目的 探讨平阳霉素对婴幼儿血管瘤的作用机制.方法 取24只裸鼠建立婴幼儿血管瘤模型,将裸鼠分为实验组和空白对照组,各12只.将平阳霉素注入实验组血管瘤内,观察其形态变化,采用普通光学和电子显微镜观察血管瘤组织结构的变化,然后利用cDNA基因芯片检测血管瘤组织基因表达谱的变化.结果 血管瘤注药后体积逐渐缩小变硬,充血程度减轻,1个月后血管瘤体基本消失.光镜下观察见血管瘤失去组织结构,大部分细胞溶解坏死,大片的胶原纤维取而代之.电镜下观察见细胞大片溶解,未见完整的细胞结构及血管,可见凋亡细胞和凋亡小体.基因芯片研究结果表明,与空白对照组比较共有33个基因表达差异在2倍以上,其中9个与凋亡有关的基因,分别是鼠双微体2 (murine double minute 2,MDM2)、不耐热肠毒素B亚单位(heat-labile enterotoxin B subunit,LTB)、淋巴毒素B受体(lymphotoxin B receptor LTBR)、肿瘤坏死因子超家族7(tumor necrosis factor ligand superfamily 7,TNFSF7)、肿瘤坏死因子受体超家族21(tumor necrosis factor receptor superfamily 21,TNFRSF21)、TNFRSFlA、人髓细胞白血病基因l(myeloid cell leukemia-1,MCL1)和半胱天冬酶3(caspase3,CASP3).13个与细胞增殖和细胞周期相关基因,分别是细胞分裂周期蛋白27(cell division cycle27,CDC27)、CDC37、CDC28蛋白激酶1B(CDC28 protein kinase 1B,CKS1B)、细胞周期蛋白B1 (cycling B1,CCNB1)、CUL2(cullin 2)、CUL3(cullin 3)、CUL4A(cullin 4A)、生长抑制和DNA损伤诱导基因45A (growth arrest and DNA damage-inducible 45A,Gadd45A)、减数分裂重组11同源体B(meiotic recombination 11 homolog B,MRE11 B)、叉头框M1 (forkhead box M1,FOXM1)、微型染色体7(minichromosome maintenance 7,MCM7)、单克隆抗体Ki-67抗原(antigen identified by monoclonal antibody Ki67,MKI67)和细胞增殖核抗原(proliferating cell nuclear antigen,PCNA).11个与细胞应激和毒性反应有关的基因分别是谷胱甘肽过氧化物酶1(glutathione peroxidase 1,GPX1)、金属硫蛋白1A(metallothionein,MT1A)、超氧化物歧化酶1(superoxide dismutase-1,SOD1)、热休克蛋白A1A(heatshock protein A1A,HSPA1A)、HSPA2、HSPA4、HSPA5、HSP9B、HSPCA、巨噬细胞游走抑制因子(macrophage migration inhibitory factor,MIF)和纤溶酶原激活物抑制因子1(plasminogen activator inhibitor,PAI1).结论 平阳霉素治疗婴幼儿血管瘤是多因素、多水平综合作用的结果,平阳霉素能够作用于细胞DNA的多个片段,既能诱发细胞凋亡,抑制细胞的增殖,还能直接使细胞的应激与毒性反应能力降低,细胞毒性作用和诱导细胞凋亡是同一作用机制的两种表现形式,并非两种作用机制.
