Nishi Hiroshi, Inagi Reiko, Kato Hideki, Tanemoto Masayuki, Kojima Ichiro, Son Daisuke, Fujita Toshiro, Nangaku Masaomi
Division of Nephrology and Endocrinology, University of Tokyo School of Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan.
J Am Soc Nephrol. 2008 Aug;19(8):1500-8. doi: 10.1681/ASN.2007101085. Epub 2008 Apr 30.
Hemoglobin (Hb) serves as the main oxygen transporter in erythrocytes, but it is also expressed in nonhematopoietic organs, where it serves an unknown function. In this study, microarray and proteomic analyses demonstrated Hb expression in the kidney. Rat kidneys were perfused extensively with saline, and glomeruli were isolated by several techniques (sieving, manual dissection, and laser capture-microdissection). Reverse transcriptase-PCR revealed glomerular alpha- and beta-globin expression, and immunoblotting demonstrated expression of the protein. In situ hybridization studies showed expression of the globin subunits in the mesangium, and immunostaining confirmed this localization of Hb. Furthermore, globin mRNA expression was detected in primary cultures of rat mesangial cells but not in cultured glomerular endothelial or epithelial cells. For investigation of Hb function in mesangial cells, the SV40-MES13 murine mesangial cell line was transfected with a vector expressing alpha- and beta-globins; this overexpression reduced production of hydrogen peroxide-induced intracellular radical oxygen species and enhanced cell viability against oxidative stress. In summary, Hb is expressed by rat mesangial cells, and its potential functions may include antioxidative defense.
血红蛋白(Hb)是红细胞中的主要氧气转运蛋白,但它也在非造血器官中表达,其功能尚不清楚。在本研究中,微阵列和蛋白质组学分析表明肾脏中存在Hb表达。用生理盐水对大鼠肾脏进行广泛灌注,然后通过多种技术(筛分、手工解剖和激光捕获显微切割)分离肾小球。逆转录聚合酶链反应(RT-PCR)显示肾小球中有α-和β-珠蛋白表达,免疫印迹法证实了该蛋白的表达。原位杂交研究表明系膜中有珠蛋白亚基表达,免疫染色证实了Hb的这种定位。此外,在大鼠系膜细胞的原代培养物中检测到珠蛋白mRNA表达,但在培养的肾小球内皮细胞或上皮细胞中未检测到。为了研究Hb在系膜细胞中的功能,用表达α-和β-珠蛋白的载体转染SV40-MES13小鼠系膜细胞系;这种过表达减少了过氧化氢诱导的细胞内活性氧的产生,并增强了细胞对氧化应激的生存能力。总之,大鼠系膜细胞表达Hb,其潜在功能可能包括抗氧化防御。