The role of calcineurin in the lung fibroblasts proliferation and collagen synthesis induced by basic fibroblast growth factor

Objective To investigate the role of calcineurin (CaN) in the lung fibroblast proliferation and collagen synthesis induced by basic fibroblast growth factor (bFGF).Methods We used Western blot and immunohistochemical methods for investigating the content and distribution of calcineurin in the lung tissue. Calcineurin activity in different tissues was measured using 32P-labelled substrate. In the primary culture of lung fibroblasts, 3H-thymidine (3H-TdR) and 3H-proline incorporation methods were used to study the effect of cyclosporin A (CsA), an inhibitor of calcineurin, on the lung fibroblast DMA and collagen synthesis stimulated by bFGF.Results We found that calcineurin was expressed in lung tissue and has phosphatase activity (7.1±2.0 pmol Pi/mg pr/min). CsA (10~(-8)-10~(-6) mol/L) inhibited lung fibroblast 3H-TdR incorporation induced by bFGF in a dose-dependent manner, with the inhibitory rates by 20% , 46% and 66% (P< 0.01). CsA (10~(-7) -10~(-6) mol/L) inhibited 3H-proline incorporation in lung     

全反式维甲酸对培养大鼠主动脉血管平滑肌细胞增生及P21表达的影响

目的　探讨全反式维甲酸 (ATRA)对培养大鼠主动脉血管平滑肌细胞增生及P2 1蛋白表达水平的影响。方法　血管平滑肌细胞 (VSMC)采用组织贴块法培养。ATRA(2 .5×10 -6mol·L-1)作用于经 2 0 %胎牛血清刺激后的血管平滑肌细胞 ,2 4h后分别行 3H TdR掺入实验测定和P2 1蛋白印迹检测P2 1蛋白表达。结果　ATRA明显抑制血管平滑肌细胞DNA合成和促进P2 1蛋白表达。结论　ATRA促进P2 1蛋白表达是抑制VSMC增生的原因之一     

降钙素基因相关肽对尾加压素Ⅱ诱导的血管平滑肌增殖的影响

目的 :观察降钙素基因相关肽 (CGRP)对尾加压素Ⅱ (UrotensionⅡ ,UⅡ )刺激的血管平滑肌细胞 (VSMC)增殖的影响及机制。方法 :贴块法培养大鼠胸主动脉VSMC ;3 H -胸腺嘧啶 (3 H -TdR)掺入测定VSMCDNA合成 ;γ- 3 2 P -ATP标记的同位素法测定丝裂素活化蛋白激酶 (MAPK)活性。结果 :UⅡ (10 -8mol/L)显著促进VSMC3 H -TdR掺入和激活MAPK。与对照组比较 ,分别增加 4 8%和 2 2 6 % (P <0 .0 1)。CGRP有效抑制UⅡ诱导的VSMC3 H -TdR掺入和MAPK激活。与UⅡ组比较 10 -9、10 -8、10 -7mol/LCGRP分别使VSMC3 H -TdR掺入降低 18%、2 5 %和31% (P <0 .0 1) ,使MAPK活性分别降低 2 6 %、5 0 %和 6 4 % (P <0 .0 1)。结论 :CGRP抑制UⅡ诱导的VSMC增殖 ,其机制可能与CGRP拮抗UⅡ刺激的MAPK活性有关     

Effect of arsenic trioxide on inhibition of restenosis after rabbit vascular injury and its mechanism

目的　探讨应用三氧化二砷 (As2 O3)预防血管损伤后再狭窄的效果及其作用机理。方法　观察As2 O3对培养兔血管平滑肌细胞 (VSMCs)凋亡的影响。 32只新西兰白兔随机分为 2、4周实验组和对照组 ,分别 10 %As2 O32 5mg·Kg 1·d 1或等量生理盐水腹腔注射 3天 ,应用球囊损伤左颈总动脉。处死动物取血管作形态学和免疫组化检测 ,并检查肝脏、肾脏组织学变化。结果　细胞形态学和DNA电泳梯形带证实 ,As2 O3诱导培养VSMCs凋亡 ,与药物浓度和作用时间呈依赖性。与对照组相比 ,2 wk实验组血管内膜增殖面积显著减少 (P <0 0 5 ) ,4 wk组内膜面积无明显差异 ;但 2 ,4 wk组的管腔面积均有明显增大 (P均 <0 0 5 )。与对照组相比 ,实验组 2 ,4 wk免疫组化显示bcl 2表达下调 (P均 <0 0 5 ) ,Bax表达上调 (P分别 <0 0 1,0 0 5 ) ,均与相对应的血管内膜增生受抑制 ,血管腔面积扩大相吻合结论　As2 O3诱导VSMCs凋亡和有效预防实验性血管损伤后再狭窄 ,均与下调bcl 2和上调bax表达有关     

