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400042 重庆,陆军特色医学中心战伤休克与输血研究室
[ 刘良明,E-mail: liangmingliu@yahoo.com LIU Liangming, E-mail: liangmingliu@yahoo.com
收稿:2021-09-13,
修回:2021-11-02,
纸质出版:2022-03-15
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臧家涛, 李清晖, 吴跃, 等. 血红蛋白α、β亚基在大鼠不同类型血管组织中的表达[J]. 陆军军医大学学报, 2022,44(5):399-405.
Jiatao ZANG, Qinghui LI, Yue WU, et al. Expression of hemoglobin α and β subunits in different types of rat vasculatures[J]. Journal of Army Medical University, 2022, 44(5): 399-405.
臧家涛, 李清晖, 吴跃, 等. 血红蛋白α、β亚基在大鼠不同类型血管组织中的表达[J]. 陆军军医大学学报, 2022,44(5):399-405. DOI: 10.16016/j.2097-0927.202109095.
Jiatao ZANG, Qinghui LI, Yue WU, et al. Expression of hemoglobin α and β subunits in different types of rat vasculatures[J]. Journal of Army Medical University, 2022, 44(5): 399-405. DOI: 10.16016/j.2097-0927.202109095.
目的
2
阐明大鼠血管组织是否表达血红蛋白(hemoglobin,Hb)α、β亚基,以及不同类型血管组织中的表达是否具有规律性。
方法
2
通过RT-PCR、DNA测序和Western blot分析SD大鼠颈总动脉、颈外静脉、胸主动脉、肠系膜上动脉、肠系膜上静脉、股动脉、股静脉等血管组织是否表达Hb α、β亚基,采用探针qPCR分析其表达差异,激光共聚焦荧光显微技术分析表达Hb α、β亚基的细胞类型。
结果
2
从上述血管组织中都扩增出与GenBank收录的大鼠Hb α、β亚基mRNA序列一致的DNA片段,同时通过Western blot检测到特异的Hb α、β亚基蛋白条带。qPCR结果显示不同血管组织中Hb α、β亚基的相对表达量存在差异,均在股静脉中表达最高(Hb α=7.23,Hb β=4.73),胸主动脉表达最低(Hb α=0.12,Hb β=0.11),且与其他血管比较差异有统计学意义(
P
<
0.01)。免疫荧光分析显示Hb α、β亚基由血管内皮细胞表达。
结论
2
Hb α、β亚基共同在大鼠传导、阻力、容量等不同类型血管组织中表达,其规律为相对表达量在阻力、容量血管中较高,而在传导血管中较低。
Objective
2
To clarify whether hemoglobin (Hb) α and β subunits are expressed in rat blood vessels
and whether there are differences in expression levels between different types of vasculatures.
Methods
2
RT-PCR
DNA sequencing and Western blotting were used to detect the expression of Hb α and β subunits at mRNA and protein levels in common carotid artery
thoracic aorta
superior mesenteric artery
femoral artery
external jugular vein
superior mesenteric vein and femoral vein in SD rats. Relative expression levels of Hb α and β subunits in these vessels were determined by probe qPCR. The results of immunofluorescence assay for Hb α and β subunits in these vessels was further observed by laser confocal microscopy.
Results
2
Specific DNA fragments were amplified from different blood vessels by RT-PCR
and the sequences of these DNA fragments were identical to the mRNA sequences of rattus norvegicus Hb α and β subunits in GenBank database respectively. Protein peptides with correct molecular weights were detected using antibodies against Hb α and β subunits respectively from total protein extracts of these blood vessels by Western blotting. qPCR results indicated that the relative expression levels of Hb α and β subunits varied between different blood vessels
and it was significantly lower in the thoracic aorta (Hb α=0.12
Hb β=0.11) while higher in the femoral vein (Hb α=7.23
Hb β=4.73) (
P
<
0.01). The results of immunofluorescence laser confocal microscopy indicated that Hb α and β subunits were expressed in vascular endothelial cells.
Conclusion
2
Hemoglobin α and β subunits are co-expressed in rat vasculatures
including conduit
resistance and capacitance vessels
and the relative expression level is higher in the resistance and capacitance vessels but lower in the conduit vessels.
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