Kaplan Michael, Renbaum Paul, Hammerman Cathy, Vreman Hendrik J, Wong Ronald J, Stevenson David K
Department of Neonatology, Shaare Zedek Medical Center, Jerusalem, Israel.
Neonatology. 2014;106(4):323-9. doi: 10.1159/000365744. Epub 2014 Oct 1.
Heme oxygenase (HO) is the initial, rate-limiting enzyme in the conversion of heme to bilirubin. Dinucleotide (GT)n repeat length in the promoter region of the encoding gene modulates transcription: shorter alleles, in contrast with longer allele counterparts, are associated with greater gene expression and should result in increased heme catabolism.
We compared the rates of heme catabolism and plasma total bilirubin (TB) between HO-1 promoter genotypes of varying (GT)n repeat lengths in glucose-6-phosphate dehydrogenase (G6PD)-normal and -deficient neonates.
HO-1 promoter length was determined from genomic DNA from previous studies by size discrimination of fluorescently-labeled PCR products with capillary electrophoresis. Sizing was confirmed by sequencing homozygote samples. Alleles were categorized as: short (≤24 GT repeats), medium (25-33 GT repeats), and long (≥34 GT repeats). Previously determined values for blood carboxyhemoglobin, corrected for inspired carbon monoxide (COHbc), and TB were used to determine the rate of heme catabolism and 3rd day TB values for each HO-1 promoter length genotype, respectively. G6PD Mediterranean was determined by PCR analysis.
Neither COHbc nor TB values were significantly different between various HO-1 promoter genotypes for either G6PD-normal or -deficient neonates.
In the steady state, HO-1 promoter genotypes, based on the length of (GT)n repeats, do not modulate heme catabolism or 3rd day TB values in either G6PD-normal or -deficient neonates.
血红素加氧酶(HO)是血红素转化为胆红素过程中的首个限速酶。编码基因启动子区域的二核苷酸(GT)n重复长度可调节转录:与较长等位基因相比,较短等位基因与更高的基因表达相关,应会导致血红素分解代谢增加。
我们比较了葡萄糖-6-磷酸脱氢酶(G6PD)正常和缺乏的新生儿中不同(GT)n重复长度的HO-1启动子基因型之间的血红素分解代谢率和血浆总胆红素(TB)水平。
通过毛细管电泳对荧光标记的PCR产物进行大小区分,从先前研究的基因组DNA中确定HO-1启动子长度。通过对纯合子样本进行测序来确认大小。等位基因分为:短(≤24个GT重复)、中(25-33个GT重复)和长(≥34个GT重复)。先前测定的经吸入一氧化碳校正的血液碳氧血红蛋白(COHbc)值和TB值,分别用于确定每种HO-1启动子长度基因型的血红素分解代谢率和第3天的TB值。通过PCR分析确定G6PD地中海型。
对于G6PD正常或缺乏的新生儿,不同HO-1启动子基因型之间的COHbc值和TB值均无显著差异。
在稳态下,基于(GT)n重复长度的HO-1启动子基因型在G6PD正常或缺乏的新生儿中均不调节血红素分解代谢或第3天的TB值。