Hirulog-1 reduces expression of platelet-derived growth factor in neointima of rat carotid artery induced by balloon catheter injury

Vascular restenosis is one of the major concerns for the treatment of atherosclerotic cardiovascular diseases using therapeutic vascular procedures. Hirulog-1, a synthetic thrombin inhibitor, effectively reduced ischemic events in coronary heart disease patients and caused less hemorrhagic complications compared to heparin. Thrombin stimulated the expression of platelet-derived growth factor (PDGF) in vascular cells. PDGF receptor blockers reduced angioplasty-induced restenosis in the swine model. The present study examined the effects of hirulog-1 on vascular stenosis, platelet deposition and the expression of PDGF in rat carotid arteries injured by balloon catheter. Multiple intravenous infusions of hirulog-1 (1 mg/kg/h for 4 h for 6 times), but not bolus injection or 1-2 times of infusion, reduced neointima/media ratio by 50% in balloon-injured carotid arteries compared to injured animals receiving saline alone. Activated partial thromboplastin time in hirulog-1-treated rats was significantly prolonged compared to saline controls but shorter than that in animals receiving heparin (50 U/kg/h). One of heparin-treated rat, but none of hirulog-1-treated, died from bleeding complication. Hirulog-1 injection transiently reduced platelet deposition on denuded intima visualized by scanning electron microscopy. Abundance of PDGF in neointima of injured carotid arteries detected by immunohistochemistry was significantly decreased following infusions of hirulog-1. The results suggest that balloon catheter injury induced neointima formation and the overexpression of PDGF in the neointima of rat carotid artery may be effectively suppressed by infusions with hirulog-1, a thrombin-specific inhibitor.     

Roscovitine抑制大鼠颈总动脉球囊损伤后内膜增生的研究

目的:观察roscovitine对球囊损伤后大鼠颈总动脉血管平滑肌细胞及内膜增生的抑制作用,以期提供新型的支架涂层药物。方法:建立大鼠颈总动脉球囊损伤模型模拟经皮冠状动脉腔内介入术(PCI)术后再狭窄,干预组损伤局部给予roscovitine(200μmol/L)孵育10分钟。14天后取材,免疫荧光染色观察roscovitine对局部血管平滑肌细胞增殖的作用;HE染色观察roscovitine对内膜增生的作用。结果:本研究建立了大鼠颈总动脉球囊损伤模型,球囊损伤后14天,局部血管平滑肌细胞增殖活跃、内膜增生明显。Roscovitine干预后,血管平滑肌细胞增殖率明显降低,内膜增生被显著抑制,管腔狭窄率和内膜中膜面积比值显著降低。结论:Roscovitine显著抑制大鼠颈总动脉球囊损伤后局部血管平滑肌细胞增殖,进而有效抑制内膜增生,降低再狭窄发生率。     

High potassium intake inhibits neointima formation in the rat carotid artery balloon injury model

Recently, we reported that elevated extracellular potassium concentration in vitro inhibited proliferation and migration of vascular smooth muscle cells, formation of free radical compounds by macrophages, and reduced platelet sensitivity to agonists. In the present study we analyzed the effects of long-term, in vivo elevation of extracellular potassium concentration resulting from changes in dietary potassium intake on the vascular response to injury. The rat carotid artery balloon injury model was employed in 70 adult Sprague Dawley rats assigned to three groups. Beginning 14 days before surgical placement of the carotid lesion and continuing until death, the animals were fed diets containing either low (0.1% potassium, n = 25), normal (1.5% potassium, n = 19), or high potassium (4.0% potassium, n = 26). Fourteen days postsurgery the animals were killed and the arteries were analyzed to determine quantitatively the ratio of neointimal to medial area. Dietary potassium had a significant effect on arterial plasma potassium concentration (one-way analysis of variance, P < .01). Group mean and standard errors were 4.26+/-0.12 mmol/L for the low-potassium group, 5.22+/-0.19 mmol/L for normal, and 5.80+/-0.23 mmol/L for the high-intake group. Increases in dietary potassium attenuated neointima formation significantly (P < .05, one-way analysis of variance), with the mean ratio of neointimal area to medial area being 0.447+/-0.106 for the low-intake animals, 0.384+/-.116 for normal, and 0.240+/-.046 for the high-intake group. These results are consistent with a hypothesis that a high level of potassium intake is effective in inhibiting neointima formation in vivo.     

