Department of Neurology, The Third Affiliated Hospital of Qiqihar Medical University, Qiqihar, China.
Institute of Medical Sciences, Qiqihar Medical University, Qiqihar, China.
Minerva Med. 2021 Apr;112(2):261-268. doi: 10.23736/S0026-4806.20.06462-9. Epub 2020 Mar 20.
This study aimed to investigate the effect of cobalt transporter II gene (rs1801198, rs2301957, rs9606756) polymorphism on serum homocysteine level and its correlation with young and middle recurrent cerebral infarction.
A total of 348 young and middle-aged patients with cerebral infarction admitted to The Third Affiliated Hospital of Qiqihar Medical University from January 2017 to March 2018 were enrolled. The patients were divided into recurrent and non-recurrent groups according to follow-up. DNA was extracted from the peripheral blood of patients, and the DNA samples were genotyped by IlluminaBeadArray technology to detect the gene polymorphisms of cobalt transporter II (TCN2) sites (rs1801198, rs2301957, rs9606756), and the homocysteine (hcy) level was determined by cyclic enzymatic method. VitB12 and folate levels were measured by chemiluminescence immunoassay, and holo transcobalamin (holoTC) expression levels were detected by enzyme-linked immunosorbent assay.
The frequency of alleles of rs9606756 mutation in the recurrent group was higher than that in the non-recurrent group (P<0.05), and the Hcy level in rs9606756 locus genotype AG+GG was significantly higher than the AA genotype in the recurrent group (P=0.031). Pearson correlation analysis showed that Hcy levels were associated with different genotypes of rs9606756 in the recurrent group (r=0.483, P=0.0003). The rs9606756 allele AA in SH-SY5Y cells was replaced with GG by point mutation experiment. The Hcy metabolism levels of wild and mutant cells were detected. The accumulation level of Hcy in the mutant group was significantly increased (P=0.007). The holoTC in the supernatant was significantly reduced in the mutant (P=0.032).
The TCN2 gene rs9606756 mutation is closely related to the level of Hcy metabolism in young and middle-aged patients, which may affect the recurrence of cerebral infarction. It is of great significance to further understand the pathogenesis, prevention and treatment of recurrent cerebral infarction in young and middle-aged patients.
本研究旨在探讨钴转运蛋白 II 基因（rs1801198、rs2301957、rs9606756）多态性对血清同型半胱氨酸水平的影响及其与中青年复发性脑梗死的相关性。
选取 2017 年 1 月至 2018 年 3 月齐齐哈尔医学院附属第三医院收治的中青年脑梗死患者 348 例，根据随访结果分为复发组和非复发组。抽取患者外周血提取 DNA，采用 IlluminaBeadArray 技术对钴转运蛋白 II（TCN2）位点（rs1801198、rs2301957、rs9606756）的基因多态性进行检测，采用循环酶法测定同型半胱氨酸（hcy）水平，采用化学发光免疫法测定维生素 B12 和叶酸水平，采用酶联免疫吸附法检测全同型半胱氨酸（holoTC）表达水平。
复发组 rs9606756 突变等位基因频率高于非复发组（P<0.05），且 rs9606756 位点基因型 AG+GG 的 Hcy 水平明显高于复发组的 AA 基因型（P=0.031）。Pearson 相关分析显示，复发组 Hcy 水平与 rs9606756 不同基因型相关（r=0.483，P=0.0003）。通过点突变实验将 SH-SY5Y 细胞 rs9606756 等位基因 AA 替换为 GG。检测野生型和突变型细胞的 Hcy 代谢水平，突变组 Hcy 积累水平明显升高（P=0.007），突变组上清液中的 holoTC 明显减少（P=0.032）。
TCN2 基因 rs9606756 突变与中青年患者 Hcy 代谢水平密切相关，可能影响脑梗死的复发，对进一步了解中青年患者复发性脑梗死的发病机制、预防和治疗具有重要意义。
