细胞因子基因多态性与肺结核并发糖尿病易感性的相关性研究

doi:10.3969/j.issn.1000-6621.2018.10.005

摘要/Abstract

摘要： 目的

探索细胞因子γ-干扰素(IFN-γ)+874A/T位点、白细胞介素(IL)-10 -1082G/A位点和-592A/C位点基因多态性与肺结核并发糖尿病(pulmonary tuberculosis and diabetic mellitus,PTB-DM)易感性的关系,以及遗传危险因素在PTB-DM发病机制中的作用,为PTB-DM高危人群基因多态性筛查提供理论依据。

方法

采用病例对照研究方法,纳入江苏省2013年4月1日至2014年3月31日3个市(县)142例PTB-DM患者(PTB-DM组),同期选取诊断为结核病的患者147例(TB组)和糖尿病患者141例(DM组)作为对照。通过结构式问卷调查获取研究对象社会人口学信息,采集患者血样并提取DNA,采用聚合酶链式反应并限制性片段长度多态性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLF)方法检测IFN-γ+874A/T位点、IL-10 -1082G/A位点和-592A/C位点基因多态性。采用EpiData和SPSS 16.0统计软件录入与分析数据,运用单因素方差分析和多因素logistic回归分析IFN-γ+874A/T位点、IL-10 -1082G/A位点和-592A/C位点基因多态性与PTB-DM易感性的相关性。

结果

IFN-γ+874A/T、IL-10 -592A/C、IL-10 -1082G/A位点经PCR扩增和基因多态性位点分型鉴定,分别获得等位基因A和T、A和C、G和A,以及基因型AA、AT、TT;AA、AC、CC;GA、AA。基因多态性检测结果显示,仅PTB-DM组IL-10 -1082G/A位点A等位基因频率(92.61%,263/284)高于DM组(87.59%,247/282)(χ²=3.995,P=0.046)、AA基因型频率(85.21%,121/142)高于DM组(75.18%,106/141)[校正OR(aOR)=1.970;95%CI:1.066~3.643;P=0.030]。IL-10 -592和-1082两位点间存在连锁不平衡(D'=0.959,r²=0.181,P<0.001),PTB-DM患者的IL-10 C-A单倍型频率(25.90%)明显高于DM组患者(17.60%)(OR=1.590,95%CI=1.056~2.396,P=0.026)。

结论

IL-10 -1082位点的AA基因型可能与PTB-DM易感性有关,为疾病共患的危险因素;IL-10单倍型与PTB-DM易感性存在相关性,其C-A单倍型可能使DM患者共患TB的风险上升。而IL-10 -592A/C位点和IFN-γ +874A/T位点可能与DM患者对TB的易感性、或者TB患者对DM的易感性无关。

关键词: 结核, 肺, 糖尿病, 共病现象, 细胞因子类, 扩增片段长度多态性分析, 疾病易感性, 遗传相关性研究

Abstract: Objective

This study aims to reveal the associations among IFN-γ +874 A/T, IL-10 -592 A/C, -1082 G/A SNPs and risk of diabetic pulmonary tuberculosis, and consequently provide further evidence for research on pathogenesis of diabetic pulmonary tuberculosis (PTB-DM) and the screening for diabetic pulmonary tuberculosis patients.

Methods

A case-control study based on the screening of DM in PTB patients (142 cases) was carried out in 3 counties in Jiangsu Province from 1 April, 2013 to 31 March, 2014. Cases in this study were all diagnosed diabetic pulmonary tuberculosis patients, whereas control groups were PTB patients (147 cases) and DM patients (141 cases) matching with gender and age. Social demographic information was collected by a questionnaire and DNA was extracted from the blood sample collected.The polymorphisms of IFN-γ +874 A/T, IL-10 -592 A/C and -1082 G/A were detected by PCR-RFLF methods and followed by direct sequencing. Statistical analyses were conducted using the EpiData and SPSS software version 16.0. Pearson chi-square and logistic regression were used to estimate the associations among IFN-γ+874 A/T, IL-10 -592 A/C, -1082 G/A SNPs and risk of diabetic pulmonary tuberculosis.

Results

In the study on IL-10 -1082 G/A, statistical differences were discovered between PTB-DM patients and DM controls (A allele, 92.61% (263/284) versus 87.59% (247/282)(χ²=3.995, P=0.046); AA genotype, 85.21% (121/142) versus 75.18% (106/141). The AA genotype of IL-10 had a 1.970 fold increased risk for diabetic pulmonary tuberculosis individuals (OR=1.970, 95%CI=1.066-3.643, respectively). Linkage disequilibrium was found between IL-10 -592 and IL-10 -1082 (D'=0.959,r²=0.181,P<0.001) and statistical significance was also found in IL-10 -592C/-1082A haplotype between diabetic pulmonary tuberculosis patients and DM controls (25.90% versus 17.60%, P=0.026); IL-10 -592C/-1082A haplotype had a 1.590 fold increased risk for diabetic pulmonary tuberculosis individuals (OR=1.590, 95%CI=1.056-2.396, respectively).

Conclusion

Our results indicated an association between IL-10 -1082G/A polymorphism and susceptibility to diabetic pulmonary tuberculosis. Novel risk haplotype of IL-10 has been identified, of which -592C/-1082A haplotype was found to be strongly linked to diabetic pulmonary tuberculosis susceptibility. However, no association was found between IL-10 -592 or IFN-γ +874 and the risk of diabetic pulmonary tuberculosis.