舒芬太尼对大鼠腹主动脉平滑肌细胞钙激活钾电流的影响

目的:观察舒芬太尼对大鼠腹主动脉平滑肌细胞(AASMCs)钙激活钾电流(IKCa)的影响,探讨其扩张血管的作用机制。方法:用酶消化法急性分离AASMCs,采用全细胞膜片钳技术记录IKCa,观察不同浓度(1×10-8、3×10-8、1×10-7mol/L)舒芬太尼对该电流的影响。结果:与不加药对照组相比舒芬太尼能显著增大IKCa幅度(P<0.05或P<0.01),这一作用具有可逆性,并具有浓度依赖性。结论:舒芬太尼能增强AASMCs钙激活钾通道的活动,这一作用可能与临床上观察到的舒芬太尼舒血管效应有关。     

血管活性肽对同型半胱氨酸刺激平滑肌细胞增殖的实验研究

目的 观察对兔主动脉平滑肌细胞（ＶＳＭＣ）增殖起负调节作用的Ｃ－型利钠利尿肽（ＣＮＰ）、肾上腺髓质素（Ａｄｍ）、降钙素基因相关肽（ＣＧＲＰ）、生长抑素（ＳＳＴ）和甲状旁腺素相关蛋白（ＰＴＨｒＰ）等活性肽对同型半胱氨酸（Ｈｃｙ）刺激的ＶＳＭＣ增殖的影响。方法 ６只大耳白兔胸主动脉贴块法离体培养平滑肌细胞;分对照及Ｈｃｙ加不同处理因素组;＾３Ｈ－ＴｄＲ掺入测定ＶＳＭＣ增殖;差速离心分离细胞膜、胞浆及胞     

L-精氨酸及牛磺酸对溶血性磷脂酸诱导的血管平滑肌增殖的作用

目的 :观察牛黄酸 (TAU)及L -精氨酸 (L -Arg)对溶血性磷脂酸 (LPA)诱导的血管平滑肌 (VSMC)增殖的影响并探讨其可能的机制。方法 :贴块法培养大鼠胸主动脉VSMC ;3H -胸腺嘧啶 (3H -TdR)掺入测定VSMCDNA合成 ;γ - 32 P -ATP标记的同位素法测定丝裂素活化蛋白激酶 (MAPK)活性。结果 :与对照组相比 ,LPA(10 - 8mol L)可分别使VSMC3H -TdR掺入和MAPK活性增高 5 8%和 16 4 % (P <0 .0 1)。L -Arg及TAU均能浓度依赖性地抑制LPA诱导的VSMC3H -TdR掺入和MAPK激活。混合应用牛磺酸和L -精氨酸对LPA诱导的VSMC增殖及MAPK活性的抑制作用显著大于单用L -Arg和TAU(P <0 .0 1)。结论 :L -Arg及TAU均能抑制LPA诱导的VSMC增殖和MAPK激活 ,而合用L -Arg及TAU对抑制LPA诱导的VSMC增殖及MAPK活性具有协同和叠加效应     

全反式维甲酸对培养大鼠主动脉血管平滑肌细胞增殖及细胞周期素E表达的影响

目的　研究全反式维甲酸 (ATRA)对培养大鼠主动脉血管平滑肌细胞 (VSMC)增殖及细胞周期素E(Cy clinE)蛋白表达水平的影响。方法　血管平滑肌细胞采用组织贴块法培养。ATRA(2 .5× 10 - 6 mol·L- 1 )作用于经 2 0 %胎牛血清刺激后的VSMC ,2 4h后分别行3H TdR掺入实验测定和蛋白印迹检测CyclinE蛋白表达。 结果　ATRA明显抑制ATRA的DNA合成 (P <0 .0 1)和CyclinE蛋白的表达 (P <0 .0 5 )。结论　ATRA抑制CyclinE蛋白的表达是抑制VSMC增殖的原因之一。     

