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目的:检测大鼠胚胎心脏发育过程中凋亡及凋亡基因表达,进一步明确心脏发育的调控机制。方法:采用半定量RT-PCR、免疫组织化染色SABC法和TUNEL法对E11～E19 SD大鼠心脏凋亡细胞和p53mRNA、Bax蛋白表达进行测定。结果:p53和BAX在E11～E19表达均呈逐渐增强再减弱的趋势,表达的高峰位于E14。TUNEL染色结果显示,E11～E19均可见凋亡小体。E11时散在分布于心内外膜;E13主要分布于心尖;E14时表达最强,主要分布于心房和心腔分隔处;E16后表达减弱,分布主要集中于心腔内壁。结论:大鼠心脏发育中后期均有凋亡存在;E14为凋亡表达高峰时期;p53和Bax在促进心肌细胞凋亡、心脏外形和心腔形成方面起到重要的作用。
OBJECTIVE: To detect the expression of apoptosis and apoptosis genes in rat embryonic heart during development, and to further clarify the regulatory mechanism of cardiac development. Methods: The expression of p53 mRNA and protein of Bax in the heart of E11-E19 SD rats were determined by semi-quantitative RT-PCR, immunohistochemical SABC method and TUNEL method. Results: The expression of p53 and BAX in E11 ~ E19 showed a trend of increasing and then decreasing. The peak of expression was located at E14. TUNEL staining showed that apoptotic bodies were seen in E11 ~ E19. E11 scattered in the heart and adventitia; E13 mainly distributed in the apex; E14 strongest expression, mainly distributed in the atrium and heart chamber; E16 expression decreased, the distribution is mainly concentrated in the heart chamber. Conclusion: There is apoptosis in the middle and late stages of cardiac development in rats. E14 is the peak period of apoptosis expression. P53 and Bax play an important role in promoting cardiomyocyte apoptosis, cardiac appearance and cardiac cavity formation.