雷帕霉素与紫杉醇药物洗脱支架对小型猪早期冠状动脉炎症性狭窄治疗的定量冠状动脉造影研究

目的:制作小型猪冠状动脉炎症型支架再狭窄模型,用定量冠状动脉造影、测定支架处血管壁内膜增生程度及炎症因子表达等方法探讨雷帕霉素及紫杉醇涂层支架对炎症性冠状动脉狭窄的抑制作用。方法:小型雄性家猪14头,开胸法将冠状动脉外膜包裹吸附白介素-1β(IL-1β)琼脂糖微粒悬液的纸巾。2周后,用定量冠状动脉造影观察管腔狭窄程度及随机分为雷帕霉素支架组(n=7)、紫杉醇支架组(n=9)及裸支架组(n=8)。1个月后进行随访定量冠状动脉造影测定各组冠状动脉支架节段内和支架内的最小管腔直径(MDL),参照管腔直径(RLD),管腔狭窄百分比(DS),计算出晚期管腔丢失(LLL)进行对比分析。塑料包埋法进行支架血管切片,测定管腔增生面积及血管壁单核细胞趋化因子-1(MCP-1)、肿瘤坏死因子-α(TNF-α)、P-选择素、血管细胞间黏附因子-1(VCAM-1)的表达。结果:冠状动脉外膜包裹IL-1β血管段局限性狭窄平均达到70％以上。裸支架组中冠状动脉支架节段内和支架内的LLL明显高于雷帕霉素支架组和紫杉醇支架组,且再狭窄(DS)率高。雷帕霉素支架组及紫杉醇支架组冠状动脉支架节段内和支架内LLL均低于裸支架组,且明显抑制内膜增生(P<0．01)。但只有雷帕霉素支架组明显下调MCP-1,TNF-α,P-选择素和VCAM-1的表达。结论:雷帕霉素和紫杉醇药物洗脱支架对IL-1β诱发的小型猪冠状动脉炎症性狭窄均有较好的抗内膜增殖作用,但在LLL指标及抑制内膜增生方面雷帕霉素优于紫杉醇药物洗脱支架,有可能是通过直接或间接抗炎机制达到的。     

Rho激酶在小型猪白介素-1β介导的冠状动脉痉挛中的作用机制

目的探讨Rho激酶途径在冠状动脉痉挛发病中的作用机制。方法小型雄性家猪随机分为对照组(n=8)和模型组(n=8),冠状动脉外膜分别包裹吸附含生理盐水和白介素(IL)-1β琼脂糖微粒悬液的纸巾。2周后,采用冠状动脉造影观察管腔狭窄程度及5-羟色胺诱发冠状动脉痉挛情况。测定冠状动脉包裹血管段Rho激酶mRNA表达、肌球蛋白结合亚基磷酸化表达及光镜病理学改变。结果冠状动脉外膜包裹IL-1β血管段发生不同程度的管腔狭窄,5-羟色胺可诱发病变血管段痉挛。模型组与对照组相比,Rho激酶的mRNA表达病变血管段明显上调[(98·20±7·66)%对(63·70±4·26)%,P<0·05];肌球蛋白轻链磷酸酶的肌球蛋白结合亚基磷酸化表达升高(25485±4745对6510±779,P<0·05)。光镜可见病变血管段内膜增殖及炎症细胞聚集现象。结论Rho激酶参与IL-1β介导的冠状动脉痉挛的发生,其可能是通过增加肌球蛋白结合亚基磷酸化水平,抑制肌球蛋白轻链磷酸酶活性及加强钙增敏途径所致。     

