粒细胞集落刺激因子对大鼠脑出血后脑水肿及MMP-9的影响

目的探讨粒细胞集落刺激因子(G-CSF)对大鼠脑出血(ICH)后脑水肿的影响及可能机制。方法健康雄性SD大鼠随机分为假手术组、模型组、治疗组,断尾取自体血法制备大鼠ICH模型,治疗组于造模1小时后腹腔注射重组粒细胞集落刺激因子(rhG-CSF)60μg/kg。3组大鼠于术后6、12、24、48、72h和7d处死,干湿重法测定脑组织含水量,免疫组化染色观察MMP-9的动态表达。结果模型组脑组织含水量明显高于假手术组(P<0.05),在造模后48、72h最明显,且与6、12、24h和7d相比差异显著(P<0.05);治疗组脑组织含水量明显低于模型组(P<0.05)。模型组脑组织MMP-9的表达明显高于假手术组(P<0.05);治疗组各时间点MMP-9的表达低于模型组(P<0.05)。结论 G-CSF可减轻ICH后脑水肿程度,其机制可能与G-CSF抑制MMP-9表达密切相关。     

粒细胞集落刺激因子对脑出血大鼠神经行为学的影响

目的:观察粒细胞集落刺激因子(G-CSF)对脑出血大鼠的神经行为学的影响。方法:胶原酶定位注射构建脑出血大鼠模型,然后给予G-CSF腹腔注射连续5d,通过肢体置放实验检测大鼠神经功能恢复情况。结果:术后给予G-CSF的大鼠肢体置放实验评分明显高于未给药组。结论:G-CSF能够促进脑出血大鼠的神经功能的恢复。     

粒细胞集落刺激因子刺激脑出血大鼠星形胶质细胞的增殖

目的:研究粒细胞集落刺激因子(G-CSF)对脑出血大鼠神经保护作用的机制。方法:胶原酶定位注射构建脑出血大鼠模型,然后给予G-CSF腹腔注射连续5d,用免疫组化的方法分别检测实验组与阴性对照组的星形胶质细胞。结果:G-CSF治疗组大鼠出血灶周围星形胶质细胞密度高于对照组。结论:G-CSF可增强脑出血后星形胶质细胞的增殖,这种增殖可能参与粒细胞集落刺激因子对脑出血的保护作用。     

粒细胞集落刺激因子促进大鼠脑出血后的细胞增殖

目的研究粒细胞集落刺激因子（G-CSF）对脑出血神经保护作用的机制。方法胶原酶定位注射构建脑出血大鼠模型,然后给予G-CSF腹腔注射连续5天,造模2周后通过免疫组化检测出血灶周边组织PCNA阳性细胞与GFAP阳性细胞的表达情况。结果给予G-CSF的大鼠PCNA阳性细胞与GFAP阳性细胞计数均明显高于未给药组。结论G-CSF能够促进大鼠脑出血后细胞（尤其是星形胶质细胞）的增殖,该作用可能参与其对脑出血大鼠的神经保护作用。     

粒细胞集落刺激因子在脑出血大鼠中的抗凋亡作用

目的:研究粒细胞集落刺激因子(G-CSF)对脑出血大鼠神经保护作用的机制。方法:胶原酶定位注射构建脑出血大鼠模型,然后给予G-CSF腹腔注射连续5d,用Hoechst33258荧光染色的方法检测G-CSF的抗凋亡作用。结果:术后给予G-CSF的大鼠出血灶所在大脑半球神经细胞凋亡明显低于未给药组。结论:G-CSF在脑出血大鼠脑内具有抗凋亡作用,该作用可能参与其对脑出血大鼠的神经保护作用。     

重组人粒细胞集落刺激因子对大鼠实验性脑出血急性期保护作用的研究

目的:通过观察重组人粒细胞集落刺激因子(rhG-CSF)对脑出血后血肿周围组织胶质纤维酸性蛋白(GFAP)表达及脑组织含水量变化的影响,探讨rhG-CSF在脑出血后继发性损伤中的保护作用。方法:实验动物随机分为假手术组、脑出血组、治疗组。利用鼠脑立体定向仪、采用断尾取自体血法制模,治疗组于制模后1h腹腔注射rhG-CSF60μg/kg。在各时间点将大鼠处死,免疫组化观察各组GFAP在出血后不同时间点的表达;干湿比重法定量测定脑组织含水量。结果:脑出血后脑组织含水量逐渐增加,72h达高峰,各时间点与假手术组相比差异显著;GFAP在脑出血6h后即已开始表达,随后持续增加,72h达高峰。治疗组较相同时间点脑出血组血肿周围组织脑组织含水量降低;GFAP表达有所下降。直线相关分析表明脑出血后1周内GFAP的变化与脑组织含水量变化呈正相关关系。结论:rhG-CSF能够抑制GFAP过度表达、减轻脑出血后脑水肿,对出血后脑组织具有保护作用。     

