摘要
目的建立肝素结合性表皮生长因子(HB-EGF)的转基因动物模型,利用转基因动物模型研究HB-EGF在心脏功能中的作用。方法构建α-MHC-HB-EGF表达载体,将目的片段注射到受精卵的雄原核中,使其发育成携带有HB-EGF的转基因小鼠。通过PCR的方法鉴定转基因的首建鼠。采用Western blotting方法鉴定HB-EGF在心脏中的表达,取HB-EGF在心脏中特异性高表达的阳性转基因小鼠的F1代与同窝阴性对照小鼠的心脏做BrdU免疫组化和Masson染色。结果得到了两只HB-EGF阳性的首建鼠,Western blotting发现16号小鼠中HB-EGF在心脏中的表达与同窝阴性对照小鼠相比蛋白表达明显增加。转基因小鼠心脏BrdU标记的阳性细胞数明显多于阴性对照组的阳性细胞数,Masson染色胶原纤维明显少于同窝阴性对照的小鼠。结论HB-EGF的高表达可以促进心肌细胞的增殖,减少胶原纤维生成,抑制心肌的纤维化。
Objective To prepare heparin-binding epidermal growth factor-like growth factor （HB-EGF） transgenic mice and investigate the role of HB-EGF in the heart function in transgenie mice. Methods Transgenie mice were created by mieroinjeetion of α-MHC-HB-EGF construct into male pronucleus of the zygotes. The genomie phonetype of transgenie founders were identified by PCR. The expression of HB-EGF was analyzed by Western blotting. BrdU immunohistochemistry and Masson triehrome staining were used to analyze the differences between transgenie mice and nontransgenie littermate controls. Results Two founders of HB-EGF transgenic mice were established. One of HB-EGF transgenic independent lines exhibited high-level cardiacspecific transgene expression compared with that of nontransgenie littermate controls assessed by Western blotting. More BrdUlabeled cells were seen in the heart tissue of transgenie mice than in the wild type mice and a marked decrease of collagen was found between myoeytes in the transgenio than wild type mice hearts. Conclusion These results support that HB-EGF may promote eardiomyoeyte proliferation and inhibit fibroplasia in the myoeardium.
出处
《中国比较医学杂志》
CAS
2007年第4期187-191,I0001,共6页
Chinese Journal of Comparative Medicine
