Toll样受体4基因多态性与2型糖尿病并发肺结核患者发生糖尿病性下肢血管病变的关系研究

Association of ToII-like receptor 4 gene polymorphism with diabetic lower extremity vascular disease in patients with type 2 diabetes mellitus complicated with pulmonary tuberculosis

目的探索Toll样受体4（Toll-like receptor 4，TLR4）基因多态性与2型糖尿病并发肺结核（type 2 diabetes mellitus complicated with pulmonary tuberculosis，T2DM-PTB）患者发生糖尿病性下肢血管病变（lower extremity arterial disease，LEAD）的关系。方法选取2013年9月至2015年6月期间，在黑龙江省传染病防治院就诊的T2DM-PTB发生LEAD的患者87例（观察组）及T2DM-PTB未发生LEAD患者68例（对照组），所有患者于纳入研究时采集基本信息及血液样本。基因组DNA提取试剂盒提取血液DNA，采用实时荧光定量PCR技术（Taqman探针法）对TLR4基因的3个基因位点（rs7873784、rs1927911及rs1927914）进行基因多态性检测。结果观察组和对照组的TLR4基因单核苷酸多态性（single nucleotide polymorphisms，SNPs）位点等位基因分布：rs7873784位点G等位基因两组分别为91．38％（159／174）、91．18％（124／136），C等位基因两组分别为8．62％（15／174）、8．82％（12／136），两组间差异无统计学意义（χ2=0．00，P=0．950）；rs1927911位点T等位基因两组分别为59．20％（103／174）、61．76％（84／136），C等位基因两组分别为40．80％（71／174）、38．24％（52／136），两组间差异无统计学意义（χ2=0．21，P=0．646）；rs1927914位点T等位基因两组分别为59．77%（104／174）、64．71％（88／136），C等位基因两组分别为40．23％（70／174）、35．29％（48／136），两组间差异无统计学意义（χ2=0．79，P=0．374）。观察组和对照组的TLR4基因SNP位点基因型分布：rs7873784位点GG基因型两组分别为82．76％（72／87）、82．35％（56／68），GC基因型两组分别为17．24％（15／87）、17．65％（12／68），两组间差异无统计学意义（χ2=0．00，P=0．947）；rs1927911位点TT基因型两组分别为32．18％（28／87）、35．30％（24／68），CT基因型两组分别为54．02％（47／87）、52．94％（36／68），CC基因型两组分别为13．80％（12／87）、11．76％（8／68），两组间差异无统计学意义（χ2=0．24，P=0．887）；rs1927914位点TT基因型两组分别为33．33％（29／87）、42．65％（29／68），CT基因型两组分别为52．87％（46／87）、44．12％（30／68），CC基因型两组分别为13．80％（12／87）、13．23％（9／68），两组间差异无统计学意义（χ2=1．49，P=0．475）。TLR4基因rs7873784位点：与野生纯合基因型GG相比，GC基因型的调整OR值为0．90（95％CI：0．39～2．06，P=0．80）。rs1927911位点：与野生纯合基因型TT相比，CT基因型及CC基因型的调整OR值分别为1．00（95％CI：0．51～1．97，P=1．00）和1．29（95％CI：0．46～3．66，P=0．63）；rs1927914位点：与野生纯合基因型TT相比，CT基因型及CC基因型的调整OR值分别为1．27（95％CI：0．64～2．51，P=0．49）和1．24（95％CI：0．47～3．27，P=0．67）。3个基因位点各基因型间的差异均无统计学意义（P值均〉0．05）。结论TLR4基因rs7873784、rs1927911和rs1927914位点多态性与T2DMTB患者发生LEAD无关。

Objective To investigate the relationship between TLR4 gene polymorphism and LEAD in patients with T2DM-PTB. Methods We recruited 87 cases with T2DM-PTB patients complicated with LEAD and 68 cases with T2DM-PTB from Infectious Hospital of Heilongjiang Province between September 2013 and June 2015. The basic information of both T2DM-PTB patients complicated with LEAD and T2DM-PTB patients were collected. Meanwhile, peripheral blood was collected in order to undergo gene polymorphism analysis. DNA was extracted by genomic DNA extraction kits and genotyping was accomplished by using Real-time PCR （Taqman probe method）. Three TLR4 gene loci （rs 7873784, rs1927911, rs1927914） were tested. Results Allele distribution of TLR4 gene SNPs locus in the observation group and the control group： The G allele of rs7873784 were 91.38% （159/174） and 91.18% （124/136） in the two groups, and the C allele were 8.62% （15/174） and 8.82% （12/136） in the two groups, respectively. There was no significant difference between the two groups （χ2= 0.00, P= 0. 950）; The T allele of rs1927911 in two groups were 59.20% （103/174） and 61.76% （84/136）, respectively, and the C allele were 40. 80% （71/174） and 38.24G （52/136）, respectively. There was no significant difference between the two groups （χ2 =0.21,P=0. 646） The T allele of rs1927914 in two groups were 59.77% （104/174） and 64.71% （88/136）, respectively, and the C allele were 40. 23% （70/174） and 35.29% （48/136）, respectively. There was no significant difference between the two groups （χ2 =0.79,P=0. 374）. Genotype distribution of TLR4 gene SNP locus in the observation group and the control group： The GG genotypes of rs7873784 were 82.76% （72/87） and 82.35% （56/68） in two group, and the GC genotype of the two groups were 17.24% （15/87） and 17.65% （12/68）, respectively. The difference between the two groups was not statistically significant （χ2= 0. 00, P= 0. 947）. The TT genotypes of rs1927911 were 32.18% （28/87） and 35. 30% （24/68） in two groups, the CT genotype of the two groups were 54.02% （47/87） and 52.94% （36/68）, and the CC genotype of the two groups were 13.80% （12/87）, 11.76% （8/68）, respectively. The difference between the two groups was not statistically significant （χ2 =0. 24,P=0. 887）. The TT genotypes of rs1927914 were 33.33% （29/87） and 42.65% （29/68） in two groups, the CT genotypes of two groups were 52.87% （46/87） and 44.12% （30/68）, and the CC genotypes of two groups were 13.80% （12/87）, 13.23% （9/68）, respectively. The difference between the two groups was not statistically significant （χ2 = 1.49, P = 0. 475）. Rs7873784 gene locus of TLR 4 ： compared with wild homozygous genotype GG, the adjusted OR value of GC genotype was 0.90 （95%0CI：0.39-2.06, P=0.80）. Rs1927911 gene locus of TLR4 ： compared with wild homozygous genotype TT, the adjusted OR values of CT genotype and CC genotype were 1.00 （95GCI： 0.51- 1.97, P= 1.00） and 1.29 （95%CI： 0.46-3.66, P=0.63）, respectively. Rs1927914 gene locus of TLR4 ： compared with wild homozygous genotype TT, the adjusted OR values of CT genotype and CC genotype were 1.27 （ 95 G CI ： 0. 64- 2.51, P = 0. 49） and 1.24 （95 % CI： 0. 47- 3.27, P = 0. 67）, respectively. There was no significant difference between the 3 genotypes at different loci （P 〉 0.05 ）. Conclusion The polymorphisms of rs7873784, rs1927911 and rs1927914 in TLR4 gene are not associated with LEAD in T2DM-PTB patients.