重组人粒细胞集落刺激因子对实验性脑出血大鼠的神经保护作用

目的:探讨重组人粒细胞集落刺激因子(rhG-CSF)动员骨髓干细胞(BMSC)对大鼠脑出血(ICH)后脑水肿、新生血管的影响。方法:实验动物随机分为假手术组、ICH组、治疗组。利用鼠脑立体定向仪、采用断尾取自体血法制模型,治疗组于制模后1h腹腔注射rhG-CSF60μg/kg。在相应时间点检测脑组织含水量、CD34+血管的表达。结果:治疗组脑组织含水量明显少于ICH组(P<0.05);治疗组CD34+血管表达明显高于ICH组(P<0.05)。结论:脑出血后rhG-CSF通过减轻脑水肿程度、促进血肿周围新生血管生成起到神经组织保护作用。     

重组人粒细胞集落刺激因子对大鼠海马慢性持续性低血流灌注后的修复作用

目的通过对大鼠海马的持续性低血流灌注，观察大鼠海马CA1区锥体细胞、胶质细胞对粒细胞集落刺激因子（G-CSF）动员骨髓干细胞向中枢神经系统迁移的反应，探讨G-CSF对大鼠持续性低血流灌注状态下海马CA1区的病理学修复机制。方法建立大鼠持续性低血流灌注模型（2VO术），术后20周给予重组人粒细胞集落刺激因子（rhG-CSF）动员骨髓干细胞，应用免疫组织化学SP技术和图像分析技术，分析海马CA1区CD34阳性细胞、锥体细胞的数量及其分布状况、以及胶质原纤维酸性蛋白（GFAP）阳性细胞的数量和胞质突起的长度变化。结果连续给予rhG-CSF后大鼠海马CA1区可见CD34阳性细胞存在，提示其动员了骨髓干细胞向中枢神经系统迁移。相对于对照组（生理盐水治疗组），在持续性大脑低灌流状态下，大鼠海马CA1区锥体细胞数量明显增多，GFAP阳性细胞数量明显增多、其胞质突起分支增加、长度明显缩短。结论大鼠海马经持续性低血流灌注后，rhG-CSF可通过动员骨髓细胞向中枢神经系统的迁移，促进海马区锥体细胞、胶质细胞的增生或（和）激活，增强修复能力，为慢性持续性低血流灌注脑损伤的治疗提供了新的思路。     

粒细胞集落刺激因子对实验性脑出血大鼠脑组织微血管重建的影响

目的观察粒细胞集落刺激因子（G-CSF）对脑出血（ICH）大鼠血肿周围CD34^＋血管数的动态变化,探讨G-CSF对ICH大鼠的神经保护作用。方法63只SD大鼠随机分为假手术组、ICH组、治疗组各21只。利用立体定位仪,向SD大鼠右侧苍白球注入断尾获取的自体动脉血制备ICH模型,治疗组于造模1h后腹腔注射重组G-CSF60μg／kg,假手术组、ICH组经腹腔注射等量等渗盐水。三组大鼠于术后6h、24h、48h、72h、7d、14d、21d进行神经功能障碍评分,后处死大鼠,免疫组化检测血肿周围CD34^＋血管数的表达。结果治疗组大鼠从24h开始各时间点神经功能障碍评分较ICH组明显增加（p〈0．05）。假手术组见少许CD34^＋血管表达;ICH组6h开始CD34^＋血管表达增多,14d达高峰;治疗组各时间CD34^＋血管表达较ICH组明显增多（P〈O．05）,高峰提前至7d。结论G-CSF可增加血肿周围新生血管生成,改善神经功能。     

G-CSF对大鼠脑出血后脑水肿及基质金属蛋白酶-9的影响

目的:探讨粒细胞集落刺激因子(G-CSF)对大鼠脑出血(ICH)后脑水肿的影响及可能机制。方法:健康雄性SD大鼠随机分为假手术组、模型组、治疗组。断尾取自体血法制备大鼠ICH模型,治疗组于造模1h后腹腔注射重组粒细胞集落刺激因子(rhG-CSF)60μg/kg。3组大鼠于术后6h、12h、24h、48h、72h、7d处死,干湿重法测定脑组织含水量、免疫组化染色观察MMP-9的动态表达。结果:模型组脑组织含水量明显高于假手术组(P<0.05),在造模后48h、72h最明显,且与6h、12h、24h、7d相比差异显著(P<0.05);治疗组脑组织含水量明显低于模型组(P<0.05)。模型组脑组织MMP-9的表达明显高于假手术组(P<0.05);治疗组各时间点MMP-9的表达低于模型组(P<0.05)。结论:G-CSF可减轻ICH后脑水肿程度,其机制可能与G-CSF抑制MMP-9表达密切相关。     