雷帕霉素支架对小型猪冠状动脉p27kip表达及内膜增殖的影响

目的　观察小型猪冠状动脉置入支架后p2 7kip表达变化及含 10 0 μg雷帕霉素可降解涂层支架释放的雷帕霉素对p2 7kip表达的影响。方法　球囊 血管以 1 3∶1比例置入过大的裸支架 (n =14 )、单高分子可降解多聚羟基丙酸乙酸 (PLGA)涂层支架 (n =16 )或雷帕霉素支架 (n =16 )并形成冠状动脉损伤模型 ,术后第 1、2、4和 12周时间段处死部分猪。测定 12周时 3组支架血管段的内膜厚度和面积。免疫组化法测定支架置入后 4个时段冠状动脉的p2 7kip表达水平及动态变化。结果　在 12周的随访组 ,裸支架、PLGA支架和雷帕霉素支架的新生内膜平均厚度分别为 (0 38± 0 2 0 )mm、(0 96± 0 5 8)mm和 (0 14± 0 12 )mm(P =0 0 0 4 ) ,3组的新生内膜面积分别为 (3 38± 0 31)mm2 、(6 4 6±2 0 1)mm2 和 (2 0 7± 0 82 )mm2 (P =0 0 0 0 )。置入裸支架 1周后 ,冠状动脉p2 7kip表达处于低水平状态 ,但在第 4周达到最高水平 ,在第 12周时 ,其表达水平恢复至第 1周时水平 ;雷帕霉素支架使整个随访期的p2 7kip表达水平无明显变化 ,均处于高水平表达状态。结论　含 10 0 μg雷帕霉素支架在 12周内有抑制小型猪冠状动脉内膜增殖的明显疗效。裸支架置入后 12周内 ,p2 7kip表达水平在第 4周最高 ;涂层支架释放的     

大鼠血管球囊损伤后平滑肌细胞的增殖与雷帕霉素的干预

目的:研究经皮穿刺血管内成形术后大鼠腹主动脉血管平滑肌细胞增殖程度与p27蛋白水平的关系,及国产雷帕霉素（Rapamyc in）对血管平滑肌细胞增殖的抑制,探讨防治经皮冠状动脉内成形术（PTCA）后再狭窄的有效途径和指标。方法:流式细胞计数法测定p27蛋白、CDK2、cyc lin E在大鼠再狭窄血管平滑肌细胞及对照组的表达水平。结果:雷帕霉素干预后p27蛋白表达水平在PTCA术后各组与对照组相比均有上移,减轻血管狭窄程度（P<0.01）。结论:p27蛋白在增殖的血管平滑肌细胞中表达水平与狭窄程度负相关,国产雷帕霉素可上调p27蛋白水平从而抑制血管狭窄形成。     

通心络抑制白细胞介素1β介导的小型猪冠状动脉早期炎症反应及内膜增殖

目的探讨通心络对白细胞介素1β介导的小型猪冠状动脉早期炎症反应及内膜增殖的抑制作用及可能机制。方法24头小型猪随机均分为假手术组、模型组和通心络组。假手术组和模型组喂养普通饲料,通心络组喂养普通饲料 通心络[1 g/(kg.d)]。2周后麻醉行左侧开胸手术,选择左前降支及回旋支近端管腔外径近似相同的两处节段进行仔细分离,假手术组在血管外膜包裹吸附生理盐水的纸巾,模型组和通心络组包裹吸附白细胞介素1β(2.5μg)的纸巾,术后继续以上喂养方法。2周后采用冠状动脉造影观察各组管腔狭窄情况。造影结束后处死动物,截取包裹血管段,进行病理学检查,并采用逆转录聚合酶链反应,测定各组手术处理血管段单核细胞趋化蛋白1、P选择素、E选择素、细胞间粘附分子1和血管细胞粘附分子1 mRNA的表达。结果冠状动脉造影显示,模型组冠状动脉管腔20%~30%狭窄,病理学检查可见模型组内膜增殖、炎性细胞浸润,平滑肌细胞内膜下迁移。通心络组管腔狭窄程度、内膜增殖及炎性细胞浸润现象较模型组明显减轻。逆转录聚合酶链反应示通心络组单核细胞趋化蛋白1、P选择素、E选择素、细胞间粘附分子1和血管细胞粘附分子1 mRNA表达强度低于模型组(P<0.05)。结论通心络对白细胞介素1β诱导的冠状动脉炎症反应及内膜增殖具有明显的抑制作用。通过抑制细胞因子和粘附因子的表达可能是其主要作用途径。     

