内皮素-1对大鼠血管平滑肌细胞一氧化氮及硫化氢体系的影响

目的探讨不同浓度内皮素-1（ET-1）对大鼠血管平滑肌细胞（A7r5细胞株）内源性一氧化氮（NO）及硫化氢（H2S）体系的影响。方法将A7r5细胞分为对照组和实验组，在实验组中分别加入浓度为10－8～10－6 mol/L的ET-1，对照组加入等体积的无菌磷酸盐缓冲液（PBS），48 h后分别应用NO和H2S探针原位检测A7r5细胞中NO及H2S水平，分别采用NO测定试剂盒和敏感硫电极法检测细胞上清中NO及H2S水平，应用Western blot法检测A7r5细胞中诱导型一氧化氮合酶（NOS2）、内皮型一氧化氮合酶（NOS3）、胱硫醚-γ-裂解酶（CSE）、胱硫醚-β-合成酶（CBS）及细胞增殖核抗原（PCNA）的表达。结果10－8、10－7及10－6 mol/L ET-1处理的A7r5细胞内NO水平的相对荧光强度（0.078±0.080、0.075±0.002、0.056±0.009）均低于对照组（0.094±0.061），差异有统计学意义（F＝15.248，P〈0.05）；细胞上清中NO水平[（1.391±0.134） μmol/L、（1.219±0.280） μmol/L、（1.116±0.181） μmol/L]均明显低于对照组[（2.131±0.484） μmol/L]，差异有统计学意义（F＝20.833，P〈0.01）；NOS2 （0.457±0.097、0.462±0.116、0.438±0.180）明显低于对照组（0.721±0.222），差异有统计学意义（F＝6.196，P〈0.05）；NOS3蛋白表达差异无统计学意义（F＝2.669，P〉0.05）。10－8、10－7、10－6 mol/L ET-1组A7r5细胞内H2S水平的相对荧光强度（0.063±0.002、0.056±0.008、0.042±0.009）低于对照组（0.082±0.006），差异有统计学意义（F＝16.297，P〈0.01）；细胞上清中H2S水平[（19.961±3.428） μmol/L、（17.516±5.144） μmol/L、（14.481±4.885） μmol/L]均明显低于对照组[（29.439±4.236） μmol/L]，差异有统计学意义（F＝12.518，P〈0.01）；CBS蛋白表达（0.359±0.096、0.270±0.038、0.174±0.051）均低于对照组（0.707±0.107），差异有统计学意义（F＝20.833，P〈0.01），CSE蛋白表达差异无统计学意义（F＝0.708，P〉0.05）。10－7 mol/L ET-1组细胞PCNA表达高于对照组（0.686±0.180比0.437±0.191），差异有统计学意义（t＝－2.840，P〈0.01）。结论ET-1刺激可导致血管平滑肌细胞增殖，并使其内源性NO和H2S体系下调。

