Namuleme Cissy B, Kato Charles D, Kasozi Dennis M
Department of Biotechnical and Laboratory Sciences, School of Biosecurity, College of Veterinary Medicine, Animal Resources and Biosecurity, Makerere University, P.O. Box 7062, Kampala, Uganda.
Department of Biochemistry and Systems Biology, School of Biosciences, College of Natural Sciences, Makerere University, P.O. Box 7062, Kampala, Uganda.
Biol Res. 2025 Sep 3;58(1):61. doi: 10.1186/s40659-025-00639-w.
Sickle cell disease (SCD) is characterised by chronic oxidative stress. However, there is limited information on how polymorphisms in cytokine genes influence oxidative stress in SCD patients. The study aimed to determine the effect of Interleukin gene (IL-10) and Tumor Necrosis Factor (TNF) polymorphisms on oxidative stress and cytokine levels in SCD patients from Mulago hospital.
A case control study with cross-sectional sample size of 163 SCD patients and 189 healthy controls was carried out. The extent of oxidative stress was quantified using Malondialdehyde (MDA) by spectrophotometry. Levels of IL-10 and TNF-α were measured using the Enzyme-Linked Immuno-Sorbent Assay (ELISA). The Amplification Refractory Mutation System polymerase chain reaction (ARMS-PCR) assay was used to genotype IL10-1082 A > G, (rs1800896), IL10-819 C > T (rs1800871) and TNF-α-308G > A (rs1800629) and TNF-β + 252 A > G (rs909253) gene polymorphisms.
Samples showed significantly (P = 0.0063) higher median plasma levels of MDA in SCD patients (2.756µM) than healthy controls (2.364µM). A similar trend was observed with significantly (P < 0.0001) higher median plasma levels of IL-10 in SCD patients (20.37pg/ml) than healthy controls (7.5pg/ml). The most frequent genotype for IL-10 (-1082, rs1800896) gene polymorphism was heterozygous GA (62.6%). No significant association between IL10 (-1082G > A, rs1800896) gene polymorphisms and SCD was observed (OR = 1.08, 95% CI = 0.54-2.14, P = 0.87). Yet, IL10 homozygous GG (-1082, rs1800896) (22.12pg/ml) that was found to be significantly associated (P = 0.0234) with increased plasma levels of IL-10 as compared to heterozygous genotype (GA) (13.94pg/ml) in SCD patients. Similarly, higher levels of MDA were found to be significantly (P < 0.0001) associated with homozygous GG at IL-10 (-1082, rs1800896). The most frequent and only reported genotype for TNF-α/β gene polymorphisms were heterozygous GA, thus no associations were described.
In conclusion, our results suggest that the IL10 (-1082 G > A, rs1800896) gene polymorphism is associated with increased oxidative stress and IL-10 cytokine level in Ugandan SCD patients.
镰状细胞病（SCD）的特征是慢性氧化应激。然而，关于细胞因子基因多态性如何影响SCD患者氧化应激的信息有限。本研究旨在确定白细胞介素基因（IL-10）和肿瘤坏死因子（TNF）多态性对穆拉戈医院SCD患者氧化应激和细胞因子水平的影响。
进行了一项病例对照研究，横断面样本量为163例SCD患者和189例健康对照。采用分光光度法通过丙二醛（MDA）对氧化应激程度进行定量。使用酶联免疫吸附测定（ELISA）测量IL-10和TNF-α水平。采用扩增阻滞突变系统聚合酶链反应（ARMS-PCR）分析对IL10 -1082 A＞G（rs1800896）、IL10 -819 C＞T（rs1800871）以及TNF-α -308G＞A（rs1800629）和TNF-β +252 A＞G（rs909253）基因多态性进行基因分型。
样本显示，SCD患者（2.756µM）血浆MDA中位数水平显著高于健康对照（2.364µM）（P = 0.0063）。观察到类似趋势，SCD患者（20.37pg/ml）血浆IL-10中位数水平显著高于健康对照（7.5pg/ml）（P＜0.0001）。IL-10（-1082，rs1800896）基因多态性最常见的基因型是杂合子GA（62.6%）。未观察到IL10（-1082G＞A，rs1800896）基因多态性与SCD之间存在显著关联（OR = 1.08，95%CI = 0.54 - 2.14，P = 0.87）。然而，在SCD患者中，发现IL10纯合子GG（-1082，rs1800896）（22.12pg/ml）与血浆IL-10水平升高显著相关（P = 0.0234），相比杂合子基因型（GA）（13.94pg/ml）。同样，发现较高水平的MDA与IL-10（-1082处，rs1800896）的纯合子GG显著相关（P＜0.0001）。TNF-α/β基因多态性最常见且唯一报道的基因型是杂合子GA，因此未描述其关联。
总之，我们的结果表明，IL10（-1082 G＞A，rs1800896）基因多态性与乌干达SCD患者氧化应激增加和IL-10细胞因子水平相关。