聚乳酸聚羟基乙酸共聚物涂层雷帕霉素洗脱支架对小型猪冠状动脉内膜增生的影响

目的:评价生物可降解的聚乳酸聚羟基乙酸共聚物(PLGA)涂层雷帕霉素洗脱支架对健康小型猪冠状动脉内膜增生的影响.方法:26只健康小型猪随机分入316L不锈钢裸金属支架组(316 L组)、L605钴铬合金裸金属支架组(L605组)、PLGA涂层L605支架组(PLGA组)和PLGA涂层雷帕霉素洗脱支架组(雷帕霉素组).各组均包括1周观察终点的猪1只和4周观察终点的猪5只.雷帕霉素组还包括12周观察终点的猪2只.每只猪于左前降支和右冠状动脉各置入同种支架1枚.至观察终点时复查冠状动脉造影并处死取材,通过形态学方法观察内膜增生情况.结果:4周时反映内膜增生的各项指标雷帕霉素组均显著优于其它3组,而其它3组之间没有显著性差异.雷帕霉素组与316L组相比支架上内膜厚度少73%(0.11 mm对0.41 mm),支架间内膜厚度少79%(0.06 mm对0.29 mm),新生内膜面积少69%(0.64 mm2对2.09 mm2),面积狭窄百分比少6l%(18.53%对47.27%).316L组有4例、L605组有3例发生支架内狭窄,而雷帕霉素组无支架内狭窄发生.12周时雷帕霉素组内膜增生有所加重,但较4周时相比除了支架间内膜厚度外其他反映内膜增生的各项指标差异均无统计学意义.结论:采用可降解的PLGA涂层雷帕霉素洗脱支架可以显著抑制健康小型猪冠状动脉支架置入术后4周和12周的内膜增生.     

三氧化二砷洗脱支架诱导猪损伤冠脉平滑肌细胞凋亡

目的观察三氧化二砷（As2O3）多聚左旋乳糖酸（PLLA）涂层支架对猪损伤冠脉平滑肌细胞的凋亡诱导作用及机制。方法在6头小型家猪的冠脉内随机植入裸金属支架、PLLA涂层支架、雷帕霉素洗脱支架和As2O3洗脱支架,7天后处死,TUNEL法检测支架段血管平滑肌细胞凋亡率、免疫组化和Western blot法检测凋亡蛋白p53和增殖细胞核抗原（PCNA）蛋白表达,体外观察As2O3对大鼠血管平滑肌细胞的凋亡诱导作用和p53表达。结果与裸支架和PLLA涂层支架相比,雷帕霉素洗脱支架和As2O3洗脱支架植入局部平滑肌细胞凋亡率以及p53表达明显增多,尤其以As2O3洗脱支架最为明显（P〈0．01）,雷帕霉素洗脱支架和As2O3洗脱支架植入局部PCNA表达较裸支架和PLLA涂层支架明显减少（P〈0．01）,但二者间无显著性差异（P〉0．05）。体外细胞培养显示As2O3呈剂量依赖性的诱导大鼠血管平滑肌细胞凋亡和p53蛋白的表达。结论As2O3洗脱支架具有诱导猪损伤冠脉平滑肌细胞凋亡的作用,且较雷帕霉素洗脱支架更明显,其凋亡诱导作用和增加局部细胞p53表达有关。     