内源性二氧化硫对氯化钴诱导人肺动脉内皮细胞凋亡的影响

目的研究内源性二氧化硫（SO2）对氯化钴（CoCl2）诱导人肺动脉内皮细胞（HPAECs）凋亡的调节作用。方法采用CoCl2诱导原代HPAECs制备低氧刺激细胞凋亡模型，采用慢病毒转染过表达内源性SO2生成关键酶天冬氨酸氨基转移酶1（AAT1），HPAECs分为4组：空载体（vehicle）组、vehicle＋CoCl2组、AAT1组及AAT1＋CoCl2组。采用Western blot法检测AAT1、B细胞淋巴瘤/白血病-2（bcl-2）、bcl-2相关X蛋白（bax）、Caspase-3蛋白和活化的Caspase-3（cleaved Caspase-3）的表达，采用比色法检测AAT活性，采用荧光探针原位检测HPAECs中SO2水平，采用依赖于末端脱氧核苷酸转移酶的dUTP末端标记法（TUNEL）测定细胞凋亡。结果4组HPAECs中SO2含量、AAT1和bcl-2蛋白表达及cleaved Caspase-3/Caspase-3比值差异均有统计学意义（F＝147.364、23.738、6.521、64.884，均P〈0.05），但bax蛋白表达差异无统计学意义（F＝1.620，P〉0.05）。与vehicle组相比，vehicle＋CoCl2组HPAECs中AAT活性[（0.96±0.24） Carmen′s unit/μg比（2.21±0.60） Carmen′s unit/μg]、SO2水平（40.71±7.72比105.60±16.20）和bcl-2蛋白表达（0.59±0.19比1.02±0.20）均显著下降，细胞凋亡和cleaved Caspase-3/Caspase-3比值（1.56±0.25比0.95±0.13）明显增加，差异均有统计学意义（均P〈0.05）；但AAT1蛋白表达差异无统计学意义（0.50±0.12比0.53±0.11，P〉0.05）。与vehicle组相比，AAT1组HPAECs中SO2水平（351.50±42.43比105.60±16.20）和AAT1蛋白表达（1.22±0.33比0.53±0.11）均显著升高，差异均有统计学意义（均P〈0.05）。与AAT1组相比，AAT1＋CoCl2组HPAECs中SO2水平（333.50±46.22比351.50±42.43）、AAT1蛋白表达（1.26±0.36比1.22±0.33）和bcl-2蛋白表达（1.14±0.38比1.03±0.27）差异无统计学意义，细胞凋亡和cleaved Caspase-3/Caspase-3比值（0.51±0.17比0.50±0.11）差异无统计学意义（均P〉0.05）。结论内源性SO2可抑制CoCl2诱导HPAECs凋亡。

Intermedin/adrenomedullin2: an autocrine/paracrine factor in vascular homeostasis and disease

Intermedin(IMD)or adrenomedullin 2 is a novel peptide related to the calcitonin gene-related peptide(CGRP)family.Via calcitonin receptor-like receptor/receptor activity modifying proteins,the common receptor complexes of CGRP,IMD exerts a wide range of biological effects,especially regulation of cardiovascular homeostasis.Proteolytic processing of a larger IMD precursor yields a series of biologically active C-terminal fragments,IMD1–53,IMD1–47 and IMD8–47.IMD and its receptors are present in the cardiovascular system,and IMD is present at low levels in plasma.In the cardiovascular system,IMD has multiple functions such as regulation of blood pressure and cardiac function,pro-angiogenesis,endothelial barrier function protection,anti-oxidative stress,and anti-endoplasmic reticulum stress.IMD participates widely in the pathogenesis of atherosclerosis,hypertension,pulmonary arterial hypertension and vascular calcification.It is a vascular regulatory factor of homeostasis and a vital endogenous protective factor against vascular diseases.

Immune regulation of hydrogen sulfide in children with acute lymphoblastic leukemia

Background Acute lymphoblastic leukemia （ALL） and chemotherapy can cause immune imbalance,and gaseous molecule hydrogen sulfide （H2S） can participate in the process of immune response.This study aimed to investigate the immune regulation of H2S in pediatric ALL.Methods Children （n=78） with ALL admitted during 2010-2013 were included in this study.Two blood samples were collected in period of before chemotherapy,bone marrow remission and two days after chemotherapy,respectively.Serum contents of H2S and cytokines,including interleukin-1β （IL-13）,interleukin-2 （IL-2）,interferon-γ （IFN-γ）,tumor necrosis factor （TNF-α）,interleukin-4 （IL-4）,interleukin-6 （IL-6）,interleukin-10 （IL-10） and macrophage inflammatory protein-1α （MIP-1a）,were detected using ELISA method.Stepwise regression was used to analyze the correlation between H2S and cytokines.Furthermore,human Jurkat cells were cultured in vitro,and nucleoprotein of Jurkat cells and peripheral blood mononuclear cells （PBMCs） were collected,contents of cystathionine γ-lyase （CSE） and certain cytokines were measured by Western blotting.Results Serum concentrations of H2S,IL-13,IL-6,IL-10 and MIP-1α in children with ALL were increased significantly （P ＜0.01）,while concentrations of IL-2,TNF-α,IFN-γ and IL-4 decreased obviously （P ＜0.01）.In patients after chemotherapy,concentrations of H2S and IL-10 were decreased significantly （P ＜0.05）,but IL-4 and IFN-γ concentrations increased markedly （P ＜0.05）.At remission stage,H2S,IL-13,IL-4,IL-6,IL-10 and MIP-1α concentrations were further decreased markedly （P ＜0.05）,but concentrations of IL-2,TNF-α and IFN-γ increased again （P ＜0.05）.Protein contents of CSE,IL-10,IL-4 and IL-2 of PBMCs also increased markedly in children with ALL.Moreover,changes of CSE protein contents of PBMCs were consistent with serum H2S contents,and there were significant correlation between H2S and certain cytokines based on stepwise regression analysis.Furthermore,compared with those of PBMCs group,in vitro study indicated that Jurkat cells of H2S group expressed IFN-γ,IL-10,IL-4 and IL-2 protein increased obviously （P ＜0.05）,while IL-4,IL-2 and CSE expression of PPG group decreased markedly （P ＜0.05）.Conclusion Gaseous molecule H2S might participate in the process of immune regulation in pediatric ALL through modulating transcription and expression of cytokines.

