胰岛素对大鼠血管平滑肌细胞表型转化的影响

目的:探讨胰岛素对体外培养的血管平滑肌细胞(vascular smooth muscle cell,VSMC)表型转化的影响.方法:用酶消化法分离大鼠VSMC,采用免疫组织化学法检测胰岛素作用后VSMC肌动蛋白(α-SM actin),RT-PCR 检测胰岛素作用后VSMC中bFGF、TGF-β、PDGF、matrix Gla和OPN各目的基因mRNA相对表达水平.结果:胰岛素组VSMC的3H-TdR掺入值比对照组升高47％(P<0.01), VSMC α- SM actin免疫组化结果显示对照组的α-SM actin比胰岛素组染色深.而matrix Gla和OPN在培养的VSMC中mRNA的表达量,胰岛素组明显高于对照组(P<0.05),同时bFGF、TGF-β、PDGF的表达胰岛素组也明显高于对照组(P<0.05).并可明显见到细胞骨架F-actin、G-actin重新分布.结论:在合成表型的matrix Gla和OPN表达量明显高于收缩表型,而合成表型的α- SM actin表达量明显低于收缩表型.提示胰岛素对VSMC的表型转化起了一定作用.胰岛素在促进了VSMC增殖的同时,伴有VSMC由收缩型转变为合成型及细胞骨架F-actin、G-actin重新分布.     

ORC1基因RNA干扰对大鼠血管平滑肌细胞表型的影响

目的 以RNA干扰抑制血管平滑肌细胞(vascular smooth muscle cells,VSMCs)ORC1基因,探讨ORCl基因表达抑制后对VSMCs表型的影响.方法 实验设置正常对照组、阴性siRNA组及阳性siRNA组.应用Western blot检测ORC1基因表达的变化;免疫荧光染色后激光共聚焦显微镜观察细胞形态结构;应用流式细胞仪检测细胞表型标志蛋白表达.结果 ①siRNA转染后,阳性siRNA组ORCI基因表达水显著降低,而正常对照组及阴性siRNA组间ORC1基因表达水平无显著差异.②siRNA转染后,阳性转染组细胞呈现收缩型形态特征,空白对照组及阴性对照组细胞呈现合成型形态特征.③siRNA转染使ORCI表达减弱后,VSMCs收缩型标志物α-平滑肌肌动蛋白(α-SM actin)和平滑肌肌动蛋白重链2(SM-2)表达水平明显升高.合成型标志物骨桥蛋白(osteopontin)表达水平明显降低,空白对照组及阴性对照组间3种标志物表达水平无显著差异.结论 RNA干扰介导的ORC1基凶沉寂可促使VSMCs由合成型向收缩型转化.     

PDGF-AB对成纤维细胞表达α-SM actin的影响

目的:研究PDGF-AB对成纤维细胞表达α-SM actin的影响,探讨PDGF-AB促进伤口愈合的作用机制. 方法:取体外培养的人正常皮肤成纤维细胞,用免疫组化(SABC)观察PDGF-AB对成纤维细胞表达α-SM actin的影响. 结果:PDGF-AB能诱导成纤维细胞表达α-SM actin,且呈剂量依赖性关系. 结论:PDGF-AB可能通过刺激成纤维细胞表达α-SM actin,促进成纤维细胞向肌成纤维细胞转化,增加伤口收缩,促进伤口愈合.     

转化生长因子β1和α平滑肌肌动蛋白在肝内胆管癌血管形成中的作用

目的 探讨转化生长因子β1(transforming growth factorβ1,TGF-β1)和 α 平滑肌肌动蛋白(αsmooth muscle actin,α-SMA)在肝内胆管癌血管形成中的作用.方法 选择50例肝内胆管癌患者的癌组织和远端正常肝组织,通过HE染色观察基本病理变化,天狼猩红染色观察胶原沉积情...     

