Thy 1.1干细胞移植能抑制动脉损伤后内膜增生

目的 探讨大鼠Thy-1.1干细胞局部移植对大鼠颈总动脉球囊损伤后内膜增生的影响,评价干细胞移植对再狭窄的作用.方法 雌性SD大鼠60只随机分为2组,每组30只,即干细胞移植组:于颈总动脉球囊损伤后即刻将约5×105Thy-1.1干细胞注入至损伤血管局部;对照组:颈总动脉球囊损伤后局部注入等量生理盐水.另有30只4～6周龄雄性SD大鼠提供骨髓Thy-1.1干细胞.2组各于术后即刻、3 d、7 d、14 d、28 d取损伤血管段,病理组织学方法 观察细胞增殖情况,原位杂交方法 观察移植细胞的定植、分化情况,RT-PCR方法 分析内皮型一氧化氮合酶(eNOS)的表达情况.结果 干细胞移植组内膜面积低于对照组[I/M,干细胞移植组:2.06±0.28比对照组2.42±0.19,P<0.05],干细胞移植组内皮型一氧化氮合酶(eNOS)的表达明显高于对照组[eNOS mRNA,干细胞移植组:0.44±0.06比对照组0.35±0.02,P<0.05],干细胞移植组原位杂交图片中可见内皮细胞有移植的异体细胞.结论 Thy-1.1干细胞局部移植可促进大鼠颈总动脉球囊损伤后再内皮化,抑制内膜增生,预防血管成形术术后再狭窄.     

Thy1.1干细胞移植能抑制动脉损伤后内膜增生

目的探讨大鼠 Thr-1.1干细胞局部移植对大鼠颈总动脉球囊损伤后内膜增生的影响,评价干细胞移植对再狭窄的作用。方法雌性 SD 大鼠60只随机分为2组,每组30只,即干细胞移植组:于颈总动脉球囊损伤后即刻将约5×10~6 Thy-1.1干细胞注入至损伤血管局部;对照组:颈总动脉球囊损伤后局部注入等量生理盐水。另有30只4～6周龄雄性 SD 大鼠提供骨髓 Thy-1.1干细胞。2组各于术后即刻、3 d、7 d、14 d、28 d 取损伤血管段,病理组织学方法观察细胞增殖情况,原位杂交方法观察移植细胞的定植、分化情况,RT-PCR 方法分析内皮型一氧化氮合酶(eNOS)的表达情况。结果干细胞移植组内膜面积低于对照组[I/M,干细胞移植组:2.06±0.28比对照组2.42±0.19,P<0.05],干细胞移植组内皮型一氧化氮合酶(eNOS)的表达明显高于对照组[eNOS mRNA,干细胞移植组:0.44±0.06比对照组0.35±0.02,P<0.05],干细胞移植组原位杂交图片中可见内皮细胞有移植的异体细胞。结论 Thy-1.1干细胞局部移植可促进大鼠颈总动脉球囊损伤后再内皮化,抑制内膜增生,预防血管成形术术后...     

G-CSF不同处理方式对大鼠颈总动脉球囊损伤后内膜增生的影响

目的探讨粒细胞集落刺激因子（G-CSF）不同处理方式对大鼠颈总动脉球囊损伤后内膜增生以及再内皮化的影响,评价对损伤血管的影响。方法将雄性SD大鼠90只随机分为3组,每组30只,即G-CSF预处理组,于颈总动脉球囊损伤前7d开始皮下注射G-CSF,连续7d后,进行颈总动脉球囊导管损伤;G-CSF后处理组,于颈总动脉球囊损伤后即刻开始皮下注射G-CSF,连续7d;对照组:只进行颈总动脉球囊损伤。3组大鼠均于术后即刻,3,7,14,21及28d,取损伤血管段进行病理组织学检查观察细胞增殖的情况,并采用RT-PCR法检测内皮型一氧化氮合酶（eNOS）的表达。结果G-CSF预处理组内膜面积小于G-CSF后处理组及对照组,管腔直径多于对照组,内膜、中膜面积比值在3组中最低。G-CSF预处理组中eNOS的表达明显高于对照组（P<0.05）;G-CSF后处理组中eNOS的表达与对照组比较无明显差异;G-CSF预处理组与G-CSF后处理组比较亦无统计学差异。结论G-CSF可促进大鼠颈总动脉球囊损伤后再内皮化的进程,抑制内膜增生过程,对球囊损伤具有修复作用,可预防血管成形术术后的再狭窄,G-CSF预处理组较G-CSF后处理组的效果更佳。     