Ursolic acid inhibits neointima formation in the rat carotid artery injury model

Triterpenoids are natural compounds that are found in a large variety of plants and vegetarian foods, and are used for medicinal purposes in many Asian countries. Pentacyclic triterpenes, such as ursolic acid, have been reported to exhibit anticancer and anti-inflammatory properties. The present study was designed to assess the effects of ursolic acid in the migration and proliferation of vascular smooth muscle cells (VSMC), and in a vascular injury model. The exposure of VSMC to ursolic acid results in a chemotaxis inhibition, in a reduction of the expression of proliferating cell nuclear antigen (PCNA) and in a disorganization of beta-tubulin and vimentin cytoskeletal proteins. Administration of ursolic acid in the rat carotid balloon catheter injury model shows a significant inhibition of neointimal hyperplasia. Thus, we have demonstrated that daily doses of 6 mg/kg body weight for 10 days reduce both the ratio of intimal to medial areas and the degree of stenosis by 80%, and suppress the expression of PCNA in both neointima and media. These results suggest that pentacyclic triterpenes may be of potential therapeutic value in vascular injury, and a possible treatment strategy for the prevention of the progression of atherosclerosis and restenosis after angioplasty.     

Effects of probucol on rat carotid artery responses to balloon catheter injury

Balloon catheter injury to the rat common carotid artery has been widely used for testing potential therapies for post-angioplasty restenosis. However, the model has become somewhat discredited because a number of drugs that inhibit intimal thickening, measured 14 days after balloon catheter injury, have been found to be ineffective in clinical trials. Probucol has recently been shown to reduce the incidence of post-angioplasty restenosis in a number of small clinical trials, making it possible to reassess the validity of the rat balloon injury model. The effects of probucol on the underlying causes of intimal thickening in balloon-injured rat carotid arteries were quantified. Probucol inhibited medial smooth muscle cell proliferation by 23% on day 4 after injury (P=0.009), and by 65% on day 10 after injury (P=0.026). Smooth muscle cell migration was reduced by 64% (P=0.008) in probucol-treated animals. In marked contrast, intimal smooth muscle cell proliferation was significantly increased by 41% (P=0.024) by probucol. There was no significant effect on intimal thickening or smooth muscle cell death. These data suggest that drugs that inhibit both medial smooth muscle cell proliferation and migration in the rat balloon injury model may prove useful in the treatment of post-angioplasty restenosis.     

Arsenic trioxide eluting stent reduces neointima formation in a rabbit iliac artery injury model

