中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
4期
697-700
,共4页
赵菁玲%徐盈斌%唐锦明%舒斌%薛亮%谢举临%祁少海%刘旭盛
趙菁玲%徐盈斌%唐錦明%舒斌%薛亮%謝舉臨%祁少海%劉旭盛
조정령%서영빈%당금명%서빈%설량%사거림%기소해%류욱성
粒细胞-巨噬细胞集落刺激因子%血管生成素%烧伤%创面愈合
粒細胞-巨噬細胞集落刺激因子%血管生成素%燒傷%創麵愈閤
립세포-거서세포집락자격인자%혈관생성소%소상%창면유합
Granulocyte macrophage colony stimulating factor%Angiopoietin%Burn%Wound healing
目的 观察粒细胞-巨噬细胞集落刺激因子(GM-CSF)对大鼠深Ⅱ度烫伤创面愈合过程中血管生成素-1(Ang-1)表达的影响.方法 建立SD大鼠深Ⅱ度烫伤模型,按自身对照的方法将创面随机分为GM-CSF实验组和对照组.分别取伤后1、3、7、14、21 d创面皮肤组织,采用实时荧光定量逆转录-聚合酶链反应(FQ-PCR)分析Ang-1的mRNA表达；免疫组织化学法和Western blot法检测创面Ang-1蛋白的表达；免疫组织化学法检测创面CD31的表达,计算微血管密度(MVD).结果 创面Ang-1的mRNA表达量于伤后1、3d呈下降趋势,实验组(1.44±0.20、0.30 ±0.17)低于对照组(2.36±0.18、1.34±0.24) (P ＜0.05)；之后其表达量逐渐升高,伤后7、14、21 d实验组(4.52±0.21、8.47±0.14、7.52±0.20)均明显高于对照组(2.71±0.12、3.69±0.24、3.07±0.17)(P＜0.05).实验组创面Ang-1的蛋白表达量于伤后1、3d可见显著下降(0.491±0.055、0.181±0.025),与对照组(0.735±0.080、0.436±0.071)比较明显降低(P＜0.05)；之后其表达量持续升高,至伤后14 d达峰值,实验组(0.821±0.060、1.257±0.225、1.126±0.131)较对照组(0.701±0.078、0.825±0.072、0.733±0.073)明显增高(P＜0.05).实验组创面于伤后第3天起见较多CD31阳性血管并逐渐增多,MVD值在伤后3、7、14、21 d(9.5±1.8、23.4±2.9、38.4±3.1、24.6±2.5)均明显高于对照组(4.0±1.9、10.5±2.0、22.9±2.7、15.1±2.6,P＜0.05).结论 GM-CSF降低Ang-1在烧伤创面愈合早期的表达,有利于创面血管新生；促进Ang-1在愈合中、后期表达,利于创面血管成熟,加快创面愈合.
目的 觀察粒細胞-巨噬細胞集落刺激因子(GM-CSF)對大鼠深Ⅱ度燙傷創麵愈閤過程中血管生成素-1(Ang-1)錶達的影響.方法 建立SD大鼠深Ⅱ度燙傷模型,按自身對照的方法將創麵隨機分為GM-CSF實驗組和對照組.分彆取傷後1、3、7、14、21 d創麵皮膚組織,採用實時熒光定量逆轉錄-聚閤酶鏈反應(FQ-PCR)分析Ang-1的mRNA錶達；免疫組織化學法和Western blot法檢測創麵Ang-1蛋白的錶達；免疫組織化學法檢測創麵CD31的錶達,計算微血管密度(MVD).結果 創麵Ang-1的mRNA錶達量于傷後1、3d呈下降趨勢,實驗組(1.44±0.20、0.30 ±0.17)低于對照組(2.36±0.18、1.34±0.24) (P ＜0.05)；之後其錶達量逐漸升高,傷後7、14、21 d實驗組(4.52±0.21、8.47±0.14、7.52±0.20)均明顯高于對照組(2.71±0.12、3.69±0.24、3.07±0.17)(P＜0.05).實驗組創麵Ang-1的蛋白錶達量于傷後1、3d可見顯著下降(0.491±0.055、0.181±0.025),與對照組(0.735±0.080、0.436±0.071)比較明顯降低(P＜0.05)；之後其錶達量持續升高,至傷後14 d達峰值,實驗組(0.821±0.060、1.257±0.225、1.126±0.131)較對照組(0.701±0.078、0.825±0.072、0.733±0.073)明顯增高(P＜0.05).實驗組創麵于傷後第3天起見較多CD31暘性血管併逐漸增多,MVD值在傷後3、7、14、21 d(9.5±1.8、23.4±2.9、38.4±3.1、24.6±2.5)均明顯高于對照組(4.0±1.9、10.5±2.0、22.9±2.7、15.1±2.6,P＜0.05).結論 GM-CSF降低Ang-1在燒傷創麵愈閤早期的錶達,有利于創麵血管新生；促進Ang-1在愈閤中、後期錶達,利于創麵血管成熟,加快創麵愈閤.