粒细胞集落刺激因子对大鼠颈总动脉损伤后新生内膜增生的影响

目的探讨粒细胞集落刺激因子（G-CSF）对动脉粥样硬化（AS）大鼠颈总动脉球囊损伤后再狭窄的影响。方法Wistar大鼠48只,随机分为3组:普通饮食组（普食组,n=6）,高脂饮食组（高脂组,n=42）。高脂组又分为:G-CSF组（n=21）和生理盐水组（NS组,n=21）。高脂饮食后1个月用HE染色及透射电镜证明AS形成情况。G-CSF组和NS组分别于高脂饮食后1个月给予G-CSF或生理盐水干预6d,然后行左颈总动脉球囊损伤术,术后14、21、90d分别用扫描电镜、HE染色及图像分析观察血管内皮形成及内膜的增生情况。结果高脂组大鼠高脂饮食后1个月可见AS的早期改变,行球囊损伤后可见大鼠颈总动脉内膜逐渐增厚,但是G-CSF组明显小于NS组,G-CSF组损伤血管内皮较快恢复连续性,G-CSF组内膜与中膜面积比值（Ai/Am）远小于NS组（P<0.01）。结论G-CSF可以加速AS大鼠损伤血管的再内皮化、抑制血管损伤后再狭窄的发生。     

应用G-CSF动员骨髓干细胞治疗急性心肌梗塞的研究

目的 研究用粒细胞集落刺激因子(G—CSF)动员骨髓干细胞对急性心肌梗塞进行治疗的可行性。方法 将21例急性心肌梗塞患者随机分成治疗组13例和对照组8例,治疗组连续3大给予G—CSF 300μg/天。用QRS记分系统(ORSs)和SPEGT心肌断层显像方法比较两组第6天和30天的心电图记分和缺损面积变化。结果 治疗组第 30天的心电图记分较第6天明显下降(3.01±0.32vs6.79±0.36,P<0.01),核素梗死缺损区面积明显减少(39.2±7.7 vs 56.9±9.1,P<0.05),核素放射性计数百分比明显增加[(25.1±5.3)％ vs (18.1±6.5)％,P<0.05],对照组无明显改变(P>0.05)。结论 用G—CSF动员骨髓干细胞可迁移心脏内,并再生成心肌组织,明显改善梗死心肌。     

G-CSF对造血干细胞的动员机制及对健康供者的影响

粒细胞集落刺激因子（G－CSF）通过影响因干细胞粘附分子的表达及其功能，以及影响骨髋微环境，从而促使骨髓造血干细胞向外周血释放，使外周血造血干细胞移植得以实施。G－CSF可引起供者骨痛，疲乏，头痛、恶心，呕吐，发热等不良反应及免疫功能的暂时性改变，绝大多数供者可耐受上述不良反应，但G－CSF对供者的远期影响尚待进一步研究和观察，本文就上述内容作一综述。     

