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重组鼠粒细胞-巨噬细胞集落刺激因子对高氧暴露新生大鼠肺损伤的影响

Effects of recombinant murine granulocyte macrophage-colony-stimulating factor on lung injury of hyperoxia-exposed newborn rats

摘要:

目的 探讨重组鼠粒细胞-巨噬细胞集落刺激因子(GM-CSF)对高氧暴露新生大鼠肺损伤的影响.方法 32只3日龄新生大鼠随机分为高氧+GM-CSF组、空气+GM-CSF组、高氧对照组和空气对照组,每组8只.高氧组新生大鼠均采用95%的氧浓度暴露7 d,空气组仅置于空气环境中;高氧+GM-CSF组和空气+GM-CSF组大鼠分别于实验的24、72和120 h皮下注射GM-CSF,每次9 μg/kg,高氧对照组和空气对照组于相同时点皮下注射相同体积生理盐水.支气管肺泡灌洗液细胞计数检测白细胞并进行细胞分类计数;ELISA法检测肺组织匀浆TGF-β1、TNF-α水平;HE染色检测肺组织辐射状肺泡计数(RAC)值;免疫组织化学方法检测肺组织端粒酶逆转录酶(TERT)积分和细胞核增殖抗原(PCNA)指数.结果 (1)高氧+GM-CSF组、高氧对照组支气管肺泡灌洗液中自细胞总数、分叶核细胞及巨噬细胞数均明显高于空气+GM-CSF组和空气对照组(P均<0.05).(2)与空气对照组相比,高氧+GM-CSF组、高氧对照组RAC[分别为(11.15±1.33)个和(9.59±0.67)个]显著降低,TNF-α[分别为(142.93±24.02)pg/ml和(224.09±41.91)pg/ml]和TGF-β1水平[分别为(1726.48±91.09)pg/ml和(2047.72±152.01)pg/ml]显著升高(P<0.05).(3)四组肺组织中均有PCNA阳性表达,绝大多数定位于气道和肺泡上皮细胞,表现为核染蓝色.与空气对照组相比,高氧+GM-CSF组、高氧对照组PCNA表达明显下降(P均<0.05),而高氧+GM-CSF组表达高于高氧对照组(P<0.05).TERT阳性显色部位在胞核,主要分布于外周肺组织肺间隔和肺泡壁部位,呈散在分布.与其他三组比较,高氧+GM-CSF组TERT表达明显增高(P均<0.05).结论 GM-CSF对新生大鼠高氧肺损伤具有保护作用,可能与增加肺组织干细胞的增殖及改善局部微环境等有关.

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abstracts:

Objective To investigate the effects of systemic treatment with recombinant murine granulocyte macrophage-colony stimulating factor (GM-CSF) on lung injury in hyperoxia-exposed newborn rats. Methods Thirty-two 3-day-old SD rats were randomly divided into four groups: hyperoxia exposed + GM-CSF group (Group 1),air-exposed+ GM-CSF group (Group 2), hyperoxia exposed group (Group 3) and air-exposed group (Group 4). The rats in Group 1 and 3 were placed in a sealed Plexiglas chamber maintaining O2 levels above 95% and those in Group 2 and 4 were exposed to air. Rats in Group 1 and 2 were injected with GM-CSF (9 μg/kg ih. ) at 24 h, 72 h and 120 h, respectively and those in Group 3 and 4 received subcutaneous injection of saline at the same time point. All rats were sacrificed 7 d later, and immunohistochemistry was used to evaluate the expression of telomerase reverse transcriptase (TERT) grade and proliferating cell nuclear antigen (PCNA) index in lung tissue; and total numbers of cells in bronchoalveolar lavage fluid (BALF) were counted and the percentages of mononuclear cells and neutrophils were calculated. The levels of TNFα and TGFβ1 of tissue homogenates were detected by ELISA. The value of radical alveolar count(RAC) was calculated. Results (1) There were significant differences in the values of RAC, total numbers of white blood cells (WBC) and the percentages of mononuclear cells and neutrophils in BALF, the levels of TNFα and TGFβ1 of tissue homogenates and PCNA index among the four groups. (2) Compared with Group 4, both PCNA index and RAC decreased (P<0.05), and the levels of TNFα and TGFβ1, WBC, mononuclear cells and neutrophil count increased in Group 3. Compared to Group 3, both PCNA index and RAC increased (P<0.05), and the levels of TNFα and TGFβ1 decreased in Group 1 (P<0. 05), but no differences was found in the WBC, mononuclear cells and neutrophil count (P>0. 05). (3) TERT grade in Group 1 was obviously increased, while there were no difference among the other groups. Conclusions The protective effect of GM-CSF on lung injury of newborn rats suffered from hyperoxia may be related to the increase of stem cells proliferation and the improvement of local microenvironment.

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作者: 田兆方 [1] 李玉红 [1] 付雪梅 [2] 封志纯 [2] 杜江 [3]
期刊: 《中华围产医学杂志》2009年12卷3期 177-181页 ISTICPKUCSCDCA
分类号: R71
栏目名称: 高氧暴露与新生儿肺损伤
DOI: 10.3760/cma.j.issn.1007-9408.2009.03.006
发布时间: 2009-06-26
基金项目:
国家自然科学基金面上项目 南京医科大学科技发展基金重点项目
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