辛伐他汀对鼠心脏成纤维细胞DNA合成的影响

目的探讨辛伐他汀(Sim)对新生SD大鼠心脏成纤维细胞(CFs)DNA合成的影响及甲羟戊酸(MVA)的干预效应。方法采用胰酶消化、差速贴壁法培养新生SD大鼠CFs,以3H-胸腺嘧啶核苷(3H-TdR)掺入法测定DNA合成,观察不同浓度Sim及MVA分别作用不同时间对CFsDNA合成功能的影响。结果 (1)CFs的3H-TdR掺入率随着Sim干预浓度的增加而降低,其中10-6和10-5mol／L Sim组的3H-TdR掺入率分别为(1175±202．66)、(771±164．86)cpm／2000细胞, 显著低于对照组的(1608±204．32)cpm／2000细胞(均P<0．01);(2)10-5mol／L Sim作用CFs 6、12、18、24、30、36、42、48 h, 随着Sim作用时间的延长,CFs的3H-TdR掺入率呈递减趋势,与时间呈明显的负相关(r=-919,P<0．01);(3)CFs的3H-TdR 掺入率随着MVA干预浓度的增加呈进行性上升,其中10-4、10-3 mol／L MVA 10-5 mol／LSim组3H-TdR掺入率分别为 (1612±308．57)、(1995±353．83)cpm／2000细胞,显著高于对照10-5mol/L Sim组的(771±164．86)cpm／2000细胞(P<0．01); (4)10-3 mol／L MVA分别作用CFs 6-48 h,CFs的3H-TdR掺入率随着MVA作用时间的延长而逐渐增加,呈明显的正相关(r=0．968,P<0．01)。结论 Sim呈浓度-时间依赖方式抑制CFs DNA的合成且可被MVA拮抗,提示Sim可抑制CFs 增殖,延缓心肌纤维化,其作用可能通过MVA途径实现。     

Interaction of hypertension and diabetes on impairment of endothelial function

目的　评价血管平滑肌功能 (硝酸甘油引起的血管舒张 )不全与内皮功能不全的联系并探讨糖尿病与高血压对内皮功能的损害是否具有协同作用。方法　用高分辨率超声波仪测定肱动脉对硝酸甘油 (NTG)的扩张反应及血流介导的内皮依赖性血管舒张(EDD)。结果　平滑肌功能受损与受损的EDD呈正相关 (r =0 54,P <0 0 0 1)。单纯糖尿病或高血压仅EDD较对照组降低 ( 5 74 %± 3 32 % ;4 14%± 2 93%对 9 4 5%± 3 88% ,P分别 <0 0 5及 0 0 1) ,糖尿病合并高血压组不仅EDD受损 ( 2 78%± 2 0 8% ,P <0 0 0 1) ,平滑肌功能也受损 ( 14 11%± 4 63对 2 3 53%± 6 77% ,P <0 0 0 1)。结论　导致血管内皮功能不全的病理过程可能也同样损害血管平滑肌功能。 2型糖尿病与高血压相互作用加重血管舒张功能不全。     

Decreased small arterial compliance with increased serum vascular endothelial growth factor-A and circulating endothelial progenitor cell in dilated cardiomyopathy

Background Evidence showed that both myocardium and blood vessels were damaged in dilated cardiomyopathy （DCM）. However, the changes in arterial compliance, serum cytokines and circulating endothelial progenitor cells （EPC）, and their correlations remain unknown.
Methods Sixty-five DCM patients and 49 healthy volunteers were studied. Both large artery compliance （C1） and small artery compliance （C2） were measured with the CVProfUor DO-2020. Quantitative enzyme-linked immunosorbent assays （ELISAs） were used to measure the levels of vascular endothelial growth factor-A （VEGF-A） and VEGF receptor 2 （VEGF-R2）. Circulating EPC was assessed by EPC colony-forming assays and flow cytometry （CD133^＋/CD34^＋cells）. Phagocytized Dil-acLDL and binded FITC-UEA-I were used to analyze endothelial lineage marker expression by immunofluorescence.
Results Although C2 was markedly lower in DCM patients than in control group （（3.8±1.8） ml/mmHg × 100 vs （5.0±2.2） ml/mmHg × 100, P〈0.0001）, there was no statistically significant difference in C1 between the two groups （P〉0.05）. Levels of VEGF-A, the numbers of colony-forming units （CFU） and the fractions of EPC were obviously higher in DCM patients than in control group （（127.6±139.5） pg/ml vs （58.8±42.9） pg/ml, P〈0.0001; （2.5±1.5）% vs （0.5±0.3）%, P〈0.05; 23.5±12.8 vs 10.8±7.4, P〈0.01, respectively） and however, there was no significant difference in VEGF-R2 between two groups （P〉0.05）. LgVEGF-A was positively correlated with the number of EPC-CFU （r=-0.435; P〈0.05） and inversely correlated with C2 （r=-0.543; P〈0.001） in DCM patients. Conclusions The reduction of C2, a sensitive marker reflecting endothelial dysfunction, was observed in DCM patients and closely related to the increase in serum VEGF-A.     