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

目的 利用干细胞动员剂重组人粒细胞集落刺激因子(rhG-CSF)动员骨髓问充质干细胞(MSCs),探讨其对兔颈动脉粥样硬化球囊损伤后再内皮化和内膜增生过程的影响.方法 制作颈动脉粥样硬化狭窄兔模型67只,分为rhG-CSF组(rhG-CSF+球囊损伤,n=35)和对照组(生理盐水+球囊损伤,n=32).以流式细胞仪检测外周血MSCs数量,苏木精咿红法染色观察损伤动脉形态,免疫组织化学检测增殖细胞核抗原(PCNA).结果 (1)rhG-CSF应用前,两组之间的外周血MSCs细胞数量差异无统计学意义.应用rhG-CSF后24 h外周血MSCs数量显著增加(P＜0.01),第7天达到高峰,14 d时仍明显增高;而对照组各个时间点MSCs数量差别均无统计学意义.(2)形态学观察显示,球囊损伤后1周,对照组内皮细胞仍然大片缺失,仪见少数新生的内皮细胞,而rhG-CSF组内皮细胞散在覆盖,新生内皮排列不规则.14 d时,对照组内皮覆盖面较小,基底层片状裸露,而rhG-CSF组内皮呈小片状覆盖.28 d时rhG-CSF组内皮间连接建立,细胞排列呈现方向性.(3)免疫组织化学检测显示,球囊损伤后,动脉中膜近血管腔侧可见少量PCNA阳性细胞,而新生内膜PCNA 阳性细胞最多.rhG-CSF组于7、14、28 d时,PCNA阳性细胞率(细胞增殖指数)均显著低于对照组(均P＜0.01).结论 rhG-CSF能有效地促进骨髓释放MSCs,提高外周血MSCs数量,并促进损伤动脉再内皮化,抑制新生内膜增生,改善血管重塑.     

G-CSF在心血管疾病方面的临床研究进展

粒细胞集落刺激因子(G-CSF)对心脏有直接保护作用,也可以通过刺激骨髓干细胞动员至外周血,间接发挥作用,加速损伤修复,减少心血管不良事件。近年来涌现出大量关于G-CSF在心血管疾病方面的临床研究文章,G-CSF使用的剂量、时机、疗程、联用药物等均会影响治疗效果,本文总结了国内外的研究结果,可为G-CSF在心血管疾病方面进一步的临床应用提供依据。     

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.     

Role of stem cell factor and granulocyte colony-stimulating factor in remodeling during liver regeneration

Functional pluripotent characteristics have been observed in specific subpopulations of hepatic cells that express some of the known cholangiocyte markers. Although evidence indicates that specific cytokines, granulocyte macrophage colony-stimulating factors (GM-CSFs), and stem cell factors (SCFs) may be candidate treatments for liver injury, the role of these cytokines in intrahepatic biliary epithelium remodeling is unknown. Thus, our aim was to characterize the specific cytokines that regulate the remodeling potentials of cholangiocytes after 70% partial hepatectomy (PH). The expression of the cytokines and their downstream signaling molecules was studied in rats after 70% PH by immunoblotting and in small and large murine cholangiocyte cultures (SMCCs and LMCCs) by immunocytochemistry and real-time polymerase chain reaction (PCR). There was a significant, stable increase in SCF and GM-CSF levels until 7 days after PH. Real-time PCR analysis revealed significant increases of key remodeling molecules, such as S100 calcium-binding protein A4 (S100A4) and miR-181b, after SCF plus GM-CSF administration in SMCCs. SMCCs produced significant amounts of soluble and bound SCFs and GM-CSFs in response to transforming growth factor-beta (TGF-β). When SMCCs were incubated with TGF-β plus anti-SCF+GM-CSF antibodies, there was a significant decrease in S100A4 expression. Furthermore, treatment of SMCCs with SCF+GM-CSF significantly increased matrix metalloproteinases (MMP-2 and MMP-9) messenger RNA as well as miR-181b expression, along with a reduction of metalloproteinase inhibitor 3. Levels of MMP-2, MMP-9, and miR-181b were also up-regulated in rat liver and isolated cholangiocytes after PH. CONCLUSION: Our data suggest that altered expression of SCF+GM-CSF after PH can contribute to biliary remodeling (e.g., post-transplantation) by functional deregulation of the activity of key signaling intermediates involved in cell expansion and multipotent differentiation.     

