CAJ | 학술논문

目的探讨创面愈合过程中粒-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)的表达变化及其在创面愈合中的作用和机制.方法采用小鼠全层皮肤缺损伤模型.肌肉麻醉后在背部中线近颈侧制作1.0 cm×1.0 cm大小皮肤缺损伤创而.50只雄性小鼠创面造模成功后,单笼饲养,将每笼编号,依次为1～50号,按完全随机原则将小鼠分为对照组(25只)和GM-CSF治疗组(25只),每组分为5个时相点,分别为5只.治疗组创面用重组GM-CSF(rhGM-CSF)凝胶(10μg/cm2),对照组创面用凝胶基质.于伤后第3,5,7,10和14天,观察创面愈合时间;测定创而愈合率;切取创面组织,检测病理组织学变化;通过CD31免疫组化染色结果分析,计算创面微血管密度;应用RTvPCR检测创面GM-CSF、血小板源性生长因子(platelet-derived growth factor,PDGF)、血管内皮生长因子(vascular endothelial growth factor,VEGF)和基质细胞衍生因子-1(stromal cell derived factor-1,SDF-1)基因表达变化.结果 RT-PCR检测结果显示创面GM-CSF基因表达伤后3 d达峰值(P＜0.01),直到伤后10 d均维持较高水平(P＜0.05),伤后14 d其表达显著下降接近正常;应用rhGM-CSF凝胶后,创面愈合时间较对照组提前(2.4±0.3)d,其创而愈合率于伤后7～14 d显著升高(P＜0.05);组织学显示创伤早期创面中性粒细胞数量较少,创沿上皮细胞增殖数量较多,创面肉芽组织增生明显,并且细胞密度大,以梭形细胞和卵圆形细胞为主,新生血管数量较多;创面微血管密度在伤后7～14 d显著增加(P＜0.05);VEGF和SDF-1基因表达,分别于伤后7 d和10 d内显著上调表达(P＜0.05),促愈生长因子PDGF基因于伤后各时相点均显著上调表达(P＜0.05).结论 GM-CSF在创面愈合早期表达增高,GM-CSF可促进创而愈合,其作用机制与促进创面促血管生成因子和促愈合生长因子基因表达上凋有关.
목적 탐토창면유합과정중립-거서세포집락자격인자(granulocyte-macrophage colony stimulating factor,GM-CSF)적표체변화급기재창면유합중적작용화궤제.방법채용소서전층피부결손상모형.기육마취후재배부중선근경측제작1.0 cm×1.0 cm대소피부결손상창이.50지웅성소서창면조모성공후,단롱사양,장매롱편호,의차위1～50호,안완전수궤원칙장소서분위대조조(25지)화GM-CSF치료조(25지),매조분위5개시상점,분별위5지.치료조창면용중조GM-CSF(rhGM-CSF)응효(10μg/cm2),대조조창면용응효기질.우상후제3,5,7,10화14천,관찰창면유합시간;측정창이유합솔;절취창면조직,검측병리조직학변화;통과CD31면역조화염색결과분석,계산창면미혈관밀도;응용RTvPCR검측창면GM-CSF、혈소판원성생장인자(platelet-derived growth factor,PDGF)、혈관내피생장인자(vascular endothelial growth factor,VEGF)화기질세포연생인자-1(stromal cell derived factor-1,SDF-1)기인표체변화.결과 RT-PCR검측결과현시창면GM-CSF기인표체상후3 d체봉치(P＜0.01),직도상후10 d균유지교고수평(P＜0.05),상후14 d기표체현저하강접근정상;응용rhGM-CSF응효후,창면유합시간교대조조제전(2.4±0.3)d,기창이유합솔우상후7～14 d현저승고(P＜0.05);조직학현시창상조기창면중성립세포수량교소,창연상피세포증식수량교다,창면육아조직증생명현,병차세포밀도대,이사형세포화란원형세포위주,신생혈관수량교다;창면미혈관밀도재상후7～14 d현저증가(P＜0.05);VEGF화SDF-1기인표체,분별우상후7 d화10 d내현저상조표체(P＜0.05),촉유생장인자PDGF기인우상후각시상점균현저상조표체(P＜0.05).결론 GM-CSF재창면유합조기표체증고,GM-CSF가촉진창이유합,기작용궤제여촉진창면촉혈관생성인자화촉유합생장인자기인표체상조유관.
Objective To investigate the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) and its associated mechanism during the wound healing. Methods The animal model with the full-thickness skin injury was used in the study. Fifty male mice were involved in the study and divided randomly into control group (n = 25) and GM-CSF group (n = 25). Each group had five time points (5 mice per time point). All the mice received full-thickness skin defect (1 cm × 1 cm) through the panniculus camosus on the midline of the back near the neck after anesthesia. Recombinant human granulocyte-macrophage colony stimulating factor (RhGM-CSF) gel (10 μg/cm2) were applied in the GM-CSF group and gel matrix without RhGM-CSF applied in the control group. The wound healing time and rate were observed at days 3, 5, 7, 10 and 14 after injury. The wound specimens were collected to detect the histopathological change. The microvessel density of the wound was counted based on the results of CD31 immunohistochemistry. RT-PCR was employed to detect the expression changes of GMCSF, platelet-derived growth factor (PDGF) , vascular endothelial growth factor ( VEGF) and stromal cell derived factor-1 (SDF-1). Results RT-PCR results showed that the gene expression of GM-CSF reached the peak at day 3 after injury (P＜0. 01) and kept the high level at days 3-10 after injury (P＜ 0. 05) , followed by a sharp decrease to a normal level at day 14 after wound. The wound healing time was average (2.4 ±0. 3) days earlier than the control mice after application of rhGM-CSF, with significant increase of the wound healing rate during 7-14 days after injury ( P ＜ 0. 05 ). In the GM-CSF group, the early histology of trauma wound showed a small number of neutrophils, obvious epithelial cell proliferation in the wound margin, marked hyperplasia of the granulation tissue, high cell density with quantity of spindle-shaped and oval-shaped cells and increased number of new blood vessels. The microvessel density was also increased significantly (P ＜ 0. 05) at days 7-14 after injury. The gene expressions of VEGF and SDF-1 were significantly increased at day 7 and day 10 respectively after injury (P＜0.05) and the gene expression of pro-healing factor PDGF was significantly increased in every time point (at days 5, 7 and 10,P＜0.05;at day 14,P＜0.01). Conclusion GM-CSF expresses highly in the early stage after injury and can promote the wound healing, when the mechanism may relate to the up-regulated expressions of pro-angiogenic factors and pro-healing growth factors.