OBJECTIVE: In-stent restenosis is caused by the neointimal hyperplasia, which involves abnormal growth of vascular smooth muscle cells (VSMC). Arsenic trioxide (As2O3) is known to be a potent inhibitor of cell proliferation. We therefore studied the role of an As2O3 eluting stent in the prevention of restenosis in a rabbit iliac artery model. METHODS AND RESULTS: Bare stents, or stents coated with poly-L-lactic acid (PLLA) and either 40 microg of As2O3, 180 microg of paclitaxel or vehicle were implanted into the left proximal iliac arteries of New Zealand rabbits. The delivery of drugs from stents in vitro and in vivo was evaluated by atomic fluorescence spectrophotometry and high-performance liquid chromatography, respectively. Histomorphometric measurements at 7 or 28 days showed that, comparing to rabbits receiving the PLLA stent, in animals treated with As2O3 eluting or paclitaxel eluting stent neointima thickness was reduced by 50% and 46%, the absolute neointimal area was reduced by 53% and 44%, while the absolute luminal area was increased by 46% and 43%, respectively. There were no significant differences in injury or inflammation scores among PLLA, As2O3 eluting and paclitaxel eluting stents. As2O3 eluting stent induced more TUNEL-positiv VSMC than the other stents. As2O3 levels measured in the arterial tissue were much higher than those in serum, which were nearly undetectable at 7 days after stent implantation. In in vitro studies, cultured rabbit arterial VSMC were stimulated with As2O3 or paclitaxel and analyzed for their cell cycle progression and apoptosis by flow cytometry and electron microscopy. As2O3 treatment resulted in a reduction of VSMC number in G1 phase with a concomitant increase in apoptosis of VSMC, whereas paclitaxel treatment led to blocking of VSMC in the G2/M phase. CONCLUSION: In a rabbit iliac artery model PLLA coated As2O3 eluting stent significantly suppressed in-stent restenosis by reducing proliferation and inducing apoptosis of VSMC.     

罗格列酮对大鼠颈动脉球囊损伤后新生内膜及炎症因子的影响

目的观察罗格列酮对大鼠颈动脉损伤后新生内膜及炎症因子的影响。方法将SD大鼠随机分为3组(即对照组、手术组、治疗组)。手术组及治疗组均用球囊导管扩张法损伤左侧颈总动脉,治疗组给予罗格列酮灌胃治疗。术后4h,1d和7d,用放免法检测大鼠血浆中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平。2周后取损伤血管行苏木精-伊红染色、免疫组化染色及光镜观察,计算内膜面积(NIA)、内弹力板围绕面积(IELA)、管腔狭窄指数(SI)及核转录因子-κB(NF-κB)蛋白表达的光密度值。结果罗格列酮能抑制损伤动脉新生内膜的形成,抑制NF-κB的蛋白表达,降低血浆中TNF-α和IL-6水平。结论罗格列酮可以通过抑制损伤血管的炎症反应,减轻损伤血管的再狭窄。     

S100B在大鼠颈总动脉损伤后新生内膜中的表达及意义

目的 探讨S100B在大鼠颈总动脉损伤后新生内膜中表达特征及其临床意义。方法 将体质量400~500 g SD大鼠,随机分为假手术组和颈总动脉球囊损伤模型组,于术后1、3、7、14和28 d取损伤的颈总动脉,40 g/L福尔马林固定过夜,切片行HE染色观察新生内膜形成情况,Western blot、qRT-PCR、免疫组化染色检测S100B的表达。检测干预因素对新生内膜影响时,将大鼠随机分为3组,即假手术组、Ad-GFP组、Ad-shS100B组,假手术组不做球囊拉伤,其余两组除球囊拉伤外分别给予200 μl Ad-GFP和Ad-shS100B。结果 颈总动脉损伤后,新生内膜增生随着时间的延长呈现出逐渐增加的趋势,且在14 d时达峰值（P<0.05）,而内膜与中膜面积比值在28 d达到最大。qRT-PCR显示S100B-mRNA在颈总动脉损伤后呈现出第1天升高,第3、7天时有所下降,第14天时上升,而第28天又下降的动态趋势(P<0.05)。Western blot显示颈总动脉损伤后S100B表达逐渐增加,7 d时达到高峰,随后降低（P<0.05）。免疫组化结果显示增加的S100B蛋白主要存在于新生内膜中。给予Ad-shS100B干预后,新生内膜形成及S100B的表达明显受到抑制。结论 颈总动脉损伤后S100B的表达特征与新生内膜的形成有关。     

Dendritic cells in neointima formation after rat carotid balloon injury: coordinated expression withanti-apoptotic Bcl-2 and HSP47 in arterial repair