Imbalance of endogenous homocysteine and hydrogen sulfide metabolic pathway in essential hypertensive children

Background  Hypertension is a common disease of the cardiovascular system. So far, the pathogenesis of primary hypertension remains unclear. The elaboration of its pathogenesis is an important topic in the field which calls for urgent resolution. The aim of this study was to probe into the metabolic imbalance of homocysteine (Hcy) and hydrogen sulfide (H₂S) in children with essential hypertension, and its significance in the pathogenesis of essential hypertension.
Methods  Twenty-five children with essential hypertension and 30 healthy children with normal blood pressure were enrolled in the study. The medical history was investigated and a physical examination was conducted on the subjects. Plasma Hcy content was examined by fluorescence polarization immunoassay (FPIA). The plasma H₂S level was detected by a modified method with a sulfide electrode. Data were presented as mean±standard deviation. The t test was applied to the mean values of both groups. Pearson linear correlation analysis was applied to the plasma Hcy and H₂S as well as to the systolic pressure against the plasma H₂S/Hcy ratio.
Results  Plasma Hcy, an intermittent metabolite of the endogenous methionine pathway, was markedly increased but plasma H₂S, a final product of this pathway was significantly decreased in hypertensive cases when compared with normal subjects ((Hcy: (12.68±9.69) µmol/L vs (6.62±4.79) µmol/L (t=2.996, P<0.01); H₂S: (51.93±6.01) µmol/L vs (65.70±5.50) µmol/L) (t=-8.670, P<0.01)). The ratio of plasma H₂S/Hcy in children with hypertension was 5.83±2.91, while that of the control group was 11.60±3.30, and the difference is significant with a t=-6.610 and P<0.01. A negative correlation existed between plasma Hcy and H₂S concentrations, r=-0.379, P<0.05. And a negative correlation was found between systolic blood pressure and the plasma H₂S/Hcy ratio, r=-0.687, P<0.05.
Conclusion  There was a metabolic imbalance of homocysteine and hydrogen sulfide in essential hypertensive children.

     

ATP敏感性钾通道开放对内皮素-1诱导平滑肌细胞增殖和凋亡的作用

目的:探讨ATP敏感性钾通道开放剂拉马克啉(Levcromakalim,Lev)对内皮素-1(ET-1)诱导的血管平滑肌细胞(VSMCs)增殖及凋亡的影响。方法:体外培养Wistar大鼠主动脉VSMCs,用ET-1诱导其增殖,用不同浓度拉马克啉共培养,MTT法评价VSMCs增殖情况,3H-TdR检测DNA合成;流式细胞术检测VSMCs凋亡。免疫细胞化学检测Bcl-2/Bax表达。结果:①Lev显著抑制对ET-1诱导的VSMCs增殖,呈浓度依赖效应,Lev组MTT细胞活性含量和3H-TdR掺入量都明显低于ET-1组(P<0.05)。②Lev组VSMCs凋亡较ET-1组明显增多(P<0.05);与对照组比较,Lev组细胞凋亡率也显著增加(P<0.01)。③Lev使VSMCs Bcl-2表达下调,Bcl-2/Bax比率下降。结论:ATP敏感性钾通道开放可抑制ET-1诱导的VSMCs增殖作用,通过调节Bcl-2/Bax表达增加VSMCs凋亡。     

The relationship between serum interleukins and T-lymphocyte subsets in patients with severe acute respiratory syndrome