雷帕霉素涂层支架对支架内早期再狭窄的预防

目的 :评价乳酸和乙醇酸聚合物携带雷帕霉素并以支架为载体抑制血管新生内膜的作用和预防支架内再狭窄的有效性。方法 :采用随机、双盲试验 ,在 2 0头微型猪的冠状动脉前降支或左旋支分别置入支架 1枚 ,其中金属裸支架 8枚 ;雷帕霉素涂层支架 12枚。涂层材料选用乳酸和乙醇酸聚合物 ,根据两种材料比例将涂层支架分为雷帕霉素缓慢释放涂层支架 (药物剂量 6 5～ 90 μg/支架 ,5枚 )和雷帕霉素快速释放涂层支架 (药物剂量 6 8～ 96 μg/支架 ,7枚 )。2 8d后进行冠状动脉造影 ,术后处死动物 ,取出支架血管 ,进行组织学分析。 结果 :对照组的再狭窄率为 2 5 .0 % (2 /8) ;两组雷帕霉素涂层支架均为 0 %。平均狭窄程度 :对照组为 (31± 2 2 ) % ,雷帕霉素缓慢释放涂层支架组减少了 2 8% (P <0 .0 5 ) ;雷帕霉素快速释放涂层支架组减少了 2 3% (P <0 .0 5 ) ;新生内膜面积 :对照组 (2 .18± 1.0 3)mm2 ;雷帕霉素缓慢释放涂层支架组减少了 1.0 9mm2 (P <0 .0 5 ) ;雷帕霉素快速释放涂层支架组减少了 1.2 4mm2 (P <0 .0 5 )。结论 :乳酸和乙醇酸聚合物携带雷帕霉素涂层支架可以降低支架内的狭窄程度 ,有效地减少血管内新生内膜面积 ,预防支架内的狭窄     

NADPH氧化酶与Rho/Rho-激酶激活在外膜介导的血管重塑中的作用

目的　探讨NADPH氧化酶与Rho/Rho 激酶在外膜介导的血管重塑中的作用。方法在体去除兔颈动脉外膜 ,于术后即刻、1周及 2周取出颈动脉。用图像分析系统测定内膜及中层面积 ;应用多道生理仪测血管反应性 ;原位杂交及RT PCR检测mRNA表达。结果　 (1)血管外膜去除后内膜增生 ,2周时内膜增生较 1周时更为明显。对照侧无内膜增生。 (2 )与对照侧比较 ,去外膜侧血管在术后即刻、1周对去甲肾上腺素的收缩反应减弱 (P <0 0 5)。 (3 )与对照侧比较 ,去外膜侧NADPH氧化酶P2 2PhoxmRNA表达在 1周时增高 (P <0 0 5)。 (4)与对照侧比较 ,去外膜侧Rho 激酶mRNA表达在 2周时增高 (P <0 0 5)。结论　外膜去除后血管内膜增生 ,血管舒缩功能改变 ,其发生机制与NADPH氧化酶及Rho 激酶激活有关     

聚丙交酯-乙交酯涂层-钴基合金雷帕霉素洗脱支架对猪冠状动脉新生内膜增生的影响

目的:评价钴基合金支架平台、聚丙交酯-乙交酯(PLGA)聚合物作为携带雷帕霉素涂层的新型支架的抗内膜增殖的有效性及安全性。方法:随机在7头微型猪的3支冠状动脉置入钴基合金PLGA涂层支架(Cob-POS组)、钴基合金PLGA涂层雷帕霉素洗脱支架(Cob-SES组)、不锈钢雷帕霉素药物洗脱支架(gen1-SES组),记录支架释放前后的冠状动脉造影图像。3个月后,冠状动脉造影复查后处死动物,分离支架段血管行组织病理学分析。结果:6只动物存活,1只动物于支架释放过程中死亡,死因可能为麻醉剂所致的呼吸抑制。3组(n=6)支架段血管组织学评价示,与gen1-SES组比较,Cob-SES组新生内膜面积和最大内膜厚度均明显减少。Cob-POS和Cob-SES组支架内狭窄程度与gen1-SES组比较,均差异有统计学意义(P<0.05)。组织形态学示3组支架段血管损伤积分、炎症积分、再内皮化积分差异无统计学意义。结论:在猪冠状动脉支架模型中,钴基合金平台、PLGA涂层的支架设计显示出良好的生物相容性和安全性;携带雷帕霉素的这种支架显示出比第一代雷帕霉素支架更佳的抑制内膜增生的能力。     

Rho-kinase inhibitor suppressed restenosis in porcine coronary balloon angioplasty