Changes of atrial natriuretic peptide and antidiuretic hormone in children with postural tachycardia syndrome and orthostatic hypertension： a case control study

Background The abnormal blood volume regulation is one of the most important pathogenesis in postural tachycardia syndrome in children.This study was designed to investigate the plasma atrial natriuretic peptide and antidiuretic hormone levels in postural tachycardia syndrome children,and their associations with the changes in heart rate and blood pressure in head-up test.Methods Twenty-one postural tachycardia syndrome patients （（12±2） years） and 26 healthy children （（12±1） years） were included.According to blood pressure changes in head-up test,the postural tachycardia syndrome patients were divided into two subgroups：postural tachycardia syndrome with orthostatic hypertension and postural tachycardia syndrome without orthostatic hypertension.The plasma atrial natriuretic peptide and antidiuretic hormone levels were measured using enzyme-linked immunosorbent assay.Results The plasma atrial natriuretic peptide level in postural tachycardia syndrome patients was higher than the control （P=0.004）,whereas the difference in plasma antidiuretic hormone level between postural tachycardia syndrome and controls was not significant （P=0.222）.The plasma antidiuretic hormone level of patients suffering from postural tachycardia syndrome with orthostatic hypertension was much higher than that of children having postural tachycardia syndrome without orthostatic hypertension （P ＜0.05）.In postural tachycardia syndrome patients,the updght max heart rate was positively correlated with the plasma atrial natriuretic peptide level （r=0.490,P＜0.05） and the upright systolic blood pressure was positively correlated with the plasma antidiuretic hormone levels （r=0.472,P ＜0.05）.Conclusions There was a disturbance of plasma atrial natriuretic peptide and antidiuretic hormone in postural tachycardia syndrome children.

Adrenomedullin inhibits the endothelin production induced by urotensin Ⅱ in rat vascular smooth muscle cells

The aim of this study was to observe the effects of adrenomedullin (ADM) on endothelin (ET) production induced by urotensin Ⅱ (UⅡ) in rat vascular smooth muscle cells (VSMCs). Cultured VSMCs which were incubated with UⅡ (10-8 mol/L) and with various concentrations of ADM were used to measure the VSMCs 3H-TdR incorporation, the activity of extracellular signal-regulated kinase (ERK), the amount of ET mRNA and ET production in VSMCs. In this work we found that incubation with UⅡ (10-8 mol/L) increased obviously the amount of ET mRNA in VSMCs and ET production in medium, however, co-incubation with ADM (10-10-10-8 mol/L) and UⅡ(10-8 mol/L) reduced the amount of ET mRNA by 15%, 24% and 45% (P【 0.01) respectively, compared with UⅡ alone. The content of ET in medium was 14.13, 11.38 and 11.00 pg/mL. ADM alone (10-8 mol/L) had no effect on ET production in VSMCs. UⅡ (10-8 mol/L) promoted the 3H-TdR incorporation and activity of ERK in VSMCs. ADM inhibited VSMCs 3H-TdR incorporation and activation of