粒细胞集落刺激因子的神经保护作用及临床试验研究进展

粒细胞集落刺激因子作为一种刺激骨髓粒系造血的生长因子,在临床上被用于治疗各种原因引起的粒细胞减少症。然而,越来越多体内外研究实验及临床试验表明粒细胞集落刺激因子通过激活多种信号转导通路在神经系统损伤性疾病中发挥神经保护作用,包括动员外周干细胞迁移至神经系统、减轻神经细胞凋亡、平衡炎症反应、促进神经干细胞再生及血管生成等,但也存在一定的争议。目前在脑卒中、肌萎缩性侧索硬化及脊髓损伤等神经系统疾病中采用粒细胞集落刺激因子治疗已进入临床试验阶段,在缺氧缺血性脑损伤新生动物模型中也证实其疗效。现将粒细胞集落刺激因子的神经保护作用及临床试验安全性及疗效作一综述。     

红曲素对大鼠脑出血后脑保护作用机制的探讨

目的探讨红曲素(monascin)在脑出血后脑保护作用的机制及最佳剂量。方法 2016年3月—2016年10月于山西医科大学神经电生理实验室选取雄性SD大鼠120只,随机分为假手术组、模型组、低剂量红曲素组、中剂量红曲素组、高剂量红曲素组各24只。采用Ⅳ型胶原酶定位注射制备大鼠脑出血模型,于术后1、3、7、14 d行Garcia神经功能缺损评分,行脑水含量测定、铁染色及PPAR-γ、Nrf2、SRA测定。结果与假手术组相比,模型组、低剂量组、中剂量组及高剂量组神经功能缺损评分在1、3、7、14 d显著降低(F=181.705、494.229、226.576、24.726,P<0.01),且在1、3 d评分最低,脑水含量在1、3 d显著增高(F=21.781、70.036,P<0.05),铁沉积颗粒量在3、7、14 d显著增多(F=21.426、118.122、107.773,P<0.05),且于7~14 d达高峰,PPAR-γ免疫阳性细胞数于1、3、7 d显著增多(F=3.394、28.622、14.12,P<0.05),在3~7 d达高峰,Nrf2免疫阳性细胞数于1、3、7、14 d增多(F=33.076、23.17、47.779、12.182,P<0.05),SRA免疫阳性细胞数于1、3、7、14 d显著增多(F=22.283、511.092、51.811、31.060,P<0.05)。与模型组相比,高剂量组Garcia评分在3~7 d时升高(P<0.05),脑含水量及铁染色颗粒量降低(P<0.05),高剂量组PPAR-γ、Nff2及SPA免疫阳性细胞数于1、3及7 d明显升高(P<0.05),3~7 d达高峰,14 d降至模型组水平。结论高剂量红曲素可增加脑出血后PPAR-γ、Nrf2及SPA阳性细胞数,减轻脑出血后脑水肿及脑铁含量,改善脑出血后神经功能缺损。     

Neuroprotective effect of granulocyte colony-stimulating factor in a focal cerebral ischemic rat model with hyperlipidemia

Granulocyte colony-stimulating factor (G-CSF) has been demonstrated to have neuroprotective effects in rat model with focal cerebral ischemia through anti-apoptotic pathways and by promoting proliferation of neural stem cells. In the present study, we examined the neuroprotective effect of G-CSF in an acute focal cerebral ischemia rat model with lipid metabolism disorder. Eighty male SD rats were randomly divided into normal diet control group (NC group) and high-fat diet group (HFD group) (n = 40 in each). In HFD group, rats were fed on high fat diet to induce atherosclerosis. After 29 days, 4 rats from each group were sacrificed to evaluate the effects of different diets, and the middle cerebral artery occlusion (MCAO) was performed in the rest of the rats. MCAO rats received either G-CSF (50 μg·kg-1·mL-1) or phosphate buffered saline (PBS) injection through the external jugular vein for 5 days, which was followed by 5-bromo-deoxy uridine (BrdU, i.p., 50 mg/kg) injection for another 7 days. To evaluate the effects of G-CSF treatment on neurological function, the modified neurological severity score (mNSS) was calculated. The vascular distribution, ischemic cells proliferation, cell apoptosis and the expression of vascular endothelial growth factor (VEGF) were measured to determine the effects of G-CSF treatment. Our results showed that G-CSF-treated rats had a lower mNSS than PBS-treated rats in both NC group and HFD group. G-CSF injection promoted endothelial cell proliferation and vascular regeneration, and inhibited cell apoptosis. The serum and tissue levels of VEGF were significantly increased after G-CSF treatment. It is concluded that G-CSF exerts its neuroprotective effect in focal cerebral ischemia rats with hyperlipidemia by enhancing angiogenesis, promoting cells proliferation, decreasing cell apoptosis, and increasing local VEGF expression.     

