骨形成蛋白-4参与调控胚胎舌形态发育  

Bone morphogenetic protein-4 regulates embryonic tongue morphogenesis

作　　者：周彦秋[1] 林久祥[1] 

机构地区：[1]北京大学口腔医学院.口腔医院正畸科,北京100081

出　　处：《北京大学学报（医学版）》2009年第1期76-79,共4页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(30572067)资助~~

摘　　要：目的:采用胚胎舌体外培养结合扫描电子显微镜、免疫组织化学方法,探讨骨形成蛋白(bone morphoge-netic protein-4,BMP4)对舌体形态发育的影响。方法:分别向标准培养基中添加不同浓度的BMP4及其拮抗剂Noggin,以所加因子及其浓度的不同分为6个实验组,BMP4分为3组,浓度分别为0.03 mg/L,0.3 mg/L,1 mg/L;BMP4的拮抗剂Noggin分为3组,浓度分别为1 mg/L,3 mg/L,10 mg/L;标准培养基培养组为对照组。将解剖获得的大鼠胚胎第13天(embryonic day 13,E13)的胚胎舌在培养基中格栅法培养3 d,固定后,用扫描电子显微镜观察记录胚胎舌;分别测量胚胎舌全长、舌前1/8、舌前1/4和舌体1/2宽度,统计分析测量结果。胚胎舌内放置经PBS和BMP4(667 mg/L)溶液处理的凝胶颗粒,以PBS组为对照组,在标准培养基中培养3 d后,制成冰冻切片,检测Ki67的表达。结果:(1)舌形态:舌体长度明显改变(P<0.05),相对对照组舌体长度(1 037.8±126.2μm),BMP4各浓度组舌体长度明显变短(0.03 mg/L组877.3±67.6μm,0.3 mg/L组838.5±88.9μm,1 mg/L组718.7±38.6μm),Noggin各浓度组变化不明显(1 mg/L组1 191.1±75.7μm,3 mg/L组1 169.8±108.7μm,10mg/L组1 241.3±17.4μm);舌前1/8宽度明显改变(P<0.05),相对对照组(639.1±106.2μm),BMP4各浓度组舌前1/8宽度明显变窄(0.03 mg/L组332.1±80.9μm,0.3 mg/L组305.1±51.3μm,1 mg/L组276.9±45.9μm),Noggin各浓度组除10 mg/L组外,舌前1/8宽度加宽(1 mg/L组815.5±90.3μm,3 mg/L组857.6±87.1μm,10 mg/L组807.1±113.8μm);舌前1/4宽度改变明显(P<0.05),相对对照组(653.7±101.6μm),BMP4各浓度组舌前1/4宽度明显变窄(0.03 mg/L组421.3±43.8μm,0.3 mg/L组407.3±15.6μm,1 mg/L组363.7±24.7μm),而Noggin各浓度组明显加宽(1 mg/L组838.0±130.5μm,3 mg/L组947.2±34.9μm,10mg/L组889.4±74.6μm);舌体1/2宽度变化明显(P<0.05),相对对照组(683.1±79.8μm),BMP4各浓度组舌体1/2宽度明显变窄(0.03 mg/L组567.3±35.8μm,0.3 mg/L组548.4±30.5μm,1 mg/L组457.4±48.0μObjective ： To analyze the effect of Bone morphogenesis 4 and its antagonist Noggin on morphogenesis of tongue. Methods： Dissected rats to get embryonic day 13 （E13） tongues; fed El3 tongues in standard medium, BMP4 （0. 03 mg/L, 0. 3 mg/L, 1 mg/L）,and the antgonist Noggin（1 mg/L, 3 mg/L, 10 rag/L） medium; cultured for 3 days; fixed samples, observed tongues with scanning electronic microscope （SEM） ; measured the whole tongue length , anterior 1/8,1/4 width and middle width of cultured tongues and analyzed data with SPSS 10. 0. To further study the effects of BMP4 on epithelial and mesenchymal cell proliferation, Affi-gel blue gel beads were applied. Beads were soaked in PBS and BMP4 （667 mg/L）, and implanted in the E13 embryonic tongues; then after cultured in standard medium for 3 days, tongues were embedded in O. C. T. and cut into 12 p.m series sections. Ki67 was detected by immunohistoehemical method. Results： （ 1 ） Whole length of tongues changed greatly （ P 〈 0.05 ）, the length was shortened in BMP4 groups （0. 03 mg/L group 877.3 ± 67.6 um , 0. 3 mg/L group 838.5 ±88.9 um,1 mg/L group 718.7 ±38.6 um） compared with standard medium （ 1037. 8 ± 126.2 um）, Noggin groups had no obvious change; the anterior 1/8 width of tongues changed siguificantly（P 〈 0. 05 ）, the anterior 1/8 width was narrower in BMP4 groups （0. 03 mg/L group 332. 1 ± 80. 9 um,0. 3 mg/L group 305.1 ±51.3 um,1 mg/L group 276.9 ±45.9 um） compared with standard group （639. 1 ±106. 2um）, except 10 mg/L group, Noggin groups were wider （ 1 mg/L group 815.5 ± 90. 3 um,3 mg/L group 857.6 ±87. 1um,10 mg/L group 807. 1 ±113.8 um） ; the anterior 1/4 width of tongue changed magnificantly, also （ P 〈 0. 05 ） , BMP4 groups were narrower （ 0. 03 mg/L group 421.3 ±43.8 um,0.3 mg/L group 407.3 ± 15.6 um,1 mg/L group 363.7 ±24. 7um） compared with standard group （ 653.7 ± 101.6 um） , whereas, Noggin groups were wider greatly （ 1 mg/L group 838.0 ± 130. 5 um,3 mg/L gro

关 键 词：骨形态发生蛋白质类 胚胎发育 舌 细胞增殖 

分 类 号：R322.41[医药卫生—人体解剖和组织胚胎学]