BACKGROUND: Constrictive remodeling is thought to be more important than neointimal formation in coronary restenosis after balloon angioplasty. The inhibition of Rho-kinase prevents neointimal proliferation, but now this inhibition that affects constrictive remodeling remains unknown. To explore this issue further, we investigated whether a specific Rho-kinase inhibitor, Y-27632, could suppress restenosis after coronary balloon angioplasty in a porcine model. METHODS: Balloon angioplasty with local administration of Y-27632 (Y group) or vehicle (C group) was performed at 2 and 3 weeks after overstretch injury in a porcine coronary artery. Quantitative coronary angiography (QCA) and quantitative coronary ultrasound (QCU) were performed to assess the coronary lesion segment. A morphometrical analysis was performed in a histological study. Proliferative cells and p27(Kip1)-positive cells were evaluated in the arterial wall using immunohistochemistry. RESULTS: QCA and QCU demonstrated that the minimal lumen diameter and minimal lumen area were greater, and % stenosis was less in the Y group than in the C group. The QCU analysis also revealed a significant inhibition in the increase of the intimal area and a prevention of constrictive remodeling by Y-27632. In the histological study, the intimal, adventitial and collagen areas were significantly smaller in the Y group than in the C group. The Y group also exhibited significantly less proliferative activity and a significantly higher percentage of cells expressing p27(Kip1) in the arterial wall. CONCLUSION: Local delivery of Y-27632 suppressed constrictive remodeling as well as neointimal formation after coronary balloon angioplasty in pigs.     

Long-term inhibition of Rho-kinase induces a regression of arteriosclerotic coronary lesions in a porcine model in vivo

OBJECTIVE: We recently demonstrated that Rho-kinase/ROK/ROCK is functionally upregulated at the arteriosclerotic coronary lesions and plays a key role for coronary vasospastic responses in our porcine model with interleukin (IL)-1beta. In the present study, we tested our hypothesis that Rho-kinase is involved in the pathogenesis of coronary arteriosclerosis per se in our porcine model. METHODS: Segments of the left porcine coronary artery were chronically treated from the adventitia with IL-1beta. Two weeks after the procedure, coronary stenotic lesions with constrictive remodeling and vasospastic response to serotonin were noted at the IL-1beta-treated site, as previously reported. Then, animals were randomly divided into two groups; one group was treated with fasudil for 8 weeks followed by 1 or 4 weeks of washout period and another group served as a control. After oral absorption, fasudil is metabolized to hydroxyfasudil that is a specific inhibitor of Rho-kinase. RESULTS: In the fasudil group, coronary stenosis and vasospastic response were progressively reduced in vivo, while the coronary hyperreactivity was abolished both in vivo and in vitro. Furthermore, Western blot analysis showed that in the fasudil group, the Rho-kinase activity (as evaluated by the extent of phosphorylation of myosin binding subunit of myosin phosphatase, one of the major substrates of Rho-kinase) was significantly reduced, while histological examination demonstrated a marked regression of the coronary constrictive remodeling. CONCLUSIONS: These results indicate that Rho-kinase is substantially involved in constrictive remodeling and vasospastic activity of the arteriosclerotic coronary artery, both of which could be reversed by long-term inhibition of the molecule in vivo. Thus, Rho-kinase may be regarded as a novel therapeutic target for arteriosclerotic vascular disease.     

Effect of glucocorticoids on the biologic activities of myeloma cells: inhibition of interleukin-1 beta osteoclast activating factor-induced bone resorption

Regulatory effects of glucocorticoids (dexamethasone) on myeloma cells as well as bone resorption in multiple myeloma were investigated. Glucocorticoids significantly inhibited proliferation of myeloma cells, and decreased the messenger RNA (mRNA) expressions of interleukin-6 (IL-6) and secretory type immunoglobulin G (IgG). The inhibitory effects of glucocorticoids on myeloma cell proliferation could be due to the decreased expression of IL-6 mRNA, decreased IL-6 production, and thus suppression of autocrine growth by IL-6, which is an autocrine growth factor for myeloma cells as reported previously (Nature 332:83, 1988). Glucocorticoids also inhibited M-protein secretion by decreasing the levels of secretory type Ig mRNA. On the other hand, because IL-1 beta rather than lymphotoxin is considered to be a major osteoclast activating factor (OAF) produced by myeloma cells, and glucocorticoids decreased the expression of IL-1 beta mRNA and markedly suppressed the bone resorbing activity induced by IL-1 beta OAF in 45Ca-release bone resorption assay, it is suggestive that glucocorticoids could inhibit bone resorption induced by IL-1 beta OAF in multiple myeloma. Therefore, from these data it is concluded that glucocorticoids could be more effective chemotherapeutic agents in multiple myeloma than we expected, especially with regards to the inhibitory effects on proliferation and M-protein secretion from myeloma cells, as well as bone resorption by myeloma cells.     

