超声引导下凝血酶注射治疗假性动脉瘤的价值

目的：探讨在经皮超声引导下注射凝血酶（UGTI）治疗假性动脉瘤的应用价值。方法：对于27例介入术后股动脉假性动脉瘤（PsA）,采用彩超引导下假腔内注射凝血酶致栓塞治疗,观察疗效,血栓形成时间和凝血酶的用量。并在治疗后10min、1d、5d、15d复查彩超。结果：瘤内血栓形成迅速,时间为2～4min,凝血酶用量为（350±150）IU（250IU／m1）;治愈率100％。患者均系一次治疗成功,无过敏反应等并发症发生。27例患者术后随访,均无复发。结论：超声引导下凝血酶注射治疗具有创伤小,操作简便,疗效确切的优点,可作为经股动脉介入治疗术后形成的假性动脉瘤的首选治疗方法。     

Effects of thrombin on intracellular calcium and pH in human and murine platelets

The aim of this study was to compare the effect of thrombin (Thr) on cytosolic calcium [Ca 2 ] + i and intracellular pH [pH]i in human and murine platelets. Rich-platelet suspensions from both species were loaded with Fura-2 (2 w M) or BCECF (0.75 w M) by incubation with their respective acetoxymethyl esters to measure cytosolic calcium [Ca 2+ ] i or intracellular pH [pH]i, respectively. Suspensions were challenged with increasing concentrations of Thr, from 0.1 to 10 IU/ml. Basal [Ca 2+ ] i in human platelets was 98 - 6 and 99.1 - 9 nM in rat platelets ( n =20). Thr increased \[Ca2+] i , EC 50 was 1.1 - 0.04 in human and 0.97 - 0.06 IU/ml in rat platelets ( n =7). Extracellular Mg 2+ (4 or 8 mM) abolished Thr response on [Ca 2+ ] i . [pH]i in human was 7.09 - 0.08 and 7.11 - 0.04 in rat platelets. Thr induced alkalinization of platelets in both species. Our results indicate that the potency of Thr to change [Ca 2+ ] i and [pH]i was similar in both species, allowing for comparisons between human and murine platelets and to extrapolate results from an animal model to human pathology.     

Inhibitory effect of glyburide on thrombin-induced platelet aggregation and phosphoinositide metabolism in normal human platelets

We previously reported the effects of diet, sulphonylureas or insulin on thrombin-induced platelet aggregation, phosphoinositide metabolism and protein phosphorylation in non-insulin-dependent diabetes mellitus (NIDDM) patients. To clarify the mechanism of glyburide and insulin on platelet function, here we studied the in vitro effects of glyburide and insulin on thrombin-induced metabolic changes using normal human platelets. Platelet aggregation stimulated with <0.5 U/ml thrombin, 0.75-3 mu M adenosine diphosphate (ADP) or 1 mu g/ml collagen was significantly lower in glyburide-treated platelets, but not in insulin-treated platelets, than in untreated ones (control). Thrombin-induced incorporation of 32P radioactivity into phosphatidic acid (PA) in glyburide-treated platelets was lower than that in control but not in insulin-treated platelets. Phosphorylated proteins of platelets induced by thrombin and 12- O -tetradecanoylphorbol 13-acetate (TPA) in glyburide-treated platelets were suppressed, but not in insulin-treated platelets, compared with control. These results suggest that glyburide induces suppression of thrombin-induced activation of phospholipase C, which mediates hydrolysis of PIP and PIP2 and production of PA, and subsequently inhibits platelet aggregation.     

脑出血后凝血酶与脑水肿关系及水蛭粉的疗效

目的　探讨凝血酶在脑出血后脑水肿形成机制中的作用以及水蛭粉治疗脑水肿的疗效。方法　分别制作大鼠自体全血、凝血酶、肝素抗凝血、生理盐水及假手术脑血肿模型 ,观察不同时间血肿周围脑组织含水量 ,Na+ 、K+ 含量的变化以及病理改变 ;并观察水蛭粉 (2 5 0mg/kg口服 ,每天 2次 ,治疗 5d)治疗脑水肿的疗效。结果　大鼠脑内注射全血或凝血酶后 ,脑组织的含水量、电解质含量的变化相一致 ;在 2 4h时 ,脑水含量、Na+ 含量较其它 3组高 ,而K+ 含量则较低 (P <0 0 5 )。水蛭粉治疗后脑水含量、Na+ 、Ca2 + 含量较对照组降低 ,而K+ 含量升高 (P <0 0 5 )。结论　凝血酶可能在脑出血后脑水肿形成机制中起重要作用 ;水蛭粉对大鼠全血和凝血酶导致脑水肿有显著影响。     

凝血酶通过内皮素介导刺激肾小球系膜细胞增生

研究内皮素－１在凝血酶刺激有肾小球系膜细胞增生中的作用。方法在培养肾小球系膜细胞中进行试验。（１）ＨＭＣ在凝血酶刺激下增生,呈剂量依赖并与刺激时间有关,当凝血酶１６Ｕ／ｍｌ刺激１６小时时ＨＭＣ增生达高峰;     