目的 探討平暘黴素對嬰幼兒血管瘤的作用機製.方法 取24隻裸鼠建立嬰幼兒血管瘤模型,將裸鼠分為實驗組和空白對照組,各12隻.將平暘黴素註入實驗組血管瘤內,觀察其形態變化,採用普通光學和電子顯微鏡觀察血管瘤組織結構的變化,然後利用cDNA基因芯片檢測血管瘤組織基因錶達譜的變化.結果 血管瘤註藥後體積逐漸縮小變硬,充血程度減輕,1箇月後血管瘤體基本消失.光鏡下觀察見血管瘤失去組織結構,大部分細胞溶解壞死,大片的膠原纖維取而代之.電鏡下觀察見細胞大片溶解,未見完整的細胞結構及血管,可見凋亡細胞和凋亡小體.基因芯片研究結果錶明,與空白對照組比較共有33箇基因錶達差異在2倍以上,其中9箇與凋亡有關的基因,分彆是鼠雙微體2 (murine double minute 2,MDM2)、不耐熱腸毒素B亞單位(heat-labile enterotoxin B subunit,LTB)、淋巴毒素B受體(lymphotoxin B receptor LTBR)、腫瘤壞死因子超傢族7(tumor necrosis factor ligand superfamily 7,TNFSF7)、腫瘤壞死因子受體超傢族21(tumor necrosis factor receptor superfamily 21,TNFRSF21)、TNFRSFlA、人髓細胞白血病基因l(myeloid cell leukemia-1,MCL1)和半胱天鼕酶3(caspase3,CASP3).13箇與細胞增殖和細胞週期相關基因,分彆是細胞分裂週期蛋白27(cell division cycle27,CDC27)、CDC37、CDC28蛋白激酶1B(CDC28 protein kinase 1B,CKS1B)、細胞週期蛋白B1 (cycling B1,CCNB1)、CUL2(cullin 2)、CUL3(cullin 3)、CUL4A(cullin 4A)、生長抑製和DNA損傷誘導基因45A (growth arrest and DNA damage-inducible 45A,Gadd45A)、減數分裂重組11同源體B(meiotic recombination 11 homolog B,MRE11 B)、扠頭框M1 (forkhead box M1,FOXM1)、微型染色體7(minichromosome maintenance 7,MCM7)、單剋隆抗體Ki-67抗原(antigen identified by monoclonal antibody Ki67,MKI67)和細胞增殖覈抗原(proliferating cell nuclear antigen,PCNA).11箇與細胞應激和毒性反應有關的基因分彆是穀胱甘肽過氧化物酶1(glutathione peroxidase 1,GPX1)、金屬硫蛋白1A(metallothionein,MT1A)、超氧化物歧化酶1(superoxide dismutase-1,SOD1)、熱休剋蛋白A1A(heatshock protein A1A,HSPA1A)、HSPA2、HSPA4、HSPA5、HSP9B、HSPCA、巨噬細胞遊走抑製因子(macrophage migration inhibitory factor,MIF)和纖溶酶原激活物抑製因子1(plasminogen activator inhibitor,PAI1).結論 平暘黴素治療嬰幼兒血管瘤是多因素、多水平綜閤作用的結果,平暘黴素能夠作用于細胞DNA的多箇片段,既能誘髮細胞凋亡,抑製細胞的增殖,還能直接使細胞的應激與毒性反應能力降低,細胞毒性作用和誘導細胞凋亡是同一作用機製的兩種錶現形式,併非兩種作用機製.
