摘要
目的通过检测E2F1基因在高氧致早产儿慢性肺疾病(CLD)新生鼠肺成纤维细胞中的动态表达,初步探讨E2F1基因在CLD肺间质纤维化发生发展过程中的生物学作用。方法足月新生SD大鼠于生后12 h内分别持续吸入85%高氧和空气,于3、7、14、21 d随机处死动物后,无菌条件下采集肺组织,进行肺成纤维细胞的原代培养,免疫组化、Western blot法检测E2F1蛋白表达r,eal-time PCR法检测E2F1 mRNA含量。结果生后3 d空气组和高氧组E2F1蛋白及mRNA的表达(蛋白:0.251±0.054,0.249±0.074;mRNA:0.066 4±0.003 4,0.066 8±0.003 7)无统计学差异(P>0.05);生后7 d时高氧组E2F1蛋白及mRNA的表达较空气组增加(蛋白:0.248±0.041,0.278±0.034;mRNA:0.065 9±0.003 1,0.072 8±0.004 7),差异有统计学意义(P<0.05);14 d、21 d时增加明显(蛋白:0.243±0.077 vs 0.328±0.057,0.241±0.038 vs 0.377±0.063;mRNA:0.241±0.038 vs 0.377±0.063,0.065 5±0.003 8 vs 0.075 0±0.004 1,P<0.01)。结论 E2F1异常高表达参与了高氧致CLD鼠肺成纤维细胞过度增殖的病理过程,在肺间质纤维化的发生发展过程中发挥重要作用。
Objective To determine the dynamic expression of E2F1 gene in lung fibrolasts of neonatal rats with chronic lung disease（CLD）,and to elucidate the biological effects of E2F1 in pulmonary interstitial fibrosis of CLD.Methods Full-term newborn SD rats were continuously exposed to 85% oxygen（hyperoxia group） or room air（room air group） within 12 hours after birth.The rat pups were randomly selected to be killed on postnatal day 3,7,14 and 21,lungs were immediately resected under sterile condition and primary culture of lung fibrolasts was performed.The expressions of E2F1 protein and mRNA in lung fibrolasts were determined by immunohistochemistry,Western blot and Real-time PCR,respectively.Results No significant difference in the expressions of E2F1 protein and mRNA was found between hyperoxia and room air groups on postnatal day 3（protein：0.251±0.054 vs 0.249±0.074;mRNA：0.066 4±0.003 4 vs 0.066 8±0.003 7,P 〉0.05）.Compared with room air groups,the expression of E2F1 in hyperoxia groups began to decrease since postnatal day 7（protein：0.248±0.041 vs 0.278±0.034;mRNA：0.065 9±0.003 1 vs0.072 8±0.004 7,P 〈 0.05） and significantly decreased on postnatal day 14 and 21（protein：0.243±0.077 vs 0.328±0.057,0.241±0.038 vs 0.377±0.063;mRNA：0.241±0.038 vs 0.377±0.063,0.065 5±0.003 8 vs 0.075 0±0.004 1,P 〈 0.01）.Conclusion Abnormality of E2F1 expression is involved in the pathological process of the proliferation of lung fibroblasts in hyperoxia-induced neonatal rats CLD and plays an important role in lung fibrosis.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2011年第2期109-112,共4页
Journal of China Medical University
基金
辽宁省教育厅高校科研计划项目(2010599)
