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建立亚急性和慢性心衰SD大鼠模型,用RNA-RNA原位杂交方法观察新基因FWA116在心脏组织的表达。以重组质粒FWA116/PGEM-T线性cDNA为模板,在T7耵或SP6RNA聚合酶作用下,体外合成地高辛(DIG)半抗原标记的FWA116cRNA探针。结果表明:与对照组相比,该基因在两种心衰大鼠心脏的中、小动脉内皮组织的mRNA表达均明显增高;与Northern blot及免疫组化的结果相一致,这提示DIG标记cRNA原位杂交方法的敏感和可靠。同时FWA116基因在心衰缺血缺氧心脏动脉内皮中的转录和翻译水平的高表达,说明该基因可能在心衰的病理过程中起到重要的作用。
To establish sub-acute and chronic heart failure SD rat model, observe the expression of FWA116 in heart tissue by RNA-RNA in situ hybridization. The FWA116cRNA probe labeled with digoxigenin (DIG) hapten was synthesized in vitro using the recombinant plasmid FWA116 / PGEM-T cDNA as a template under T7 耵 or SP6 RNA polymerase. The results showed that: Compared with the control group, the mRNA expression of the gene in the middle and small arterial endothelium in both heart failure rats was significantly increased; consistent with the result of Northern blot and immunohistochemistry, which indicated that the DIG-labeled cRNA In situ hybridization method is sensitive and reliable. At the same time, the high expression of FWA116 gene in the transcription and translation level of heart artery in heart failure and hypoxia induced cardiac dysfunction, indicating that the gene may play an important role in the pathological process of heart failure.