雷帕霉素对高糖诱导肾小球系膜细胞增殖的影响

目的 探讨雷帕霉素对高糖诱导肾小球系膜细胞（GMC）增殖的影响及其可能涉及的途径。方法高糖培养GMC,以不同剂量雷帕霉素干预,MTT及流式细胞仪等方法观察雷帕霉素对细胞增殖和周期的影响,RT-PCR、Western印迹观察细胞Cyclin D1、CyclinE和p27^KIP1的变化。结果高糖刺激下,GMC增殖明显;雷帕霉素能抑制这一作用,且呈剂量依赖性,并下调CyclinD1、CyclinE的基因及蛋白水平,上调p27^KIP1的蛋白表达;与对照组相比,高糖组G0／G1期细胞减少,S期细胞比例增加（P均＜0．05）;雷帕霉素干预后,G0／G1期细胞升高,S期细胞比例减少（P均＜0．05）。结论雷帕霉素可抑制高糖状态下GMC的增殖,且呈剂量依赖性;其可能机制是通过下调CyclinD1、CyclinE及上调p27^KIP1,参与了G1／S期阻滞。     

Inflammatory stimuli upregulate Rho-kinase in human coronary vascular smooth muscle cells

Recent studies have demonstrated that upregulated Rho-kinase plays an important role in the pathogenesis of arteriosclerosis and vasospasm in both animals and humans. However, little is known about the molecular mechanism(s) involved in the Rho-kinase upregulation. Since inflammatory mechanisms have been implicated in the pathogenesis of arteriosclerosis and vasospasm, we examined whether inflammatory stimuli upregulate Rho-kinase in vitro and in vivo. In cultured human coronary vascular smooth muscle cells (hcVSMC), inflammatory stimuli, such as angiotensin II and interleukin-1beta, increased Rho-kinase expression (at both mRNA and protein levels) and function (as evaluated by the extent of the phosphorylation of the ERM (the ezrin/radixin/moesin) family, substrates of Rho-kinase) in a time- and concentration-dependent manner. The expression of Rho-kinase was inhibited by blockades of protein kinase C (PKC) (by either GF109253 or prolonged treatment with phorbol myristate acetate for 24 h) and an adenovirus-mediated gene transfer of dominant-active Ikappa-B, suggesting an involvement of PKC and NF-kappaB in the intracellular signal transduction pathway for the Rho-kinase expression. Furthermore, coronary vascular lesion formation (characterized by medial thickening and perivascular fibrosis) induced by a long-term administration of angiotensin II was markedly suppressed in NF-kappaB(-/-) mice with reduced expression and activity of Rho-kinase in vivo. These results indicate that the expression and function of Rho-kinase are upregulated by inflammatory stimuli (e.g. angiotensin II and IL-1beta) in hcVSMC with an involvement of PKC and NF-kappaB both in vitro and in vivo.     