α—平滑肌肌动蛋白在掌腱膜挛缩症中的表达及意义

为了解国人掌腱膜挛缩症的病理特点,采用免疫组化染色辅以计算机定量分析技术,对17例病变和9例正常掌腱膜组织中α—平滑肌肌动蛋白(简称α—SM actin)的分布和含量进行检测。结果显示:病变组掌腱膜组织中广泛分布有α—SM actin,其含量明显高于正常组(P<0.001)。提示α—SM actin在掌腱膜挛缩症中起重要作用。     

α受体介导的雌二醇对大鼠结肠平滑肌细胞RhoA/ROCK通路的影响

目的：探讨17β?雌二醇（17β?estraial,E2）对大鼠结肠平滑肌细胞（smooth muscle cell,SMC）RhoA/ROCK通路的影响。方法：体外分离培养Sprague?Dawley（SD）大鼠结肠SMCs。细胞免疫荧光双标法观察ROCK1、雌激素受体（ER)α与α肌动蛋白（α?SMA）共表达。不同浓度和不同时间E2刺激后,以反转录实时荧光定量qRT?PCR、Western blot检测RhoA、ROCK1表达。不同浓度ROCK抑制剂Y27632刺激后,Western blot检测E2对其下游蛋白表达情况。观察ER阻断剂ICI182780、牛血清白蛋白结合的E2（BSA?E2）、ERα激动剂PPT及ERβ激动剂DPN在E2影响RhoA、ROCK1表达的作用。构建ERα、ERβ的siRNA,观察E2刺激对转染后SMC中RhoA、ROCK1表达的影响。结果：细胞免疫荧光示结肠SMC上 ROCK1与α?SMA共表达。50 nmol/L E2刺激24 h可显著抑制大鼠结肠SMC上RhoA、ROCK1表达（P < 0.05）。Y27632可浓度依赖性抑制下游蛋白p?MYPT1、p?CPI17、p?MLC表达。E2组及PPT组RhoA、ROCK1表达显著低于其他各组（P < 0.05）。ERα siRNA组可上调RhoA、ROCK1表达（P < 0.05）。结论：大鼠结肠SMC上存在ROCK1,E2可抑制RhoA/ROCK通路表达,该作用由雌激素α受体介导。     

丹参酚酸B盐对转化生长因子β1活化的大鼠肝星状细胞内细胞外信号调节激酶信号转导通路的抑制作用

目的:探讨丹参酚酸B盐(salvianolic acid B,SA-B)对转化生长因子β1 (transforming growth factor-β1,TGF-31)活化的大鼠肝星状细胞(hepatic stellate cell,HSC)内细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)信号转导通路的影响.方法:采用原位灌流酶消化及Nycodenz密度梯度离心的方法分离正常大鼠HSC,将TGF-β1和SA-B直接添加于原代HSC的无血清培养液中.蛋白印记法检测HSC内磷酸化和总ERK,以及有丝分裂原激活蛋白激酶激酶MEK、有丝分裂原活化蛋白激酶激酶激酶Raf、α平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和Ⅰ型胶原蛋白的表达.结果:SA-B可抑制正常原代HSC及TGF-β1刺激的HSC内ERK上游激酶MEK的磷酸化;而对正常原代HSC及TGF-β1刺激的HSC内MEK上游激酶Raf-1的磷酸化无明显抑制.SA-B对TGF-β1刺激的HSC内α-SMA表达有明显的抑制作用,SA-B与ERK阻断剂联用,几乎接近完全阻断α-SMA的表达.SA-B对正常培养的HSC和经TGF-β1刺激的HSC Ⅰ型胶原蛋白的合成都有明显的抑制作用.结论:SA-B抑制TGF-β1刺激的HSC内ERK信号转导通路的作用环节在于抑制了MEK的磷酸化.SA-B通过抑制TGF-β1的ERK信号转导通路,减少了因HSC活化导致的α-SMA表达和Ⅰ型胶原蛋白合成增加.     