목적 관찰립세포-거서세포집락자격인자(GM-CSF)대대서심Ⅱ도탕상창면유합과정중혈관생성소-1(Ang-1)표체적영향.방법 건립SD대서심Ⅱ도탕상모형,안자신대조적방법장창면수궤분위GM-CSF실험조화대조조.분별취상후1、3、7、14、21 d창면피부조직,채용실시형광정량역전록-취합매련반응(FQ-PCR)분석Ang-1적mRNA표체；면역조직화학법화Western blot법검측창면Ang-1단백적표체；면역조직화학법검측창면CD31적표체,계산미혈관밀도(MVD).결과 창면Ang-1적mRNA표체량우상후1、3d정하강추세,실험조(1.44±0.20、0.30 ±0.17)저우대조조(2.36±0.18、1.34±0.24) (P ＜0.05)；지후기표체량축점승고,상후7、14、21 d실험조(4.52±0.21、8.47±0.14、7.52±0.20)균명현고우대조조(2.71±0.12、3.69±0.24、3.07±0.17)(P＜0.05).실험조창면Ang-1적단백표체량우상후1、3d가견현저하강(0.491±0.055、0.181±0.025),여대조조(0.735±0.080、0.436±0.071)비교명현강저(P＜0.05)；지후기표체량지속승고,지상후14 d체봉치,실험조(0.821±0.060、1.257±0.225、1.126±0.131)교대조조(0.701±0.078、0.825±0.072、0.733±0.073)명현증고(P＜0.05).실험조창면우상후제3천기견교다CD31양성혈관병축점증다,MVD치재상후3、7、14、21 d(9.5±1.8、23.4±2.9、38.4±3.1、24.6±2.5)균명현고우대조조(4.0±1.9、10.5±2.0、22.9±2.7、15.1±2.6,P＜0.05).결론 GM-CSF강저Ang-1재소상창면유합조기적표체,유리우창면혈관신생；촉진Ang-1재유합중、후기표체,리우창면혈관성숙,가쾌창면유합.
Objective To investigate the influence of granulocyte macrophage colony stimulating factor (GM-CSF) on the expression of angiopoietin-1 (Ang-1) in deep partial thickness burn wound in rats.Methods The models of deep partial thickness burn wounds on the back in Spregue-Dawley (SD) rats were established.The paired wounds on the same rats were randomly divided into GM-CSF treatment (test) and control groups by self-control.Tissue samples were harvested from the wounds on the 1 st,3rd,7th,14th and 21st day respectively after burns.The mRNA expression of Ang-1 was detected by using real-time reverse transcriptase-polymerase chain reaction (FQ-PCR).And the protein expression of Ang-1 was detected by immunohistochemical and western blot,the protein expression of CD31 by using immunohistochemical staining,and the microvascular density was calculated.Results The mRNA expression level of Ang-1 in the burn wounds was decreased slightly on the 1st and 3rd postburn days and that in test group (1.44 ±0.20,0.30 ± 0.17) was lower than that in control group (2.36 ± 0.18,1.34 ± 0.24) (P ＜0.05).Then the Ang-1 mRNA expression was gradually increased on the 7th and 14th postburn days and decreased slightly on the 21st postburn day,and the expression level in the test group at the above time points (4.52 ±0.21,8.47 ±0.14,7.52 ±0.20) was higher than that in control group (2.71 ±0.12,3.69 ±0.24,3.07 ±0.17) (P ＜0.05).The protein expression of Ang-1 was declined remarkably on the 1st and 3rd postburn days,which in test group (0.491 ±0.055,0.181 ±0.025) was lower than that in control group (0.735 ±0.080,0.436 ±0.071) (P ＜0.05).Then the Ang-1 protein expression was gradually increased and reached the peak on the 14th postburn day and the expression in the test group (0.821 ±0.060,1.257 ± 0.225,1.126 ± 0.131) was higher than that in the control group (0.701 ± 0.078,0.825 ± 0.072,0.733 ±0.073) (P ＜0.05).The microvascular number with positive CD31 expression was increased evidently from the 3rd postburn day in test group,and the counting of MVD was remarkably higher in the test group (9.5±1.8,23.4±2.9,38.4±3.1,24.6±2.5) than in the control group (4.0±1.9,10.5±2.0,22.9 ±2.7,15.1 ±2.6) on the 3rd,7th,14th and 21st days respectively after burns (P ＜0.05).Conclusion GM-CSF inhibits the mRNA and protein expression of Ang-1 during the early stage of wound healing,which can be beneficial to the angiogenesis.And GM-CSF boosts Ang-1 expression during the middle and late stages of wound healing.This would be helpful to wound vascular maturation and wound healing.