腺病毒介导组织因子途径抑制物基因转染兔损伤颈动脉后的表达及对内膜增生的抑制作用

目的 检测人组织因子途径抑制物(tissue factor pathway inhibitor,TFPI)基因腺病毒载体系统在体表达及对兔颈动脉球囊损伤后新生内膜增生的抑制作用.方法 共采用中国大耳白兔70只.分为4组,分别为腺病毒介导TFPI基因(Ad-TFPI)转染组、腺病毒介导LacZ基因(Ad-LacZ)转染组、PBS组及假手术组,前3组分别于颈动脉球囊损伤后局部转染Ad-TFPI、Ad-LacZ或PBS,假手术组只分离颈总动脉,不做球囊损伤.Ad-TFPI组和Ad-LacZ组每组25只动物,PBS组及假手术组每组10只.Ad-TFPI组和Ad-LacZ组分别于术后3、7、10、14和28 d各处死3只动物,用RT-PCR、ELISA方法 ,检测TFPI在颈动脉壁中的表达.术后4周时,4组剩余动物(每组10只)行颈动脉造影检测最小管腔直径,之后取血管标本,观察病理形态学变化,测算出血管新生内膜面积、内膜与中膜的比例(I/ M)、管腔狭窄程度.结果 基因转染后3 d,Ad-TFPI 组检测到TFPI mRNA及蛋白表达,10～14 d为表达高峰,28 d有所下降但仍可检测到,Ad-LacZ组未测到人TFPI的表达.基因转染后4周,颈动脉造影结果 示Ad-TFPI组最小管腔直径明显大于PBS组和Ad-LacZ组[(1.17±0.43)mm比(0.43±0.33)mm、(0.39±0.24)mm],管腔狭窄百分率明显低于PBS组和Ad-LacZ组[(32±8.2)%比(79±10.2)%、(81±13.1)%],P均<0.01.形态学检查分析结果 显示Ad-TFPI组新生内膜面积明显小于PBS组和LacZ组[(0.17±0.03)mm2比(0.73±0.03)mm2、(0.78±0.04)mm2],L/ M值明显小于PBS组和LacZ组(1.02±0.25比2.76±0.33、2.92±0.24),管腔狭窄程度明显低于PBS组和LacZ组[(24.5±2.1)%比(78.2±2.8)%、(81.3±3.2)%],P均<0.01.结论 人TFPI腺病毒表达载体可在体内有效表达,并且对血管成形术后再狭窄有抑制作用.     

通心络对血管外膜损伤后氧化应激反应及内膜病变的作用

目的:探讨通心络对血管外膜损伤后内膜病变的作用及机制。方法: 纯种高血脂新西兰大白兔18只,采用胶原酶消化＋钝性机械分离的方法建立兔颈动脉外膜损伤模型后,随机分为对照组、通心络组和阿托伐他汀组,对照组给予盐水灌胃,另两组分别给予通心络和阿托伐他汀进行干预,共12周。采用HE染色法检查外膜损伤血管的形态变化,用免疫组化染色法测定血管组织中α-actin、CD68及Ⅰ、Ⅲ型胶原蛋白的表达;用实时荧光定量PCR(real-time quantitative PCR,RQ-PCR)技术检测外膜损伤后血管组织中NADPH氧化酶亚单位p22phox mRNA的表达。结果: 与对照组比较,通心络和阿托伐他汀均可显著抑制内膜病变的形成,并明显增加病变中α-actin及Ⅰ、Ⅲ型胶原蛋白的表达(P<0.05),抑制p22phox mRNA的表达(P<0.05)。结论: 通心络及阿托伐他汀均可有效地抑制血管外膜损伤后内膜病变的形成,并增加病变的稳定性,抗氧化应激可能是两种药物作用的机制。     

视黄酸影响大鼠肝星状细胞周期素依赖激酶抑制剂的表达

目的 研究视黄酸影响培养大鼠肝星状细胞周期素依赖激酶抑制剂的表达。方法 分离、培养正常大鼠肝星状细胞、传代细胞经1ng/ml转化生长因子β1刺激后再以1nmol/ml视黄酸处理,然后采用MTT法、免疫细胞化学法、原位杂交技术并结合图像定量分析观察细胞增殖、α-平滑肌肌动蛋白、视黄酸受体β2及p16、p21与p27等基因的表达情况。结果 视黄酸明显抑制星状细胞增殖（41．50％,P＜0．05）、降低α－平滑肌肌动蛋白水平（55．09％,P＜0．05）,并诱导视黄酸受体β2基因的表达。同时,视黄酸增高p16蛋白水平（218．75％,P＜0．05）,促进p21蛋白表达;而两组细胞均未能以免疫细胞化学法检出p27。此外,视黄酸对p16、p21与p27 mRNA水平均无影响。结论 视黄酸抑制由转化生长因子β1介导的大鼠肝星状细胞激活与其上调p16、p21基因转录后表达有关。