Functional analysis of human hematopoietic repopulating cells mobilized with granulocyte colony-stimulating factor alone versus granulocyte colony-stimulating factor in combination with stem cell factor

Using in vitro progenitor assays, serum-free in vitro cultures, and the nonobese diabetic/severe combined immune-deficient (NOD/SCID) ecotropic murine virus knockout xenotransplantation model to detect human SCID repopulating cells (SRCs) with multilineage reconstituting function, we have characterized and compared purified subpopulations harvested from the peripheral blood (PB) of patients receiving granulocyte colony-stimulating factor (G-CSF) alone or in combination with stem cell factor (SCF). Mobilized G-CSF plus SCF PB showed a 2-fold increase in total mononuclear cell content and a 5-fold increase in CD34-expressing cells depleted for lineage-marker expression (CD34(+)Lin(-)) as compared with patients treated with G-CSF alone. Functionally, G-CSF plus SCF-mobilized CD34(+)CD38(-)Lin(-) cells contained a 2-fold enhancement in progenitor frequency as compared with G-CSF-mobilized subsets. Despite enhanced cellularity and progenitor capacity, G-CSF plus SCF mobilization did not increase the frequency of SRCs as determined by limiting dilution analysis by means of unfractionated PB cells. Purification of SRCs from these sources demonstrated that as few as 1000 CD34(+)CD38(-)Lin(-) cells from G-CSF-mobilized PB contained SRC capacity while G-CSF plus SCF-mobilized CD34(+)CD38(-)Lin(-) cells failed to repopulate at doses up to 500 000 cells. In addition, primitive CD34(-)CD38(-)AC133(+)Lin(-) cells derived from G-CSF plus SCF-mobilized PB were capable of differentiation into CD34-expressing cells, while the identical subfractions from G-CSF PB were unable to produce CD34(+) cells in serum-free cultures. Our study defines qualitative and quantitative distinctions among subsets of primitive cells mobilized by means of G-CSF plus SCF versus G-CSF alone, and therefore has implications for the utility of purified repopulating cells from these sources.     

Granulocyte colony-stimulating factor after autologous CD34+ immunoselected peripheral blood stem cell transplantation

Granulocyte colony-stimulating factor (G-CSF) can be administered after a peripheral blood stem cell transplantation with the aim of accelerating neutrophil recovery. In a randomized, single-blind study we studied a new administration schedule of G-CSF in this context.     

A prospective randomized trial of granulocyte colony-stimulating factor therapy after autologous blood stem cell transplantation in adults.

In order to assess the potential clinical benefit of filgrastim (G-CSF) after peripheral blood stem cell (PBSC) autotransplantation a randomized study was begun in our center in July 1997: 62 patients were involved (30 received filgrastim after PBSC infusion and 32, the control group, received no cytokines). All were adults (median 40 years, range 18-65). Patients with one of three different pathologies were recruited: 28 had advanced breast carcinoma, 23 had lymphomas (12 Hodgkin's disease and 11 non-Hodgkin's lymphoma) and 11 had de novo AML. All of them were transplanted using myeloablative chemotherapy conditioning regimens. G-CSF was administered subcutaneously from day +5 in the treated group at a dose of 5 microg/kg body weight/day. The numbers of CD34+ and mononuclear (MNC) cells infused were similar in each group. Only minor differences regarding the use of G-CSF could be inferred from the analysis of the data. Faster granulocyte engraftment was evident in the treated group (mean of 10 vs 12 days to achieve >0.5 x 109/l granulocytes, P = 0.0008), without differences in incidence and severity of infections, days of fever or duration of antibiotic treatment between groups. There was slightly slower platelet engraftment (mean of 15 days in the group with G-CSF vs 12 days in the other group to achieve >20 x 109/l platelets, P = NS) in this series, but there were no differences in incidence and severity of haemorrhage or platelet transfusion support. Considering the economical costs, the median expenditure per inpatient stay was Eur5961 (range Eur4386-Eur17186) in the G-CSF group compared with Eur5751 (range Eur3676-Eur15640) in the control group (P = 0.47). From our data it could be concluded that for adult patients transplanted with PBSC there is no clear beneficial impact of post-infusion G-CSF administration.     

