Shukla Neha, Kumari Shivani, Verma Poornima, Srivastava Amisha, Singh Neelam, Rao Gyan Manjary, Prasad Narayan, Gupta Sushil, Srivastava Anshika, Ansari M S, Janakiraman Soorya, Baden Marey, Wolkenhauer Olaf, Gupta Shailendra K, Gautam Naveen Kumar
Department of Urology and Renal Transplantation, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India.
Department of Nephrology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India.
J Cell Mol Med. 2025 Sep;29(17):e70819. doi: 10.1111/jcmm.70819.
IL10 is a very effective anti-inflammatory cytokine. IL10 imbalance is linked to type 2 diabetes mellitus (T2DM) and also to renal hypertrophy, glomerular membrane thickening, and onset of diabetic nephropathy (DN). We aimed to investigate the association of IL10 gene polymorphism (rs1800871T/C, rs1800896A/G) with DN and determine the influence of variants on its expression level and interaction with transcription factors. We genotyped 301 study subjects, comprising 75 DN, 126 T2DM patients, and 100 controls. All were analysed for biochemical assays and genotypic analysis by PCR-RFLP and confirmed by Sanger sequencing. The haplotype analysis was calculated by Chi-square test. mRNA expression and its correlation with variants were assessed by using RT-PCR. Statistical analyses were done by using SPSS and GraphPad software. Screening of transcription factors with IL10 gene variants was performed using the TRANSFAC database, and the variants impact on IL10 molecular interactions was analysed. This study revealed that IL10 gene polymorphism rs1800896 was significantly associated with DN. 'CG' and 'TG' haplotypes were significantly associated with DN. Expression levels of IL10 were upregulated in DN patients. The genetic correlation study shows that IL10 gene expression was downregulated in the rs1800871 alternate variant genotype (CC) while upregulated in the rs1800896 alternate variant genotype (GG). In silico analyses suggest that the binding affinity of transcription factor CEBPA decreases due to the rs1800871 alternate variant, while the affinity of KLF4 increases in the case of the rs1800896 alternate variant. Our in silico results correlated with IL10 expression analysis in respective patient groups. Overall, our findings highlight the role of IL10 gene polymorphism in DN progression in the North Indian population.
白细胞介素10（IL10）是一种非常有效的抗炎细胞因子。IL10失衡与2型糖尿病（T2DM）相关，也与肾肥大、肾小球膜增厚以及糖尿病肾病（DN）的发病有关。我们旨在研究IL10基因多态性（rs1800871T/C、rs1800896A/G）与DN的关联，并确定这些变异对其表达水平以及与转录因子相互作用的影响。我们对301名研究对象进行了基因分型，其中包括75名DN患者、126名T2DM患者和100名对照。所有研究对象均通过聚合酶链反应-限制性片段长度多态性（PCR-RFLP）进行生化检测和基因分型分析，并通过桑格测序进行确认。单倍型分析通过卡方检验计算。使用逆转录-聚合酶链反应（RT-PCR）评估mRNA表达及其与变异的相关性。使用SPSS和GraphPad软件进行统计分析。使用TRANSFAC数据库筛选与IL10基因变异相关的转录因子，并分析这些变异对IL10分子相互作用的影响。本研究表明，IL10基因多态性rs1800896与DN显著相关。“CG”和“TG”单倍型与DN显著相关。DN患者中IL10的表达水平上调。遗传相关性研究表明，在rs1800871的替代变异基因型（CC）中IL10基因表达下调，而在rs1800896的替代变异基因型（GG）中表达上调。计算机模拟分析表明，由于rs1800871的替代变异，转录因子CEBPA的结合亲和力降低，而在rs1800896替代变异的情况下，KLF4的亲和力增加。我们的计算机模拟结果与各患者组中的IL10表达分析相关。总体而言，我们的研究结果突出了IL10基因多态性在北印度人群DN进展中的作用。
