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目的:探讨热休克蛋白70-1(HSP70-1)基因多态性对中暑易感性的影响。方法:采用病例-对照1∶2研究设计。采用PCR产物直接测序法检测HSP70-1基因rs1043618(+190G/C)多态性的基因型。包含不同等位(G/C)的基因片段插入pGL3-Basic以构建等位特异性重组表达载体,并分别与内对照质粒pRL-TK共转染人脐静脉内皮细胞株,应用荧光素酶报告基因分析法比较不同等位基因重组载体在热应激情况下的相对荧光素酶表达活性。结果:rs1043618(+190G/C)位点GG、GC和CC基因型表达与对照组比较,CC基因型在病例组中的频率明显升高,差异有统计学意义(11.5%vs.4.4%,P=0.006)。在显性模式下,携带C等位基因的患者在病例组中的频率明显高于对照组(53.1%vs.40.7%,P=0.031)。荧光素酶报告实验显示,与+190C等位基因重组载体相比,在热应激状态下+190G等位基因重组载体可明显增强荧光素酶活性(P=0.012)。结论:HSP70-1基因rs1043618多态性与中暑易感性有关,其原因可能在于影响了HSP的表达水平。
Objective: To investigate the effect of heat shock protein 70-1 (HSP70-1) gene polymorphism on susceptibility to heat stroke. Methods: Case-control 1: 2 study design. The genotypes of HSP70-1 gene rs1043618 (+ 190G / C) polymorphism were detected by PCR direct sequencing. The gene fragment containing different alleles (G / C) was inserted into pGL3-Basic to construct an allelic-specific recombinant expression vector and cotransfected into human umbilical vein endothelial cell line with the internal control plasmid pRL-TK separately. Luciferase reporter Gene analysis was used to compare the relative luciferase expression activities of different allelic recombination vectors under heat stress. Results: The frequencies of genotypes GG, GC and CC in rs1043618 (+190 G / C) group were significantly higher than those in control group (P <0.05). The frequency of CC genotype was significantly higher in case group (11.5% vs.4.4% P = 0.006). In dominant mode, the frequency of patients carrying the C allele was significantly higher in the case group than in the control group (53.1% vs. 40.7%, P = 0.031). The luciferase reporter assay showed that the + 190G allele recombinant vector significantly enhanced luciferase activity (P = 0.012) under heat stress compared with the + 190C allele recombinant vector. Conclusion: The rs1043618 polymorphism of HSP70-1 gene is associated with the susceptibility to heat stroke, which may be due to the influence of HSP expression.