rs3804100、rs3804099及rs2295080基因多态性与黑龙江省2型糖尿病患者并发肺结核关系的研究

doi:10.3969/j.issn.2095-3755.2020.01.011

摘要/Abstract

摘要：

目的探究Toll样受体2(Toll-like receptor 2,TLR2)rs3804100、rs3804099位点和哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)rs2295080位点基因多态性与2型糖尿病并发肺结核的关系。方法选择2017年1—9月黑龙江省传染病防治院收治的300例2型糖尿病并发肺结核患者作为观察组,以及同时期在哈尔滨医科大学附属第四医院内分泌科就诊的300例2型糖尿病患者作为对照组。所有患者收集血液提取DNA,用PCR技术对TLR2的rs3804100、rs3804099位点及mTOR的rs2295080位点进行基因多态性检测,并对这3个基因位点多态性与2型糖尿病并发肺结核的关系进行单因素和logistic多因素分析。结果单因素分析结果显示,TLR2的rs3804100位点TT、TC、CC(T:胸腺嘧啶;C:胞嘧啶)基因型在观察组和对照组分布构成比分别为51.3%(154/300)、25.7%(77/300)、23.0%(69/300)和47.0%(141/300)、31.3%(94/300)、21.7%(65/300),差异无统计学意义(χ²=2.382,P=0.304);TLR2的rs3804099位点TT、TC、CC基因型在观察组和对照组分布构成比分别为53.0%(159/300)、29.7%(89/300)、17.3%(52/300)和54.3%(163/300)、33.0%(99/300)、12.7%(38/300),差异无统计学意义(χ²=2.759,P=0.252);mTOR的rs2295080位点TT、TG、GG(G:鸟嘌呤)基因型在观察组和对照组分布构成比分别为26.7%(80/300)、67.3%(202/300)、6.0%(18/300)和24.3%(73/300)、66.0%(198/300)、9.7%(29/300),差异无统计学意义(χ²=2.935,P=0.231)。logistic多因素分析结果显示,TLR2的rs3804100位点TC、CC基因型的OR(95%CI)值分别为1.261(0.591~2.689)和1.284(0.542~3.042),P值分别为0.549和0.569;TLR2的rs3804099位点TC、CC基因型的OR(95%CI)值分别为0.752(0.461~1.227)和0.729(0.430~1.235),P值分别为0.254和0.240;mTOR的rs2295080位点TG、GG基因型的OR(95%CI)值分别为1.789(0.890~3.596)和1.603(0.839~3.063),P值分别为0.103和0.153。结论 TLR2基因rs3804100、rs3804099位点及mTOR的rs2295080位点多态性与黑龙江省2型糖尿病并发肺结核均无相关性。

关键词: 结核,肺, 糖尿病,2型, 糖尿病并发症, 扩增片段长度多态性分析, 因素分析,统计学

Abstract:

Objective To investigate the relationship between genetic polymorphisms rs3804100, rs3804099 in Toll-like receptor 2 (TLR2), rs2295080 in mammalian target of rapamycin (mTOR) and type 2 diabetes mellitus complicated with pulmonary tuberculosis (PTB). Methods From January 2017 to September 2017, 300 patients with PTB complicated by type 2 diabetes in Infectious Diseases Hospital of Heilongjiang Province were selected as observation group, and 300 patients with type 2 diabetes who admitted to the endocrinology department of Fourth Affiliated Hospital of Harbin Medical University were included as control group at the same time. DNA was extracted from the blood of all patients, and genetic polymorphisms (rs3804100, rs3804099 in TLR2, rs2295080 in mTOR) was detected by PCR technology. The relationship between the polymorphisms in these three gene loci and type 2 diabetes complicated with PTB was performed by univariate and multivariate logistic analysis. Results Univariate analysis showed that TT, TC and CC (T, thymine; C, cytosine) genotypes distribution composition ratios in TLR2 rs3804100 were 51.3% (154/300), 25.7% (77/300), and 23.0% (69/300) in the observation group; and 47.0% (141/300), 31.3% (94/300), and 21.7% (65/300) in the control group, respectively, without the statistically significant difference between these two groups (χ²=2.382, P=0.304). The TT, TC and CC genotypes distribution composition ratios in TLR2 rs3804099 were 53.0% (159/300), 29.7% (89/300), and 17.3% (52/300) in the observation group; and 54.3% (163/300), 33.0% (99/300), and 12.7% (38/300) in the control group, respectively, without the statistically significant difference (χ²=2.759, P=0.252). The TT, TG and GG (G, guanine) genotypes distribution composition ratios in mTOR rs2295080 were 26.7% (80/300), 67.3% (202/300), and 6.0% (18/300) in the observation group; and 24.3% (73/300), 66.0% (198/300), and 9.7% (29/300) in the control group, respectively, without the statistically significant difference (χ²=2.935, P=0.231). The results of logistic multivariate analysis revealed that the OR (95%CI) values of TC, CC genotypes in TLR2 rs3804100 were 1.261 (0.591-2.689) and 1.284 (0.542-3.042), respectively; P values were 0.549 and 0.569, respectively. The OR (95%CI) values of TC, CC genotypes in TLR2 rs3804099 were 0.752 (0.461-1.227) and 0.729 (0.430-1.235), respectively; P values were 0.254 and 0.240, respectively. The OR (95%CI) values of TG, GG genotypes in mTOR rs2295080 were 1.789 (0.890-3.596) and 1.603 (0.839-3.063), respectively; P values were 0.103 and 0.15, respectively. Conclusion The genetic polymorphisms rs3804100, rs3804099 in TLR2 and polymorphisms rs2295080 in mTOR are not associated with type 2 diabetes mellitus complicated with PTB in Heilongjiang Province.