OBJECTIVES: We sought to evaluate: 1) the contribution of dendritic cells (DCs); and 2) the impact of B-cell lymphoma 2 protein (Bcl-2), a central anti-apoptotic protooncogene, and of heat shock protein 47 (HSP47), indicating subsequent collagen deposition, in neointima formation after angioplasty. BACKGROUND: The origin of neointimal cells and the factors that promote their accumulation are still unclear. Previous studies reported intimal presence of DCs and suggested cells of primarily extravascular origin to contribute to arterial repair. METHODS: Sprague-Dawley rats underwent carotid balloon angioplasty. At different times after angioplasty, tissue sections were analyzed by immunohistochemistry using OX-62 and S100 as DC markers and antibodies against Bcl-2 and HSP47, supplemented by electron microscopic analysis of cell type and apoptosis. RESULTS: Four days after injury, DCs adhered along the internal elastic lamina and demonstrated intense Bcl-2 and HSP47 expression, consistent with low apoptosis. With ongoing neointima enlargement, luminal DCs remained prevalent and were colocalized with Bcl-2 and HSP47, while signaling decreased to basal regions. Media showed no DCs and only low Bcl-2 and HSP47 immunoreactivity. Adventitia transiently revealed a structural separation between day 4 and 7. Whereas the inner layer demonstrated sparse cellularity, apoptosis and no DC, Bcl-2, and HSP47 labeling, the outer layer was characterized by high myofibroblast density with strong Bcl-2 and HSP47 expression but absence of DCs. CONCLUSIONS: We identify DCs as novel components in early neointima formation, promoted by coordinated anti-apoptotic Bcl-2 and HSP47 expression. Despite intense adventitial remodeling, there is no evidence of adventitial cell transmigration.     

慢病毒载体在大鼠颈动脉球囊损伤模型中的应用及观察

目的:研究携带绿色荧光蛋白(Green fluorescent protein,GFP)的慢病毒转染球囊损伤大鼠颈动脉的效率和可行性,为利用慢病毒载体介导的基因治疗预防血管再狭窄奠定基础。方法:将携带GFP的慢病毒载体(pGC-LV-GFP载体)和慢病毒包装质粒供转染293T细胞,完成慢病毒颗粒包装及滴度测定。包装产生的慢病毒Lenti-GFP转染至球囊损伤的大鼠颈动脉。术后28d,损伤及病毒转染血管段标本分别行荧光显微镜、光镜检查,评估慢病毒的转染效率及内膜增生情况,并与假手术组(Sham组)、PBS对照组进行比较。结果:经包装产生的Lenti-GFP滴度为2×109TU/mL;术后28d,Sham组与PBS组血管中膜弹力层仅见微弱的自发性绿色荧光,而Lenti-GFP组动脉壁全层可见较强的绿色荧光分布;PBS组与Lenti-GFP转染组大鼠损伤的颈动脉均可见明显内膜增生,内膜和中膜面积之比差异无统计学意义。结论:Lenti-GFP成功转染至球囊损伤的大鼠颈动脉,并能维持目的基因稳定长期的表达,是血管再狭窄基因治疗的理想载体。     

Insulin stimulates arterial neointima formation in normal rats after balloon injury

AIM: Insulin resistance in patients is associated with increased atherosclerosis and arterial restenosis. It is thought that the concomitant hyperinsulinaemia exacerbates vascular disease because resistance to insulin-induced glucose disposal is associated with resistance to certain effects of insulin which inhibit, but with no resistance to other effects which promote, neointimal hyperplasia. We sought to determine the net effect of hyperinsulinaemia on neointimal hyperplasia in normal animals. METHODS: Rats were infused with or without insulin for 16 days and the carotid artery injured by balloon catheter on day 2. RESULTS: Steady-state serum insulin concentrations were 0.56 +/- 0.04 and 1.25 +/- 0.05 nm for control and hyperinsulinaemic rats respectively (p < 0.01). Systolic blood pressures, weights and serum glucose levels were not affected by hyperinsulinaemia. Fourteen days after injury, the neointima-to-media area ratio was 0.72 +/- 0.07 and 1.39 +/- 0.15 for control and hyperinsulinaemic rats respectively (p < 0.05). Media area was unaffected by hyperinsulinaemia. CONCLUSIONS: The effects of hyperinsulinaemia which promote neointimal hyperplasia after balloon injury of rat carotid artery predominate over the effects which inhibit it even in normal animals.     

