced septic mice. Moreover, transgenic mice expressing kallistatin are more resistant to LPS‐induced mortality. Here, we investigated the effects of human kallistatin on organ injury and survival in a mouse model of polymicrobial sepsis. In this study, mice were injected intravenously with recombinant kallistatin (KS3, 3 mg/kg; or KS10, 10 mg/kg body weight) and then rendered septic by caecal ligation and puncture 30 min later. Kallistatin administration resulted in a > 10‐fold reduction of peritoneal bacterial counts, and significantly decreased serum tumour necrosis factor‐α, interleukin‐6 and high mobility group box‐1 (HMGB1) levels. Kallistatin also inhibited HMGB1 and toll‐like receptor‐4 gene expression in the lung and kidney. Administration of kallistatin attenuated renal damage and decreased blood urea nitrogen and serum creatinine levels, but increased endothelial nitric oxide synthase and nitric oxide levels in the kidney. In cultured endothelial cells, human kallistatin via its heparin‐binding site inhibited HMGB1‐induced nuclear factor‐κB activation and inflammatory gene expression. Moreover, kallistatin significantly reduced apoptosis and caspase‐3 activity in the spleen. Furthermore, kallistatin treatment markedly improved the survival of septic mice by 23% (KS3) and 41% (KS10). These results indicate that kallistatin is a unique protecting agent in sepsis‐induced organ damage and mortality by inhibiting inflammation and apoptosis, as well as enhancing bacterial clearance in a mouse model of polymicrobial sepsis.     

脓毒症休克患者血清Kal、VE-Cad的变化及与预后的关系

目的 探讨脓毒症休克患者血清人源性激肽释放酶结合蛋白（Kal）、血管内皮钙黏蛋白（VE-cad）的变化及与预后的关系。方法 前瞻性选取2016年3月—2018年3月杭州市丁桥医院重症医学科收治的脓毒症休克患者252例,根据纳入和排除标准排除74例,最终纳入研究的脓毒症休克患者为178例。根据患者28 d预后分为存活组120例和死亡组58例。比较两组患者的临床指标及入住重症监护病房（ICU）第1天、第3天、第5天血清Kal和VE-cad水平变化,记录患者急性生理学与慢性健康状况评分系统Ⅱ（APACHEⅡ）评分和序贯性器官功能衰竭评估（SOFA）评分。采用Pearson相关分析血清Kal、VE-cad与APACHEⅡ评分和SOFA评分的相关性。采用受试者工作特征（ROC）曲线评估血清Kal、VE-cad单独及联合检测对脓毒症休克患者预后的价值。采用二元Logistic回归分析脓毒症休克死亡的影响因素。结果 死亡组的氧合指数和血清白蛋白（ALB）水平低于存活组（P <0.05）,机械通气时间、ICU住院时间长于存活组（P <0.05）,血清C反应蛋白（CRP）、降钙素原（PCT）水平、血管外肺水指数（EVLWI）、APACHEⅡ评分和SOFA评分高于存活组（P <0.05）。两组入住ICU第1天、第3天、第5天血清Kal、VE-cad水平比较,不同时间点的血清Kal、VE-cad有差异（P <0.05）,两组血清Kal、VE-cad有差异（P <0.05）,两组血清Kal、VE-cad变化趋势有差异（P <0.05）。血清Kal水平与APACHEⅡ评分和SOFA评分呈负相关（r =-0.397和-0.466,P <0.05）;血清VE-cad与APACHEⅡ评分和SOFA评分呈正相关（r =0.173和0.192,P <0.05）。入院24 h内血清Kal、VE-cad及两者联合检测的曲线下面积（AUC）分别为0.687（95% CI：0.604,0.770）、0.859（95% CI：0.805,0.912）、0.890（95% CI：0.845,0.936）,血清Kal检测预测脓毒症休克患者死亡的敏感性为81.0%（95% CI：0.772,0.854）,特异性为50.0%（95% CI：0.474,0.531）;VE-cad检测预测脓毒症休克患者死亡的敏感性为69.0%（95% CI：0.649,0.726）,特异性为86.7%（95% CI：0.822,0.914）;血清Kal和VE-cad联合检测预测脓毒症休克患者死亡的敏感性为82.2%（95% CI：0.781,0.868）,特异性为89.6%（95% CI：0.852,0.954）。CRP［R=1.221（95% CI：1.022,1.459）］、PCT［R=1.195（95% CI：1.035,1.380）］、VE-cad［R=1.373（95% CI：1.055,1.787）］、EVLWI［R=1.846（95% CI：1.178,2.893）］是脓毒症休克患者死亡的危险因素（P <0.05）,Kal［R=0.428（95% CI：0.190,0.966）］是脓毒症休克患者死亡的保护因素（P <0.05）。结论 CRP、PCT、Kal、VE-cad、EVLWI是脓毒症休克患者死亡的影响因素,脓毒症休克患者血清VE-cad水平升高,Kal水平降低,两者的联合检测对评估脓毒症休克患者的预后有一定临床价值。