摘要
目的探讨胰岛素样生长因子Ⅰ(IGFⅠ)影响体外培养的滋养细胞黏附作用的机理。方法从6例早孕患者人工流产的绒毛组织中获取滋养细胞,建立体外滋养细胞黏附模型,在无血清培养液中,加入浓度为10nmol/L的IGFⅠ进行培养处理。单纯加入IGFⅠ的细胞为IGFⅠ组;加入IGFⅠ及抗整合素αvβ3抗体(抗整合素抗体)的细胞为IGFⅠ+抗整合素组;不加IGFⅠ的细胞为对照组。采用酶联免疫检测仪测定吸光度(A)值,表示黏附细胞的相对数量。于扫描电镜下观察IGFⅠ组和对照组的细胞形态的变化,并采用免疫组化方法,检测该两组细胞中磷酸化黏着斑激酶的表达。结果IGFⅠ组的平均A值为0.491±0.049,明显高于对照组的0.198±0.022,两组比较,差异有统计学意义(P<0.01)。IGFⅠ+抗整合素组的平均A值为0.184±0.031,明显低于IGFⅠ组,两组比较,差异有统计学意义(P<0.01);但与对照组比较,差异无统计学意义(P>0.05)。扫描电镜观察显示,滋养细胞加入IGFⅠ后,与纤维粘连蛋白黏附局部,可见滋养细胞表面片状伪足形成,与对照组比较,片状伪足明显增多并伸展。免疫组化染色结果,滋养细胞加入IGFⅠ后,细胞膜及细胞质中磷酸化黏着斑激酶呈阳性表达。结论浓度为10nmol/L的IGFⅠ通过激活磷酸化黏着斑激酶,刺激滋养细胞表面片状伪足的形成并伸展,促进滋养细胞黏附到纤维粘连蛋白上,而抗整合素抗体则可明显抑制IGFⅠ的促黏附作用。
Objective To investigate the mechanism of insulin-like growth factor-I(IGF-Ⅰ) affecting adhesion of trophoblast cells in vitro. Methods Trophoblast cells were obtained from early gestation at artificial abortion to set up the in vitro trophoblast cell adhesion model. The trophoblast cells were incubated with or without 10 nmol/L IGF-Ⅰ and were divided into three groups (10 nmol/L IGF-Ⅰ, 10 nmol/L IGF-Ⅰ+αvβ3Ab, and control). The amount of adhered cells was assessed by examining absorbency using enzyme-linked immunoassay. Morphological changes were studied using scanning electron microscopy. The expression of phosphorylated focal adhesion kinase was determined by immunocytochemistry. Results After serum-starved trophoblast cells were incubated only with IGF-Ⅰ, the mean absorbency was 0.491±0.049, obviously higher than control 0.198±0.022 and the difference was dramatic (P<0.01). When cells were pre-treated with antibody against αvβ3 integrin and then incubated with IGF-Ⅰ, the mean absorbency was only 0.184±0.031, distinctly lower than that incubated with IGF-Ⅰ, and the difference was significant (P<0.01), however, compared with control, there was no significant difference (P>0.05). Scanning electron microscopy highlighted a dramatic increase in lamellipodial formation and extension in the IGF-Ⅰ treated cells compared with control. Immunocytochemistry staining showed phosphorylated focal adhesion kinase was expressed in the trophoblast cells treated with IGF-Ⅰ. Conclusions 10 nmol/L IGF-Ⅰ can significantly stimulate trophoblast cells adhesion to fibronectin,but antibody against αvβ3 integrin obviously blocks its adhesion. IGF-Ⅰ can stimulate lamellipodial formation and extension at the adhesion sites, and promote adhesion of trophoblast cells to fibronectin by activating phosphorylated focal adhesion kinase.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2005年第6期392-395,共4页
Chinese Journal of Obstetrics and Gynecology
基金
湖北省卫生厅基金资助项目(NX200404)