动脉粥样硬化时动脉平滑肌细胞增殖及表型转变的若干分子调控机制

动脉粥样硬化(Atherosclerosis,AS)是一种以细胞增生为主要病变的疾病,近年来,众多的研究表明动脉平滑肌细胞(Smooth muscle cell,SMC)是动脉粥样斑块中最主要的细胞成份,并且动脉平滑肌细胞至少有两种细胞表型,即“合成型”(Syn-thetic Phenotype)和“收缩型”(ContractilePhenotype)。在一定因素刺激下,细胞发生分化增殖,表型发生改变,分泌大量细胞外基质,可以引起细胞内外脂质沉积,形成动     

TGF-β1对大鼠腹膜间皮细胞转分化的影响及其机制

目的:研究转化生长因子(transforming growth factor β1,TGF-β1)对大鼠腹膜间皮细胞(rat peritoneal mesothelial cells,RPMCs)转分化的影响,并探讨其可能的机制.方法:体外培养SD大鼠原代腹膜间皮细胞,静止24 h后,随机分为正常对照组和TGF-β1(10 μg/L)刺激组.用TGF-β1(10 μg/L)刺激RPMCs不同时间,分别用RT-PCR方法和Western免疫印迹法检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、E-钙黏素、Ⅰ型胶原的mRNA和蛋白的表达.用Western免疫印迹法检测总RhoA的蛋白表达水平,活化的RhoA由膜蛋白提取试剂盒提取,并用Western免疫印迹法评估.结果:TGF-β1刺激RPMCs能导致E-钙黏素mRNA和蛋白表达下调;α-SMA,Ⅰ型胶原mRNA和蛋白表达上调,同时RhoA蛋白表达上调,呈时间依赖性.结论:TGF-β1诱导了大鼠腹膜间皮细胞转分化,其机制可能通过RhoA介导的信号通路.     

月舒滴丸对大鼠子宫内膜及平滑肌细胞OTR表达的影响

目的:探讨月舒滴丸对大鼠子宫平滑肌细胞缩宫素受体(Oxytocin receptor,OTR)基因表达及对大鼠子宫内膜上皮细胞前列腺素(Prostaglandin,PG)合成的影响,阐明月舒滴丸治疗原发性痛经的作用机制。方法:分别采用雌二醇(Estradiol,E2)和缩宫素(Oxytocin,OT)刺激,复制子宫平滑肌细胞OTR高表达模型,采用RTPCR法,探讨月舒滴丸对子宫平滑肌细胞OTR基因表达的影响;采用Elisa法观察月舒滴丸干预对大鼠子宫内膜上皮细胞PGF2α、PGE2含量变化的影响。结果:月舒滴丸能明显降低大鼠子宫平滑肌细胞OTR基因表达,并可以显著降低大鼠子宫内膜上皮细胞PGF2α水平,提高PGE2表达,降低子宫内膜细胞PGF2α/PGE2水平。结论:通过月舒滴丸的干预,可减轻缩宫素的刺激,从而使子宫平滑肌细胞OTR低表达,减少子宫平滑肌细胞对OT的敏感性,并且能够有效调节大鼠子宫内膜上皮细胞前列腺素水平。     

Increased apoptosis and decreased density of medial smooth muscle cells in human abdominal aortic aneurysms

Objective To determine the increase of apoptosis and the decrease of smooth muscle cells (SMCs) density in human abdominal aortic aneurysms (AAA).Methods In situ terminal transferase-mediated dUTP nick end labeling (TUNEL) was employed to detect apoptosis of SMCs in patients with AAA (n=25) and normal abdominal aortae (n=10). Positive cells were identified by specific cell marker in combination with immunohistochemistry. Meanwhile SMC counting was performed by anti-α-actin immunohistostaining to compare the SMC density. Results TUNEL staining revealed that there was significantly increased apoptosis in AAAs (average 8.6%) compared with normal abdominal aortae (average 0.95%, P&lt;0.01). Double staining showed that most of these cells were SMCs. Counting of α-actin positive SMCs revealed that medial SMC density of AAAs (37.5±7.6 SMCs /HPF) was reduced by 79.1% in comparison with that of normal abdominal aortae (179.2±16.1 SMCs /HPF, P&lt;0.01). Conclusions Significantly increased SMCs of AAA bear apoptotic markers initiating cell death. Elevated apoptosis may result in a decreased density of SMCs in AAA, which may profoundly influence the development of AAA.     