Severeacuterespiratorysyndrome (SARS) ,alsoknownasinfectiousatypicalpneumonia (IAP)inChina ,iscausedbyacoronavirusmutant Thisrespiratorycontagiousdiseaseistransmittedmainlybyrespiratorydropletswithinaneardistanceandclosecontactwiththepatients Sofar ,thediseasehasbeenreportedinover 30countriesworldwide SARSpatientshavebeenadmittedintoourhospitalsinceMarch 11,2 0 0 3 Inthispaper ,wepresentthelaboratoryresultsofseruminterleukins (IL) ,T lymphocytesubsets ,whitebloodcell (WBC)countsof 35pa…     

Harm Reduction - Good Practice in Asia. The Integration of Harm Reduction into Abstinence-based Therapeutic Communities

Muscle atrophy may occur in aging skeletal muscles, neurodegenerative syndromes,disease related cachexia and spaceflight.1,2 It is characterized by a loss of body mass, mainly skeletal muscles. Metabolic abnormalities develop in various chronic diseases and lead to progressive catabolism with decrements in the skeletal musculature that result in muscle atrophy.2prevent cell death and restore muscle function.4 Circulating and muscular IGF-1 levels are frequently reduced in catabolic states.5 …     

宽叶缬草对动脉平滑肌细胞收缩及生长的影响

目的 :观察宽叶缬草 (VOL)对培养的血管平滑肌细胞 (VSMC)收缩及生长的影响。方法 :取 4～ 6周wistar大鼠胸主动脉中层平滑肌细胞进行原代及传代培养。观察VOL和L -NAME对VSMC收缩的影响 ,以及血管紧张素Ⅱ (AngⅡ )和不同浓度VOL时VSMC的3H -TdR及3H -Leucine参入变化。结果 :VOL可显著抑制AngⅡ引起的VSMC收缩 ,且此作用不受L -NAME影响。VOL呈剂量依赖性抑制VSMC的3H -TdR和3H -Leucine的参入。结论 :VOL有显著抑制AngⅡ引起的VSMC收缩和生长的作用     

胃促生长素对人主动脉平滑肌细胞增殖及线粒体融合蛋白2表达的影响

目的 通过体外培养人主动脉平滑肌细胞(HASMCs),研究胃促生长素(ghrelin)对血管平滑肌细胞(VSMC)增殖及线粒体融合蛋白2(Mfn-2)表达的影响.方法 体外培养HASMCs,第4～6代细胞用于试验.给予不同浓度(10-9、10-8、10-7、10-6、10-5 mol/L)的ghrelin 或10-6 mol/L ghrelin不同时间(0、6、12、18、24 h)处理,用四甲基偶氮唑蓝比色(MMT)法观察其对HASMC增殖的影响.RT-PCR,Western blot方法检测不同处理方法对Mfn-2表达的影响.结果 10-7～10-5 mol/L的ghrelin可明显抑制HASMC增殖,浓度为10-6 mol/L抑制作用最为明显(P<0.01).ghrelin在6～24 h内均能明显抑制HASMC增殖,在24 h达最高峰(P<0.01).10-6 mol/L ghrelin能明显上调Mfn-2 mRNA和蛋白的表达(P<0.01).10-6 mol/L ghrelin在18 h上调Mfn-2 mRNA和蛋白表达的作用最为明显(P<0.01).结论 ghrelin可能通过上调Mfn-2的表达来抑制HASMC增殖.     