Neuroprotective effects of edaravone on early brain injury in rats after subarachnoid hemorrhage

Background The underlying mechanism of early neurobiological impairment after subarachnoid hemorrhage （SAH） is not well understood, but the system of reactive oxygen superoxide （ROS） might be involved. Edaravone （MCI-186）, a potent free radical scavenger that prevents apoptosis of neurons, was thus used in this study to see its possible therapeutic effect in early brain injury due to SAH in a rat model. Methods One hundred and twenty male Sprague-Dawley rats were randomly assigned to four groups： group 1, control rats receiving sham operation only; group 2, rats with SAH treated by saline; group 3, rats with SAH treated with 1 mg/kg MCI-186 injected intraperitoneally; and group 4, rats with SAH treated with 3 mg/kg MCI-186. Treated with either saline or MCI-186 twice daily for two consecutive days after SAH, the rats were sacrificed for measurements of malondialdehyde （MDA） and activity of superoxide dismutase （SOD） and histological analysis of caspase-3 protein by Western blotting and immunohistochemical staining. In addition, mortality and neurological scores were statistically analyzed by the chi-square test and Dunn＇s procedure respectively for each group. One-way analysis of variance followed by the Tukey＇s procedure was also used in data analysis. Results The rats in group 2 that received saline only showed neurological impairment as well as elevated mortality, and were found to have significantly increased levels of MDA and caspase-3, but reduced SOD activities in brain tissues （P 〈0.05）. When treated with MCI-186 at two different dosages, the rats in groups 3 and 4 had markedly decreased levels of MDA and caspase-3 but increased SOD activities in the brain tissue （P 〈0.05）, along with improved scores of neurological evaluation （P 〈0.05）. Conclusions This study sheds some lights on the therapy of SAH-induced early brain injury by providing the promising data indicating that MCI-186, a radical scavenger, can efficiently diminish apoptosis of neurons and thus prevent the function loss of the brain in rats with SAH.     

老年大鼠脑出血后神经细胞再生的研究

目的：探讨老年大鼠脑出血后发生的神经细胞再生现象及其特点。方法：应用成年及老年（分别3个月及18个月大小）雄性Sprague—Dawley大鼠制作脑出血模型，主要采用蛋白印迹分析、免疫组织化学染色等方法，对Bromodeoxyuridine（Brdu）及未成熟神经元标记doublecortin（DCX）和polysialylated neural cell adhesion protein（PSA—NCAM）等进行分析。结杲：在老年大鼠，DCX的表达于脑出血后7天即有增加，2周后同侧脑室下区（subventricular zone，SVZ）及基底节可见大量DCX阳性细胞，其表达达到高峰，是对照组的（670&#177;145）％（P〈0．01）；绝大多数的DCX阳性的细胞也共同表达Brdu；与成年大鼠相比，老年大鼠脑出血后DCX表达减少。结论：老年大鼠脑出血后有明显的神经细胞再生，SVZ的神经元前体细胞增生明显，并向受损伤的基底节区移行。     

Mobilization efficiency of granulocyte colony-stimulating factor and stem cell factor to bone marrow mononuclear cells and mechanisms

The mobilization efficiency of granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) to bone marrow mononuclear cells (MNCs) in mice was observed, and the changes of CXCL12/CXCR4 signal were detected in order to find out the mobilization mechanism of stem cells. Kunming mice were randomly divided into two groups. The mice in treatment group were subjected to subcutaneous injection of G-CSF at a dose of 100 μg/kg and SCF at a dose of 25 μg/kg every day for 5 days, and those in control grou...     