血管钙化大鼠肾动脉BMP2/Smad1/Runx2信号通路的表达

目的 观察血管钙化大鼠模型肾动脉上骨形态蛋白2（BMP2）/Smad1/Runt相关转录因子2（Runx2）信号通路的表达及其变化规律,探讨BMP2/Smad1/Runx2信号通路激活在肾动脉钙化中的作用。方法 将24只雄性SD大鼠随机分为对照组和钙化组,钙化组建立维生素D3和尼古丁诱导的大鼠血管钙化模型,对照组给予生理盐水和花生油。6周后采用Von Kossa染色检测肾动脉钙化程度,钙离子试剂盒测定大鼠肾动脉钙含量,实时荧光定量PCR检测肾动脉组织中BMP2、Smad1、Runx2 mRNA水平,免疫组化法观察BMP2、Smad1、Runx2及α-平滑肌肌动蛋白（α-SMA）的表达。结果 Von Kossa染色见钙化组大鼠肾动脉有大量黑色颗粒沉淀。钙化组血管钙含量高于对照组（ P<0.05)。与对照组相比,钙化组大鼠肾动脉组织BMP2、Smad1、Runx2 mRNA的水平升高（P<0.05),BMP2/Smad1/Runx2信号通路蛋白在肾动脉的表达亦同步上调,而α-SMA的表达较对照组下降（P<0.05)。相关分析表明,大鼠肾血管钙含量与肾动脉组织BMP2 mRNA（r =0.655, P<0.05)、Smad1 mRNA (r =0.735, P<0.05)、Runx2 mRNA ( r=0.734, P<0.05)均呈正相关。结论 BMP2/Smad1/Runx2通路的表达变化与肾动脉钙化的严重程度相关,提示BMP2/Smad1/Runx2信号通路参与了肾动脉钙化的发生发展。     

Effects of calcium-activated chloride channels on proliferation of pulmonary artery smooth muscle cells in rats under chronic hypoxic condition

Objective: To investigate the effects of calcium-activated chloride (C1Ca) channels on proliferation of pulmonary artery smooth muscle cells(PASMCs) in rats under chronic hypoxic condition. Methods:The cultured PASMCs were placed under normoxic and chronic hypoxic conditions:The cells were observed by light and electron microscope; The cell cycles were observed by flow-cytometry; Immunocytochemistry staining was used to detect the expressions of PCNA, c-los and c-jun of PASMCs; Cytoplasmic free Ca2 con-centration ([Ca2 ]) in PASMCs was investigated by fluorescent quantitation using fluorospectrophotometer. Results:The PASMCs were contractile phenotype under normoxic conditions. Observation by transmission electron microscope: In kytoplasm of contractile phenotype cells, myofilament bundles were abundant and the content of cell organs such as Golgi's bodies were rare. The PASMCs were synthetic phenotype under chronic hypoxic condition. There were inereased free ribosomes, dilated rough endoplasmic reticulums, highly developed Golgi complexes, decreased or disappeared thick filaments and dense body in kytoplasm of synthetic phenotype cells. After NFA and IAA-94, the situations were reversed The number of S4,GzM PASMCs were significantly increased in chronic hypoxic condition; The NFA and IAA-94 were shown to significantly decrease them from (28.6±1.0)% to (16.0±1.6)% and the number of G0G1 PASMCs significantly increased from (71.4 ±1.9)% to (83.9±1.6)% (P＜ 0.01). In chronic hypoxic conditions, the expression of proliferating cell nucleus antigen was significantly increased; The NFA and IAA-94 were shown to significantly decrease it from (81±6)% to (27±7)%(P＜0.01). The expression of c-los and c-jun were significantly increased in-chronic hypoxic conditions; The NFA and IAA-94 were shown to significantly decrease them from 0.15 ± 0.02, 0.32 ± 0.05 to 0.05 ± 0.01, 0.12±0.05, respectively (P＜ 0.01); Under chronic hypoxic conditions, [Ca2 ]. Was increased; The NFA and IAA-94 decreased it from (281.8±16.5)nmol/L to (117.7±15.4)nmol/L(P＜0.01). Conclusion:Hypoxia initiated the change of PASMCs from contractile to synthetic phenotype and increased proliferation of PASMCs. NFA and IAA-94 depressed cell proliferation by blocking C1Ca channels in hypoxic condition. These may play an important role in proliferation of PASMCs under chronic hypoxic conditions.     