Key words: Tuberculosis, pulmonary, Diabetes mellitus, Comorbidity, Cytokines, Amplified fragment length polymorphism analysis, Disease susceptibility, Genetic association studies

董斯佳,袁立,陈诚,蒋伟利,赵琦. 细胞因子基因多态性与肺结核并发糖尿病易感性的相关性研究[J]. 中国防痨杂志, 2018, 40(10): 1051-1059. doi: 10.3969/j.issn.1000-6621.2018.10.005

Si-jia DONG,Li YUAN,Cheng CHEN,Wei-li JIANG,Qi ZHAO. Association of cytokine genetic polymorphisms with susceptibility to diabetic pulmonary tuberculosis[J]. Chinese Journal of Antituberculosis, 2018, 40(10): 1051-1059. doi: 10.3969/j.issn.1000-6621.2018.10.005

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各位点基因型指标	PTB-DM组(142例)			&#x003c7;²值	P值	PTB组(147例)			&#x003c7;²值	P值	DM组(141例)			&#x003c7;²值	P值
IFN-&#x003b3;+874A/T	AA	AT	TT	0.110	0.740	AA	AT	TT	6.700	0.009	AA	AT	TT	0.018	0.894
观察频数 (频率,%)	107 (75.35)	32 (22.54)	3 (2.11)			118 (80.82)	23 (15.75)	5 (3.43)			108 (76.60)	31 (21.99)	2 (1.41)
预期频数 (频率,%)	106 (74.65)	33 (23.24)	3 (2.11)			115 (78.77)	29 (19.86)	2 (1.37)			108 (76.60)	31 (21.99)	2 (1.41)
IL-10 -592A/C	AA	AC	CC	0.218	0.640	AA	AC	CC	0.548	0.459	AA	AC	CC	0.100	0.572
观察频数 (频率,%)	63 (44.37)	65 (45.77)	14 (9.86)			60 (40.82)	71 (38.30)	16 (10.88)			73 (53.28)	53 (38.69)	11 (8.03)
预期频数 (频率,%)	64 (45.07)	63 (44.37)	15 (10.56)			62 (42.18)	67 (45.58)	18 (12.24)			72 (52.94)	54 (39.71)	10 (7.35)
IL-10 -1082G/A	GG	GA	AA	0.905	0.341	GG	GA	AA	2.193	0.139	GG	GA	AA	2.831	0.092
观察频数 (频率,%)	0 (0.00)	21 (14.79)	121 (85.21)			0 (0.00)	32 (21.77)	115 (78.23)			0 (0.00)	35 (24.82)	106 (75.18)
预期频数 (频率,%)	1 (0.70)	19 (13.38)	122 (85.92)			1 (0.68)	29 (19.73)	117 (79.59)			2 (1.41)	31 (21.99)	108 (76.60)

等位基因	PTB-DM组(142例)		PTB组(147例)		DM组(142例)		&#x003c7;²值^a	P值^a	&#x003c7;²值^b	P值^b	&#x003c7;²值^c	P值^c
等位基因	频数	频率(%)	频数	频率(%)	频数	频率(%)	&#x003c7;²值^a	P值^a	&#x003c7;²值^b	P值^b	&#x003c7;²值^c	P值^c
IFN-&#x003b3; +874A/T							0.576	0.750	0.576	0.448	0.118	0.731
A	246	86.62	259	88.70	247	87.59
T	38	13.38	33	11.30	35	12.41
IL-10 -592A/C							4.011	0.135	0.337	0.561	1.914	0.167
A	191	67.25	191	64.97	199	72.63
C	93	32.75	103	35.03	75	27.37
IL-10 -1082G/A							4.084	0.130	2.113	0.146	3.995	0.046
G	21	7.39	32	10.88	35	12.41
A	263	92.61	262	89.12	247	87.59

基因型	PTB-DM组(142例)		PTB组(147例)		DM组(141例)		&#x003c7;²值	P值
基因型	频数	频率(%)	频数	频率(%)	频数	频率(%)	&#x003c7;²值	P值
IFN-&#x003b3; +874A/T							3.633	0.458
AA	107	75.35	118	80.82	108	76.59
AT	32	22.54	23	15.75	31	21.99
TT	3	2.11	5	3.42	2	1.42
IL-10 -592A/C							4.706	0.319
AA	63	44.37	60	40.82	73	53.28
AC	65	45.77	71	48.30	53	38.69
CC	14	9.86	16	10.88	11	8.03
IL-10 -1082G/A							4.609	0.100
GG	0	0.00	0	0.00	0	0.00
GA	21	14.79	32	21.77	35	24.82
AA	121	85.21	115	78.23	106	75.18

单倍型	PTB-DM组(142例)		PTB组(147例)		OR(95%CI)值	P值
单倍型	频数	频率(%)	频数	频率(%)	OR(95%CI)值	P值
A-A	189.51	66.70	191.00	65.00	1.990(0.778~1.552)	0.592
C-A	73.49	25.90	71.00	24.20	1.104(0.758~1.610)	0.606
C-G	19.51	6.90	32.00	10.90	0.607(0.337~1.094)	0.094
A-G	-	-	-	-	-	-

单倍型	PTB-DM组(142例)		DM组(147例)		aOR(95%CI)值	P值
单倍型	频数	频率(%)	频数	频率(%)	aOR(95%CI)值	P值
A-A	189.51	66.70	191.17	70.00	0.797(0.554~1.148)	0.233
C-A	73.49	25.90	48.29	17.60	1.590(1.056~2.396)	0.026
C-G	19.51	6.90	26.71	9.70	0.666(0.362~1.226)	0.189
A-G	-	-	-	-	-	-