脂质体介导10-23脱氧核酶对人血管平滑肌细胞增殖的影响

目的 :研究针对增殖细胞核抗原 (PCNA)基因特异性的 10 - 2 3脱氧核酶 (DNAzyme)对人血管平滑肌细胞(VSMC)PCNA的表达和细胞增殖的影响。方法 :应用脂质体转染法将不同寡聚核苷酸转入体外培养的人脐动脉VSMC。采用3 H -TdR掺入法观察空白对照组、脂质体组和等摩尔浓度 (1.0 μmol/L)的DNAzyme、反义寡核苷酸 (A SODN)、mDNAzyme在干预 2d后VSMC的DNA合成速率 ;免疫组化检测 1.0 μmol/L的DNAzyme和ASODN转染 2d后对PCNA表达的影响 ;四甲基偶氮唑蓝 (MTT)比色法分析不同剂量的DNAzyme(0 .12 5、0 .2 5、0 .5、1.0、2 .0 μmol/L)转染5d后VSMC增殖活性。结果 :1.0 μmol/LDNAzyme和ASODN转染 2d后 ,VSMC的3 H -TdR掺入量均较对照组显著减低 (P <0 .0 5 ) ,DNAzyme较ASODN更显著减低 (P <0 .0 5 ) ;1.0 μmol/LDNAzyme作用 2d后细胞增殖抑制率为 6 4 .5 % ,高于ASODN组 (37.7% )、mDNAzyme组 (11.9% )和脂质体组 (3.3% )。DNAzyme和ASODN处理细胞 2d后 ,PCNA的表达水平 (平均光密度值 )分别为 0 .2 95 6± 0 .0 2 5 1和 0 .3880± 0 .0 92 5 ,明显低于空白对照组 0 .7342± 0 .2 136 (P <0 .0 1)。转染 5d后 ,DNAzyme的作用呈剂量依赖性 ,0 .12 5 μmol/LDNAzyme处理后VSMC的增殖较对照组低 (P <0 .0 5 )。     

Establishment and pathological study of models of chronic ob structive pulmonary disease by SO2 inhalation method

Objective To establish rat models of chronic obstructive pulmonary disease (COPD) and study the pathological characteristics of airflow obstruction.
Method SO₂ inhalation method was used to establish rat models. After exposure to SO₂ for 7 weeks, peak expiratory flow (PEF), peak inspiratory flow (PIF), intratracheal pressure (IP) and IP slope in rat were measured by Maclab data recording and analysis system. Experimental rats with PEF less than 80% of the mean of the normal rats were classified as airflow obstructed, while those with PEF greater than 80% of mean of normal rats were non-obstructed. Pathological changes in airway and lung tissue were compared between these two groups.
Result In experimental animals, PEF was significantly decreased (P<0.005) and IP slope increased (P<0.001) as compared with normal rats. Epithelial damage, goblet cell hyperplasia and inflammatory cell infiltration in cartilaginous bronchi were more remarkable in experimental rats with airflow obstruction than those without airflow obstruction (P<0.001, <0 .01, <0.001, respectively). Furthermore, pathological changes in airway lumen, epithelium and airway wall in membranous and respiratory bronchioles were more marked in experimental rats with airflow obstruction than those without airflow obstruction (P<0.001 or P<0.05). There was a negative correlation between PEF values and epithelial hyperplasia, goblet cell hyperplasia, inflammatory cell infiltration, smooth muscle hyperplasia and mucous plug in membranous and respiratory bronchioles (P<0.001 or P<0. 05).
Conclusion SO₂ inhalation may cause chronic bronchitis with airflow obstruction, i.e. COPD in rats. COPD was induced in 64% (16 of 25) of the experimental group rats.     

钙调神经磷酸酶信号通路在AngⅡ刺激的大鼠VSMCs增殖中的作用

目的:探讨钙调神经磷酸酶（calcineurin,CaN）依赖的信号通路在血管紧张素Ⅱ（Ang Ⅱ）刺激的大鼠血管平滑肌细胞（VSMCs）增殖中的作用。方法:采用组织贴块法,体外原代培养大鼠胸主动脉平滑肌细胞;Ang Ⅱ刺激培养的大鼠 VSMCs增殖;CaN特异性抑制剂环孢素 A（CsA）阻断Ang Ⅱ刺激的大鼠VSMCs中CaN依赖的信号转导通路;观察各因素对CaN活性、细胞增殖活度、增殖细胞核抗原（PCNA）表达水平的影响。结果:在一定范围内,Ang Ⅱ（10-8～10-6mol/L）以浓度依赖性方式增加VSMCs的CaN活性,同时细胞增殖活度（AMTT值）增高,与对照组相比,差异有统计学意义（P<0.01或P<0.001）。CsA（1 μmol/L）抑制CaN活性后,明显降低Ang Ⅱ（10-7mol/L）刺激的VSMCs增殖活度及核内PCNA的表达水平（OD值）,与Ang Ⅱ组比较,差异显著（P<0.001）。结论:CaN依赖的信号通路在Ang Ⅱ刺激的血管平滑肌细胞增殖中发挥重要作用。