Early Results of Coronary Endarterectomy Combined with Coronary Artery Bypass Grafting in Patients with Diffused Coronary Artery Disease

Background:

It is still a challenge for the cardiac surgeons to achieve adequate revascularization for diffused coronary artery disease (CAD). Coronary endarterectomy (CE) offers an alternative choice of coronary artery reconstruction and revascularization. In this study, short-term result of CE combined with coronary artery bypass graft (CABG) was discussed in the treatment for the diffused CAD.

Methods:

From January 2012 to April 2014, 221 cases of CABG were performed by the same surgeon in our unit. Among these cases, 38 cases of CE + CABG were performed, which was about 17.2% (38/221) of the cohort. All these patients were divided into two groups: CE + CABG group (Group A) and CABG alone group (Group B). All clinical data were compared between the two groups, and postoperative complications and in-hospital mortality were analyzed. The categorical and continuous variables were analyzed by Chi-square test and Student''s t-test respectively.

Results:

Diabetes mellitus, hypertension, hyperlipidemia, and peripheral vascular disease were more common in group A. In this cohort, a total of 50 vessels were endarterectomized. Among them, CE was performed on left anterior descending artery in 11 cases, on right coronary artery in 29 cases, on diagonal artery in 3 cases, on intermediate artery in 2 cases, on obtuse marginal artery in 5 cases. There was no hospital mortality in both groups. The intro-aortic balloon pump was required in 3 cases in Group A (3/38), which was more often than that in Group B (3/183). At the time of follow-up, coronary computed tomography angiogram showed all the grafts with CE were patent (50/50). There is no cardio-related mortality in both groups. All these patients were free from coronary re-intervention.

Conclusions:

Coronary endarterectomy + CABG can offer satisfactory result for patients with diffused CAD in a short-term after the operation.     

子宫动脉栓塞治疗子宫肌瘤的临床研究

目的：探讨子宫动脉栓塞术治疗子宫肌瘤的疗效。方法：选择25例子宫肌瘤患,采用Seldinser技术经一侧股动脉穿刺,将导管超声选择性插入双侧子宫动脉远端,经导管注入PVA和明胶海绵微粒,阻断子宫肌瘤的血供。结果：血管造影显示子宫肌瘤血供丰富,均为双侧子宫动脉供血。栓塞后血管造影见子宫肌瘤血供完全阻断,技术成功率为100％。随访结果表明,患临床症状缓解消失,肌瘤和子宫体积明显缩小。术后有不同程度的盆腔疼痛、低热,经对症处理后症状逐渐消失。结论：子宫动脉栓塞术治疗子宫肌瘤是一种疗效显的微创治疗新方法。     

海藻酸钠微球子宫动脉栓塞治疗子宫肌瘤

目的评价海藻酸钠微球在子宫肌瘤动脉栓塞治疗中的应用价值。材料与方法3例多发性子宫肌瘤患者施行了超选择性子宫动脉栓塞术。所用栓塞剂为直径500μm-700μm海藻酸钠微球。对栓塞前后子宫肌瘤的大小、数量、临床症状及体征,以及卵巢激素水平进行比较。术后1个月、3个月进行B超随访,术后3个月进行卵巢激素测定。以后每隔1年B超随访。结果3例多发子宫肌瘤患者,年龄40岁、41岁、43岁（平均41.5岁）。治疗前与治疗后三个月卵巢激素水平无明显变化。栓塞后1个月时症状明显好转,肌瘤缩小平均40%,其中4个小肌瘤消失。术后三个月肌瘤缩小50～60%,有5个小肌瘤消失。临床症状完全消失。平均每例随访42个月,未出现新的肌瘤,残留肌瘤体积随时间延长,逐渐变小。栓塞不良反应主要为疼痛、恶心、呕吐及发热。无严重不良反应发生。结论海藻酸钠微球用于子宫肌瘤动脉栓塞治疗方便、安全、疗效确切。     