목적 탐토평양매소대영유인혈관류적작용궤제.방법 취24지라서건립영유인혈관류모형,장라서분위실험조화공백대조조,각12지.장평양매소주입실험조혈관류내,관찰기형태변화,채용보통광학화전자현미경관찰혈관류조직결구적변화,연후이용cDNA기인심편검측혈관류조직기인표체보적변화.결과 혈관류주약후체적축점축소변경,충혈정도감경,1개월후혈관류체기본소실.광경하관찰견혈관류실거조직결구,대부분세포용해배사,대편적효원섬유취이대지.전경하관찰견세포대편용해,미견완정적세포결구급혈관,가견조망세포화조망소체.기인심편연구결과표명,여공백대조조비교공유33개기인표체차이재2배이상,기중9개여조망유관적기인,분별시서쌍미체2 (murine double minute 2,MDM2)、불내열장독소B아단위(heat-labile enterotoxin B subunit,LTB)、림파독소B수체(lymphotoxin B receptor LTBR)、종류배사인자초가족7(tumor necrosis factor ligand superfamily 7,TNFSF7)、종류배사인자수체초가족21(tumor necrosis factor receptor superfamily 21,TNFRSF21)、TNFRSFlA、인수세포백혈병기인l(myeloid cell leukemia-1,MCL1)화반광천동매3(caspase3,CASP3).13개여세포증식화세포주기상관기인,분별시세포분렬주기단백27(cell division cycle27,CDC27)、CDC37、CDC28단백격매1B(CDC28 protein kinase 1B,CKS1B)、세포주기단백B1 (cycling B1,CCNB1)、CUL2(cullin 2)、CUL3(cullin 3)、CUL4A(cullin 4A)、생장억제화DNA손상유도기인45A (growth arrest and DNA damage-inducible 45A,Gadd45A)、감수분렬중조11동원체B(meiotic recombination 11 homolog B,MRE11 B)、차두광M1 (forkhead box M1,FOXM1)、미형염색체7(minichromosome maintenance 7,MCM7)、단극륭항체Ki-67항원(antigen identified by monoclonal antibody Ki67,MKI67)화세포증식핵항원(proliferating cell nuclear antigen,PCNA).11개여세포응격화독성반응유관적기인분별시곡광감태과양화물매1(glutathione peroxidase 1,GPX1)、금속류단백1A(metallothionein,MT1A)、초양화물기화매1(superoxide dismutase-1,SOD1)、열휴극단백A1A(heatshock protein A1A,HSPA1A)、HSPA2、HSPA4、HSPA5、HSP9B、HSPCA、거서세포유주억제인자(macrophage migration inhibitory factor,MIF)화섬용매원격활물억제인자1(plasminogen activator inhibitor,PAI1).결론 평양매소치료영유인혈관류시다인소、다수평종합작용적결과,평양매소능구작용우세포DNA적다개편단,기능유발세포조망,억제세포적증식,환능직접사세포적응격여독성반응능력강저,세포독성작용화유도세포조망시동일작용궤제적량충표현형식,병비량충작용궤제.
Objective To investigate the therapeutic mechanism of Bleomycin A 5 on infancy hemangioma-Methods After intralesional injection of Bleomycin A5 into the tumor of animal model of infancy hemangioma,the variation of tumor form was and the variation of tumor structure were observed using light microscope and electron microscope,the variation of tumor gene expression spectra was also tested by DNA microarray technique.Results After treatment,the tumor gradually shrunk,hardened,disappeared one month later.The tumor lost appearance of infancy hemangioma and replaced by lamellar collagen fibers and cellular nucleus scattered in the fibers,and almost all cells were necrotic and dissolved.Under electron microscope,only large stretches of dissolved cell could be seen without intact cells and blood vessels,but apoptotic cells and bodies could also be found.The results of DNA microarray analysis showed that 9 genes associated with apoptosis (murine double minute 2,heat-labile enterotoxin B subunit,lymphotoxin B receptor,tumor necrosis factor ligand superfamily 7,tumor necrosis factor receptor superfamily 21,tumor necrosis factor receptor superfamily 1 A,myeloid cell leukemia-1,caspase3),13 genes associated with cell proliferation and cell cycle(cell division cycle27,cell division cycle37,CDC28 protein kinase 1B,cycling B1,cullin 2,cullin 3,cullin 4A,growth arrest and DNA damage-inducible 45A,meiotic recombination 11homolog B,forkhead box Ml,minichromosome maintenance7,antigen identified by monoclonal antibody ki 67,proliferating cell nuclear antigen),and 11 genes associated with cellular stress and toxic reaction (glutathione peroxidase 1,metallothioneins,uperoxide dismutase-1,heat shock protein A1 A,heat shock protein A2,heat shock protein A4,heat shock protein A5,heat shock protein 9B,heat shock protein CA,macrophage migration inhibitory factor,plasminogen activator inhibitor)were up or down regulated more than 2 folds in tumors treated with Bleomcyin A5 compared with controls.Conclusions The therapeutic effect of Bleomycin A5 on infancy hemangioma is the synthetic results of multiple factors.Bleomycin A5 could not only induce apoptosis and inhibit cell proliferation,but also depressed the ability of cell stress and toxic reaction.