AT2R基因转染对血管平滑肌细胞增殖核抗原表达的影响

目的:探讨血管紧张素Ⅱ2型受体(AT2R)基因体外及体内转染对血管平滑肌细胞(VSMC)增殖核抗原(PCNA)表达的影响。方法:细胞爬片及大鼠颈动脉球囊损伤后,局部转染带有绿色荧光蛋白报告基因的AT2R重组腺病毒载体(pAd-AT2R)或病毒载体(pAd-GFP),于细胞转染后48h及术后14d取材,应用免疫组织化学染色、免疫荧光双标染色和激光共聚焦技术检测了VSMC及血管中AT2R与PCNA的表达关系。结果:pAd-AT2R转染VSMC后,在激光共聚焦显微镜下胞质及胞核有绿色荧光蛋白表达;同时在胞质有红色荧光AT2R表达,胞核无蓝色荧光,PCNA表达阴性,无红色荧光AT2R表达的VSMC胞核有蓝色荧光,PCNA表达阳性。pAd-AT2R在体转染大鼠颈动脉后,新生内膜及中膜中PCNA阳性表达率显著低于未转染组和pAd-GFP组[(27.29±5.81)%∶(72.25±4.47)%,(68.43±9.12)%,P<0.01];在激光共聚焦显微镜下,绿色荧光及红色荧光AT2R主要分布于新生内膜、中膜、外膜,内膜散在蓝色荧光PCNA表达,在红、绿色荧光处无蓝色荧光,PCNA表达阴性,无红、绿色荧光处,有蓝色荧光,PCNA表达阳性。结论:AT2R基因体外及体内转染可抑制VSMCPCNA表达,ATR基因转染能抑制新生内膜增生。     

Bradykinin B(1) receptor mediates inhibition of neointima formation in rat artery after balloon angioplasty

We evaluated the effects of the kallikrein-kinin system on the proliferation and migration of primary cultured vascular smooth muscle cells (VSMCs) in vitro and neointima formation in balloon-injured rat carotid arteries in vivo. In cultured rat VSMCs, tissue kallikrein inhibited cell proliferation, and this inhibitory effect was blocked by Sar-Tyr-Aca(epsilon)-Lys [D-betaNal(7), Ile(8)]-des-Arg(9)-bradykinin, a bradykinin B(1) receptor antagonist, and by icatibant, a bradykinin B(2) receptor antagonist. Platelet-derived growth factor significantly increased the expression of the B(1) receptor but not the B(2) receptor in VSMCs. Platelet-derived growth factor-induced cell migration was significantly attenuated by des-Arg(9)-bradykinin and to a lesser degree by bradykinin. Endogenous B(1) receptor mRNA increased in rat carotid arteries after balloon angioplasty. After local delivery of adenovirus carrying the human tissue kallikrein gene into the rat carotid artery, we observed a 54% reduction in the intima/media ratio at the injured site compared with the control ratio (n=7, P:<0.01). Administration of the B(1) receptor antagonist via minipumps blocked the protective effect of kallikrein and partially reversed the intima/media ratio toward the control ratio. Kallikrein gene delivery results in the regeneration of endothelium compared with the control groups, and the B(1) receptor antagonist abolished this effect. Nitrite/nitrate, cGMP, and cAMP levels in balloon-injured arteries significantly increased after kallikrein gene delivery, whereas the B(1) receptor antagonist abolished these increases (n=4 or 5, P:<0.05). These results indicate that the B(1) receptor contributes to the reduction of neointima formation via the promotion of reendothelialization and inhibition of VSMC proliferation and migration through NO-cGMP and cAMP signaling pathways. This study provides significant implications in treating restenosis after revascularization.     