慢性低氧钙激活氯通道在大鼠肺动脉平滑肌细胞增殖中的作用

目的探讨慢性低氧时钙激活氯通道（ClCa）在大鼠肺动脉平滑肌细胞（PASMCs）增殖中的作用。方法将PASMCs分别置于常氧及慢性低氧下,采用形态学、流式细胞术、免疫细胞化学法,观察ClCa阻滞剂尼氟灭酸（NFA）和indaryloxyacetic acid（IAA-94）对PASMCs增殖的影响。结果慢性低氧：①PASMCs呈合成表型,而常氧为收缩表型,NFA和IAA-94干预后呈合成表型的PASMCs向收缩型转变;②S＋G2M期细胞所占比例增高,NFA和IAA-94干预后减低,且G0G1期细胞所占比例增加（P〈0.01）;③PCNA表达阳性率增高,NFA和IAA-94干预后降低（P〈0.01）;④c-fos和c-jun蛋白阳性染色A增高,NFA和IAA-94干预后降低（P〈0.01）。结论慢性低氧引起细胞表型改变,可促进细胞增殖,而抑制ClCa的活性可抑制细胞的增殖,提示ClCa参与了低氧PASMCs的增殖。     

64排螺旋CT冠状动脉成像诊断心肌桥和壁冠状动脉的临床应用

目的:探讨64排螺旋CT冠状动脉成像诊断心肌桥-壁冠状动脉(MB-MCA)的检查方法及临床应用。方法:回顾性分析330例患者64排CT冠状动脉成像资料。结果:62例有心肌桥-壁冠状动脉(MB-MCA)存在,其中49例发生在左前降支及其分支,11例发生在左回旋支及其分支,2例发生在右冠状动脉。结论:64排螺旋CT冠状动脉成像能较好地显示壁冠状动脉与心肌桥的关系,有助于壁冠状动脉长度和心肌桥厚度的评价,为临床诊断和治疗提供了更有价值的信息。     

CT冠状动脉血管成像判定心肌桥-壁冠状动脉形态学特征及其与冠状动脉粥样硬化病变关系

目的探讨采用CT冠状动脉血管成像观察心肌桥-壁冠状动脉的形态学特征,并分析其与冠状动脉粥样硬化病变的关系。方法选取100例经128层螺旋CT冠状动脉成像技术证实的单纯深在型心肌桥-壁冠状动脉患者(深在型)、100例心肌桥近端冠状动脉合并粥样硬化患者(硬化组)的CT冠状动脉成像数据进行统计分析,比较两组患者的影像学指标差异,并采用Logistic回归分析心肌桥-壁冠状动脉的形态学特征与冠脉粥样硬化的关系。结果深在型组和硬化组的心肌桥厚度、壁冠状动脉近端成角率差异无统计学意义(P0.05),两组间壁冠状动脉长度、壁冠状动脉两端平滑率、壁冠状动脉两端成角率、壁冠状动脉远端成角率差异均具有统计学意义(P0.05);壁近段冠状动脉粥样硬化的危险因素为年龄(OR=1.872)、两组间壁冠状动脉长度(OR=2.031)、壁冠状动脉两端成角(OR=1.376)、壁冠状动脉远端成角(OR=1.448),壁冠状动脉两端平滑是保护因素(OR=0.471)。结论心肌桥-壁冠状动脉复合体的形成与冠脉近段粥样硬化形成密切相关。     

64层螺旋CT冠状动脉血管成像的临床应用价值

目的评价64层螺旋cT用于诊断冠状动脉病变的临床价值。方法搜集35例患者64层螺旋cT冠状动脉血管成像的完整资料,并以近期实施的选择性冠状动脉造影（SCA）结果为金标准进行对比,以评价64层螺旋CT冠状动脉成像的应用价值。结果35例病例可用于评估的冠状动脉共计356个节段,成功率91．28％。其中SCA共显示73个节段冠脉有中、重度狭窄（≥50％）,其结果与冠状动脉成像结果比较,对病变诊断的敏感性为90．7％,特异性为95．3％,准确性为94．7％,阳性预测值为92．9％,阴性预测值为97．6％;支架术后7例,共11段血管段上有支架;搭桥术后2例,共4支桥血管。结论64层螺旋cT冠状动脉血管成像作为一种无创的检查手段,在诊断冠状动脉病变中有较高的敏感性和特异性,适用于冠心病的早期筛查及冠状动脉搭桥和支架置入术前评价和术后复查的首选方法。