The endogenous granulocyte colony-stimulating factor response following autologous peripheral blood stem cell transplantation is impaired in patients with myeloma

Granulocyte colony-stimulating factor (G-CSF) levels were studied in 23 patients (10 myeloma, 13 relapsed Hodgkin's disease, non-Hodgkin's lymphoma or germ cell tumours), post autologous peripheral blood stem cell transplantation (PBSCT). The two groups had similar previous chemotherapy and numbers of CD34+ cells transplanted. All patients received G-CSF by injection starting 8 d post transplantation. Twenty out of 23 patients showed raised endogenous levels of G-CSF before cytokine administration. Myeloma patients showed significantly lower levels of endogenous G-CSF than the other patients (0.767 versus 3.262 ng/ml, P < 0.05). Further rises in G-CSF levels were seen following the administration of exogenous G-CSF which then fell, despite ongoing administration of G-CSF, as neutrophil recovery occurred.     

Influence of post-transplant recombinant human granulocyte colony-stimulating factor administration on peritransplant morbidity in patients undergoing autologous stem cell transplantation

This study evaluated of the effect of post-transplant recombinant human granulocyte colony-stimulating factor (rhG-CSF) administration on the parameters of peritransplant morbidity. Three sequential and consecutive cohorts of 20 patients each received either post-transplant rhG-CSF at a dose of 5 micro g/kg/d i.v. in the morning, starting on d 0, d 5, or no rhG-CSF. Patients who received rhG-CSF starting on d 0 and 5 recovered granulocytes more rapidly than those not receiving rhG-CSF (P < 0.001 for ANC >or= 0.5 and 1 x 10(9)/l). RhG-CSF administration was not significantly associated with more rapid platelet engraftment. RhG-CSF administration starting on d 0 and 5 was significantly associated with a decreased duration of fever (P = 0.002 and 0.001 respectively), antibiotic administration (P < 0.001 and 0.006 respectively) and shorter hospitalization (P < 0.001 and 0.001 respectively) compared with the reference group. There was no difference between the d 0 and d 5 arms regarding the parameters of peritransplant morbidity. In conclusion, rhG-CSF administration was associated with a faster granulocyte recovery, shorter hospitalization, and shorter period of fever and non-prophylactic antibiotic administration. This study also showed that starting rhG-CSF administration on d 5 may be as effective as d 0 on the clinical outcome and may be an economical approach in routine clinical practice in this cost-conscious era.     

Effect of granulocyte colony-stimulating factor on bone metabolism during peripheral blood stem cell mobilization

Granulocyte colony-stimulating factor (G-CSF) has been shown to affect the biochemical markers of bone metabolism, including serum bone alkaline phosphatase (BALP), serum osteocalcin, and urine deoxypyridinoline. To determine the association between bone resorption and formation and the G-CSF-induced mobilization of peripheral blood stem cells (PBSC), we examined these markers during mobilization in 19 healthy donors. The average (+/- SEM) serum BALP level before treatment was 81.6 +/- 17.0 IU/dL, and the level increased significantly to 117.7 +/- 15.8 IU/dL on day 5 of G-CSF administration (P < .0001). The urine deoxypyridinoline level before treatment was 12.3 +/- 2.4 nmol/mmol creatinine, and this level also increased significantly to 19.4 +/- 3.0 nmol/mmol creatinine on day 5 of G-CSF administration (P < .0001). In contrast, the average level of serum osteocalcin significantly decreased from 8.07 +/- 2.88 ng/mL to 1.53 +/- 0.18 ng/mL on day 5 (P = .0353). During G-CSF administration, we also studied the serum levels of various cytokines (IL-1beta, osteoclastogenesis inhibitory factor [OCIF], IL-6, tumor necrosis factor alpha, transforming growth factor beta, interferon-gamma, macrophage colony-stimulating factor) related to bone metabolism. Only the kinetics of OCIF were significantly affected. The serum level of OCIF increased immediately after the start of G-CSF administration and remained high during G-CSF administration. These results demonstrate that high-dose G-CSF affects bone metabolism and that OCIF may play a role in bone metabolism. Consistent with the notion that G-CSF affects bone metabolism, a significant correlation was observed between CD34+ cell yield and the increase in urine deoxypyridinoline but not for the changes in serum BALP and osteocalcin levels. This result suggests that bone resorption is either directly or indirectly related to the mobilization of PBSC by G-CSF.