Key words: Tuberculosis, pulmonary, Diabetes mellitus, type 2, Diabetes complications, Amplified fragment length polymorphism analysis, Factor analysis, statistical

陈海军, 薄璐, 郭鑫, 张堃, 赫丽杰, 李强, 李雨泽. rs3804100、rs3804099及rs2295080基因多态性与黑龙江省2型糖尿病患者并发肺结核关系的研究[J]. 结核病与肺部健康杂志, 2020, 9(1): 49-54. doi: 10.3969/j.issn.2095-3755.2020.01.011

CHEN Hai-jun, BO Lu, GUO Xin, ZHANG Kun, HAO Li-jie, LI Qiang, LI Yu-ze. Relationship between genetic polymorphisms rs3804100, rs3804099 and rs2295080 and type 2 diabetes mellitus complicated with pulmonary tuberculosis in Heilongjiang Province[J]. Journal of Tuberculosis and Lung Health, 2020, 9(1): 49-54. doi: 10.3969/j.issn.2095-3755.2020.01.011

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试剂	体积(μl)	反应条件
2×通用PCR预混液	2.500	95℃ 10min
40×探针引物混合物	0.125	95℃ 15s,60℃ 40s; 42个循环
双蒸水	2.375	50℃ 30s
合计	5.000

序号	变量	赋值
X1	rs3804100位点	0=TT;1=TC;2=CC
X2	rs3804099位点	0=TT;1=TC;2=CC
X3	rs2295080位点	0=TT;1=TG;2=GG
X4	性别	0=男;1=女
X5	年龄	0=≤60岁;1=>60岁

单核苷酸多态性组别	胸腺嘧啶		胞嘧啶/鸟嘌呤		χ²值	P值
单核苷酸多态性组别	等位基因频数	等位基因频率(%)	等位基因频数	等位基因频率(%)	χ²值	P值
rs3804100位点					0.291	0.590
对照组	376	62.7	224^a	37.3^a
观察组	385	64.2	215^a	35.8^a
rs3804099位点					1.270	0.260
对照组	425	70.8	175^a	29.2^a
观察组	407	67.8	193^a	32.2^a
rs2295080位点					1.115	0.291
对照组	344	57.3	256^b	42.7^b
观察组	362	60.3	238^b	39.7^b

因素	β值	s$\bar{x}$值	Wald χ²值	P值	OR值	95%CI值
因素	β值	s$\bar{x}$值	Wald χ²值	P值	OR值	下限	上限
rs3804100位点
TT	-	-	-	-		1.000
TC	-0.011	0.217	0.003	0.549	1.261	0.591	2.689
CC	-0.328	0.241	1.850	0.569	1.284	0.542	3.042
rs3804099位点
TT	-	-	-	-		1.000
TC	-0.285	0.250	1.302	0.254	0.752	0.461	1.227
CC	-0.316	0.269	1.380	0.240	0.729	0.430	1.235
rs2295080位点
TT	-	-	-	-		1.000
TG	0.581	0.356	2.662	0.103	1.789	0.890	3.596
GG	0.472	0.330	2.043	0.153	1.603	0.839	3.063
性别
男	-	-	-	-		1.000
女	-0.581	0.172	11.428	0.001	0.559	0.399	0.783
年龄组(岁)
<50	-	-	-	-		1.000
50~	1.119	0.274	19.184	0.000	3.318	1.940	5.675
60~	0.409	0.268	2.331	0.127	1.505	0.890	2.544
≥70	0.267	0.283	0.894	0.344	1.306	0.751	2.274