粒细胞集落刺激因子治疗大鼠局灶性脑缺血再灌注损伤

OBJECTIVE: To study the therapeutic effects of granulocyte colony-stimulating factor (G-CSF) on focal cerebral ischemia-reperfusion injury in rats. METHODS: Adult male SD rats were subjected to transient (2 h) middle cerebral artery occlusion (MCAO) and scored 48 h later for such neurological functions as spontaneous movement, paresis, forelimb motor function, climbing ability, pain sensation, and position sense. Twenty rats with the total score less than 12 were divided equally into two groups to receive daily subcutaneous injection of 20 microg/kg of G-CSF for 19 days or the same dose of saline injection (control group). The rats in the two groups were also given intraperitoneal injection of Brdu10 mg/kg for 19 days. The neurological functions of the rats in both groups were examined on a weekly basis after MCAO and on day 21, the rats were killed to prepare frozen sections of the brain tissues for double immunohistochemical staining with Brdu and glial fibrillary acidic protein antibody to identify the neural cells newly evolved from the stem cells. RESULTS: The rats in G-CSF group showed better neurological function recovery than the control rats 3 weeks after MCAO (P<0.05). Obvious regeneration of the neural cells was observed around the infarction area in the rats receiving G-CSF treatment. CONCLUSION: G-CSF promotes neurological function recovery and neural cell regeneration after cerebral infarction in rats and can be effective for intervention of focal cerebral ischemia-reperfusion injury.     

粒细胞集落刺激因子治疗大鼠局灶性脑缺血再灌注损伤

目的 探讨粒细胞集落刺激因子(G-CSF)对大鼠局灶性脑缺血再灌注损伤的治疗作用.方法 线栓法构建SD大鼠脑缺血再灌注损伤模型,线栓时间2 h.手术48 h后,对模型鼠神经功能缺失评分,将12分以下的20只大鼠随机平均分为两组:干预性治疗组:予G-CSF 20μg/(kg·d)皮下注射,共19 d;对照组皮下注射等量生理盐水,置同样环境饲养.两组同时腹膜腔注射脱氧溴化尿嘧啶(Brdu)10mg/kg共19 d.两组均于手术后每周进行神经系统功能缺失评测.术后3周,将实验鼠麻醉后处死取脑,脑沿冠状面将鼠脑切成等分,取第3块脑切块,做冰冻切片,行神经干细胞及神经胶质细胞特异性抗体与Brdu抗体免疫组化双染,观察梗死区新生神经细胞.结果 手术后3周,与对照组比较,G-CSF治疗组神经功能恢复好于对照组(P<0.0);治疗组脑组织梗塞灶周围新生神经干细胞和神经胶质细胞增生明显多于对照组.结论 G-CSF能促进脑梗死后神经功能恢复和神经细胞再生,是一有效的脑缺血干预性治疗用药.     

Role of granulocyte colony-stimulating factor in paclitaxel-induced intestinal barrier breakdown and bacterial translocation in rats

Background  Chemotherapy causes breakdown of the intestinal barrier, which may lead to bacterial translocation. Paclitaxel, an anti-tubulin agent, has many side effects; however, its effect on the intestinal barrier is unknown. Previous studies show that granulocyte colony-stimulating factor (G-CSF) plays an important role in modulating intestinal barrier function, but these studies are not conclusive. Here, we investigated the effects of paclitaxel on the intestinal barrier, and whether G-CSF could prevent paclitaxel-induced bacterial translocation.

Methods  Twenty-four male Sprague-Dawley rats were divided into three groups: control group, paclitaxel group and paclitaxel + G-CSF group. Intestinal permeability was measured by the urinary excretion rates of lactulose and mannitol administered by gavage. The mesenteric lymph nodes, spleen and liver were aseptically harvested for bacterial culture. Endotoxin levels and white blood cell (WBC) counts were measured and bacterial quantification performed using relative real-time PCR. Jejunum samples were also obtained for histological observation. Intestinal apoptosis was evaluated using a fragmented DNA assay and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP)-biotin nick end-labeling staining. One-way analysis of variance and Fisher’s exact test were used to compare differences between groups.

Results  Paclitaxel induced apoptosis in 12.5% of jejunum villus cells, which was reduced to 3.8% by G-CSF treatment. Apoptosis in the control group was 0.6%. Paclitaxel treatment also resulted in villus atrophy, increased intestinal permeability and a reduction in the WBC count. G-CSF treatment resulted in increased villus height and returned WBC counts to normal levels. No bacterial translocation was detected in the control group, whereas 6/8, 8/8, and 8/8 rats in the paclitaxel group were culture-positive in the liver, spleen and mesenteric lymph nodes, respectively. Bacterial translocation was partially inhibited by G-CSF.

Conclusions  Paclitaxel disrupts the intestinal barrier, resulting in bacterial translocation. G-CSF treatment protects the intestinal barrier, prevents bacterial translocation, and attenuates paclitaxel-induced intestinal side-effects.