不同损伤时期血管内膜的增殖和凝血酶的影响

目的：探讨内皮拉伤后血管内膜增殖反应的动态变化,和凝血酶对损伤血管增殖反应的影响。　　方法：Ｆｏｇａｒｔｙ 导管拉伤大鼠主动脉,不同时期处死,病理观察血管内膜厚度,免疫组织化学观察增殖细胞核抗原在损伤血管的表达;并将离体的血管环在体外加入不同浓度的凝血酶［０ Ｕ／ｍ ｌ（对照组）、１ Ｕ／ｍ ｌ（１ Ｕ凝血酶组）和５ Ｕ／ｍ ｌ（５ Ｕ凝血酶组）］培养１２小时,测定单位重量血管环氚标记胸腺嘧啶脱氧核苷（３Ｈ－ＴｄＲ）的掺入量。　　结果：拉伤后血管内膜／中膜面积值于４～６周达到高峰,７～８周增厚的内膜几乎完全消退。增殖细胞核抗原的表达与血管内膜增殖反应基本一致。３Ｈ－ＴｄＲ掺入于术后３日和３ 周出现２个高峰。加入凝血酶１ Ｕ／ｍ ｌ或５ Ｕ／ｍ ｌ,３日的３Ｈ－ＴｄＲ掺入量增加,与０ 日比较有显著性差异Ｐ＜ ０．０５,４周后３Ｈ－ＴｄＲ掺入进入低反应期。　　结论：血管损伤后３ 日是细胞分裂最为活跃的时期,对凝血酶促增殖作用也最敏感,４周后为低反应期,因此及早有效抗凝对于控制血管增生和再狭窄有十分重要的意义。     

急性冠脉综合征患者pentraxin-3与凝血酶活化的纤溶抑制物的变化

目的了解急性冠脉综合征(Acute Coronary Syndrome,ACS)患者炎性指标pentraxin-3与纤溶指标凝血酶活化的纤溶抑制物(Thrombin activatable finolysis inhibitor,TAFI)的变化。方法检测比较102例急性心肌梗死(AMI组)、81例不稳定心绞痛(UAP组)及23例健康体检者(对照组)的血浆pentraxin-3、TAFI水平。结果急性冠脉综合征患者血浆pentraxin-3、TAFI明显高于对照组,差异有统计学意义(P<0.01);AMI组血浆pentraxin-3及TAFI水平高于UAP组,差异有统计学意义(P<0.05)。pentraxin-3浓度与TAFI浓度呈正相关(r=0.17,P<0.05)。结论 pentraxin-3与TAFI共同在ACS的发生发展中起重要作用。     

凝血酶对血脑屏障通透性影响及其机制的实验研究

目的　探讨凝血酶 (TB)对血脑屏障通透性的影响及其可能机制。　方法　将TB或TB +组织蛋白酶G(CT G)立体定向注入SD大鼠右侧基底节 ,测定其血脑屏障通透性和脑水含量 ;将脑微血管内皮培养液中加入TB或TB +CT G ,相差显微镜动态观察内皮细胞形态的变化 ,免疫组织化学技术检测基质金属蛋白酶 2 (MMP 2 )表达的改变。　结果　TB脑内注射后 6h ,同侧基底节区血脑屏障通透性为 (0 4 19± 0 0 16 )A/mg (P <0 0 5 ) ,2 4h时血脑屏障通透性为 (0 5 94± 0 0 18)A/mg(P <0 0 1) ,持续至 4 8h (P <0 0 5 )逐渐消退 ,脑组织水含量的变化规律与血脑屏障通透性的变化类似。TB +CT G组在各个时间点血脑屏障通透性和脑组织水含量与对照组比较差异均无显著性 (P >0 0 5 )。TB使内皮细胞发生收缩 ,细胞收缩程度具有时间依赖性。TB入培养液 6h后 ,内皮细胞MMP 2表达水平为 (0 391± 0 0 17)A ,与对照组比较差异有显著性 (P <0 0 1)。TB +CT G加入培养液后 ,细胞形态、MMP 2表达与对照组比较差异均无显著性 (P >0 0 5 )。　结论　TB增加血脑屏障通透性是其诱发脑水肿形成的主要机制之一 ,TB通过激活蛋白酶激活受体 1(PAR 1) ,使内皮细胞发生收缩 ,促进MMP 2表达 ,是TB增加血脑屏障通透性的可能机制     

水蛭素对凝血酶诱导的血管平滑肌细胞增殖的影响

为研究重组水蛭素对凝血酶诱导的免胸主动脉血管平滑肌细胞增殖的影响及其作用机制。选用第2～5代培养的血管平滑肌细胞，在与凝血酶(4．0ku／L)共同孵育的条件下，分别给予水蛭素(6．0ku／L)和肝素(6．0ku／L)进行干预。通过四唑盐比色实验检测细胞生长活性，流式细胞术检测细胞周期，免疫组织化学检测血小板生长因子和增殖细胞核抗原在血管平滑肌细胞中的表达。结果发现，重组水蛭素能明显抑制凝血酶诱导的平滑肌细胞增殖，其抑制作用可能与减低了血管平滑肌细胞对血小板生长因子和增殖细胞核抗原的表达有关。