重组人粒细胞集落刺激因子对兔颈动脉粥样硬化球囊损伤后再内皮化和内膜增生的影响

Objeelive To investigate the effect of rhG-CSF on mobilizing bone marrow-MSCs, reendothelialization and intima hyperplasia in carotid artery of rabbits post balloon catheter injury, nethods Rabbits were treated with rhG-CSF (25 μkg, twice daily, i. p, n =35) or saline (n =32) for 5 days, then, carotid arteries of rabbits were injured by balloon catheter. The number of peripheral MSCs was detected with FACS. The morphology of injuried artery was examined with hematoxylin and eosin stain, PCNA was determined with immunohistochemistry. Results (1) Number of peripheral MSCs was similar at baseline and significantly increased at 24 hours and peaked at 7 days and remained increased till 14 days post rhG-CSF. (2) Significant endothelial cell deletion was evidenced in the control group, while scatter endothelial cells was observed in the rhG-CSF group at 1 week post injury. Two weeks after injury, new endothelial area was significantly higher in rhG-CSF group compared to control group. At 4 weeks post injury, endothelial connection was evidenced and regularly displayed in rhG-CSF treated group. (3) PCNA-positive cells in the tunica intima were significantly lower in rhG-CSF treated rabbits at 7, 14 and 28 days compared that in control rabbits (all P ＜ 0.01). Conclusion rhG-CSF could mobilize the bone marrow-MSCs and promote re-endothelialization and attenuate intima hyperplasia post balloon catheter injury in carotid arteries of rabbits.     

重组人粒细胞集落刺激因子对兔颈动脉粥样硬化球囊损伤后再内皮化和内膜增生的影响

Objeelive To investigate the effect of rhG-CSF on mobilizing bone marrow-MSCs, reendothelialization and intima hyperplasia in carotid artery of rabbits post balloon catheter injury, nethods Rabbits were treated with rhG-CSF (25 μkg, twice daily, i. p, n =35) or saline (n =32) for 5 days, then, carotid arteries of rabbits were injured by balloon catheter. The number of peripheral MSCs was detected with FACS. The morphology of injuried artery was examined with hematoxylin and eosin stain, PCNA was determined with immunohistochemistry. Results (1) Number of peripheral MSCs was similar at baseline and significantly increased at 24 hours and peaked at 7 days and remained increased till 14 days post rhG-CSF. (2) Significant endothelial cell deletion was evidenced in the control group, while scatter endothelial cells was observed in the rhG-CSF group at 1 week post injury. Two weeks after injury, new endothelial area was significantly higher in rhG-CSF group compared to control group. At 4 weeks post injury, endothelial connection was evidenced and regularly displayed in rhG-CSF treated group. (3) PCNA-positive cells in the tunica intima were significantly lower in rhG-CSF treated rabbits at 7, 14 and 28 days compared that in control rabbits (all P ＜ 0.01). Conclusion rhG-CSF could mobilize the bone marrow-MSCs and promote re-endothelialization and attenuate intima hyperplasia post balloon catheter injury in carotid arteries of rabbits.     

重组人粒细胞集落刺激因子对兔颈动脉粥样硬化球囊损伤后再内皮化和内膜增生的影响

Objeelive To investigate the effect of rhG-CSF on mobilizing bone marrow-MSCs, reendothelialization and intima hyperplasia in carotid artery of rabbits post balloon catheter injury, nethods Rabbits were treated with rhG-CSF (25 μkg, twice daily, i. p, n =35) or saline (n =32) for 5 days, then, carotid arteries of rabbits were injured by balloon catheter. The number of peripheral MSCs was detected with FACS. The morphology of injuried artery was examined with hematoxylin and eosin stain, PCNA was determined with immunohistochemistry. Results (1) Number of peripheral MSCs was similar at baseline and significantly increased at 24 hours and peaked at 7 days and remained increased till 14 days post rhG-CSF. (2) Significant endothelial cell deletion was evidenced in the control group, while scatter endothelial cells was observed in the rhG-CSF group at 1 week post injury. Two weeks after injury, new endothelial area was significantly higher in rhG-CSF group compared to control group. At 4 weeks post injury, endothelial connection was evidenced and regularly displayed in rhG-CSF treated group. (3) PCNA-positive cells in the tunica intima were significantly lower in rhG-CSF treated rabbits at 7, 14 and 28 days compared that in control rabbits (all P ＜ 0.01). Conclusion rhG-CSF could mobilize the bone marrow-MSCs and promote re-endothelialization and attenuate intima hyperplasia post balloon catheter injury in carotid arteries of rabbits.     

重组人粒细胞集落刺激因子对兔颈动脉粥样硬化球囊损伤后再内皮化和内膜增生的影响

Objeelive To investigate the effect of rhG-CSF on mobilizing bone marrow-MSCs, reendothelialization and intima hyperplasia in carotid artery of rabbits post balloon catheter injury, nethods Rabbits were treated with rhG-CSF (25 μkg, twice daily, i. p, n =35) or saline (n =32) for 5 days, then, carotid arteries of rabbits were injured by balloon catheter. The number of peripheral MSCs was detected with FACS. The morphology of injuried artery was examined with hematoxylin and eosin stain, PCNA was determined with immunohistochemistry. Results (1) Number of peripheral MSCs was similar at baseline and significantly increased at 24 hours and peaked at 7 days and remained increased till 14 days post rhG-CSF. (2) Significant endothelial cell deletion was evidenced in the control group, while scatter endothelial cells was observed in the rhG-CSF group at 1 week post injury. Two weeks after injury, new endothelial area was significantly higher in rhG-CSF group compared to control group. At 4 weeks post injury, endothelial connection was evidenced and regularly displayed in rhG-CSF treated group. (3) PCNA-positive cells in the tunica intima were significantly lower in rhG-CSF treated rabbits at 7, 14 and 28 days compared that in control rabbits (all P ＜ 0.01). Conclusion rhG-CSF could mobilize the bone marrow-MSCs and promote re-endothelialization and attenuate intima hyperplasia post balloon catheter injury in carotid arteries of rabbits.     

重组人粒细胞集落刺激因子对兔颈动脉粥样硬化球囊损伤后再内皮化和内膜增生的影响

Objeelive To investigate the effect of rhG-CSF on mobilizing bone marrow-MSCs, reendothelialization and intima hyperplasia in carotid artery of rabbits post balloon catheter injury, nethods Rabbits were treated with rhG-CSF (25 μkg, twice daily, i. p, n =35) or saline (n =32) for 5 days, then, carotid arteries of rabbits were injured by balloon catheter. The number of peripheral MSCs was detected with FACS. The morphology of injuried artery was examined with hematoxylin and eosin stain, PCNA was determined with immunohistochemistry. Results (1) Number of peripheral MSCs was similar at baseline and significantly increased at 24 hours and peaked at 7 days and remained increased till 14 days post rhG-CSF. (2) Significant endothelial cell deletion was evidenced in the control group, while scatter endothelial cells was observed in the rhG-CSF group at 1 week post injury. Two weeks after injury, new endothelial area was significantly higher in rhG-CSF group compared to control group. At 4 weeks post injury, endothelial connection was evidenced and regularly displayed in rhG-CSF treated group. (3) PCNA-positive cells in the tunica intima were significantly lower in rhG-CSF treated rabbits at 7, 14 and 28 days compared that in control rabbits (all P ＜ 0.01). Conclusion rhG-CSF could mobilize the bone marrow-MSCs and promote re-endothelialization and attenuate intima hyperplasia post balloon catheter injury in carotid arteries of rabbits.