<i>TLR2</i>和<i>TLR4</i>基因多态性及启动子区CpG岛甲基化与抗结核药物性肝损伤的关系

李玥; 马烁; 刘越; 唐琴艳; 郭玉; 刘洋; 裴盛斐; 冯福民

doi:10.16462/j.cnki.zhjbkz.2025.02.015

TLR2和TLR4基因多态性及启动子区CpG岛甲基化与抗结核药物性肝损伤的关系

doi: 10.16462/j.cnki.zhjbkz.2025.02.015

李玥^{1, 2},
马烁¹,
刘越¹,
唐琴艳¹,
郭玉¹,
刘洋¹,
裴盛斐¹,
冯福民^1, ,

1.
华北理工大学公共卫生学院预防医学系, 河北省职业卫生与安全协同创新中心, 唐山 063210
2.
开滦总医院预防保健科, 唐山 063000

基金项目:

国家自然科学基金 81041096

详细信息

通讯作者:
冯福民, E-mail: fm_feng@sina.com

中图分类号: R183.3;Q756
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- 被引次数: 0
出版历程
- 收稿日期: 2023-11-20
- 修回日期: 2024-04-09
- 网络出版日期: 2025-03-08
- 刊出日期: 2025-02-10

Study on the relationship between TLR2 and TLR4 polymorphism and CpG island methylation and anti-tuberculosis drug-induced liver injury

LI Yue^{1, 2},
MA Shuo¹,
LIU Yue¹,
TANG Qinyan¹,
GUO Yu¹,
LIU Yang¹,
PEI Shengfei¹,
FENG Fumin^{1
, ,}

1.
School of Public Health, North China University of Science and Technology, Hebei Provincial Collaborative Innovation Center for Occupational Health and Safety, Tangshan 063210, China
2.
Department of Preventive Health Care, Kailuan General Hospital, Tangshan 063000, China

Funds:

National Natural Science Foundation of China 81041096

More Information

Corresponding author: FENG Fumin, E-mail: fm_feng@sina.com

摘要

摘要: 目的研究Toll样受体(Toll-like receptors, TLRs)家族成员中Toll样受体2(Toll-like receptor 2, TLR2)和Toll样受体4(Toll-like receptor 4, TLR4)基因的单核苷酸多态性(single nucleotide polymorphism, SNP)位点rs7656411、rs3804099、rs1927914、rs1927911及TLR2基因启动子区CpG岛(CpG island)甲基化与抗结核药物性肝损伤(anti-tuberculosis drug-related liver injury, ADLI)的关系。方法 2019年7月―2022年3月在招募的结核病患者中选择322名发生ADLI的患者和322名未发生ADLI的患者分别作为病例组和对照组, 按照性别、年龄和治疗方案进行1∶1匹配。通过甲基化特异性聚合酶链式反应(polymerase chain reaction, PCR)法和聚合酶链式反应-限制性片段长度多态性(PCR-restriction fragment length polymorphism, PCR-RFLP)技术检测基因多态性以及甲基化特征。结果 ADLI患者中TLR2基因rs3804099位点野生基因型(OR=0.55, 95% CI: 0.31~0.96)、CpG岛甲基化发生率高于对照组(OR=1.81, 95% CI: 1.07~3.06)。多变量分析表明TLR2基因rs3804099位点野生型TT与ADLI疾病进展相关, 野生型TT可能为ADLI的保护因素OR=0.53(95% CI: 0.38~0.75)。此外, TLR2基因CpG岛高甲基化与ADLI的发生风险独立相关[调整OR=1.86(95% CI: 1.05~3.28)]。rs3804099位点突变型和CpG岛高甲基化的交互作用增加了ADLI的发生风险, 为对照组的2.12倍(OR=2.12, 95% CI: 1.36~3.29)。结论 TLR2基因多态性和CpG岛甲基化与ADLI有关。TLR2基因启动子区CpG岛甲基化与其rs3804099位点基因多态性之间存在交互作用与ADLI的高风险有关。
- 基因多态性 /
- 抗结核药 /
- 肝损伤 /
- 甲基化
Abstract: Objective To investigate the relationship between single nucleotide polymorphisms rs7656411, rs3804099, rs1927914, rs1927911 in Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) genes, as well as CpG island methylation in the TLR2 gene promoter region, and anti-tuberculosis drug-induced liver injury (ADLI). Methods From July 2019 to March 2022, 322 tuberculosis patients with ADLI and 322 tuberculosis patients without ADLI were selected and matched 1∶1 based on gender, age, and treatment regimen. Gene polymorphisms and methylation patterns were detected using methylation-specific polymerase chain reaction(PCR) and PCR-restriction fragment length polymorphism(PCR-RFLP) technique. Results The wild-type genotype of TLR2 gene rs3804099 site (OR=0.55, 95% CI: 0.31-0.96) and CpG island methylation rate (OR=1.81, 95% CI: 1.07-3.06) were significantly higher in ADLI patients than in the control group (P < 0.05), with no statistical difference in the genotype frequency of other polymorphic sites between the two groups. Multivariate analysis showed that the wild-type TT of TLR2 gene rs3804099 site was associated with the progression of ADLI, and the wild-type TT may be a protective factor for ADLI OR=0.53 (95% CI: 0.38-0.75). In addition, high methylation of CpG island in TLR2 gene was independently associated with the risk of ADLI, with adjusted OR of 1.86 (95% CI: 1.05-3.28). The interaction between the mutation of rs3804099 site and high methylation of CpG island increased the risk of ADLI, being 2.12 times that of the control group (OR=2.12, 95% CI: 1.36-3.29). Conclusions There is an association between TLR2 gene polymorphism and CpG island methylation and ADLI. The interaction between CpG island methylation in the TLR2 gene promoter region and the gene polymorphism at rs3804099 is associated with a high risk of ADLI.
- Gene polymorphisms /
- Anti-tuberculosis drugs /
- Liver injury /
- Methylation

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图 1 各基因相关位点酶切电泳图

M: DNA marker; A: rs3804099位点; B: rs7656411位点; C: rs1927911位点; D: rs1927914位点。

Figure 1. Electrophoresis of enzyme digestion at each gene related site, based on enzyme after electrophoresis gel bands in the location and the number of rs3804099, rs7656411, rs1927911, rs1927914 loci genotyping

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图 2 TLR2基因MSP扩增产物电泳图

M: DNA marker; TLR2: Toll样受体2; 1, 2: 部分甲基化; 3, 4: 高甲基化; 5, 6: 低甲基化。

Figure 2. Electrophoresis of enzyme digestion at each gene related site, based on enzyme after electrophoresis gel bands in the location and the number of rs3804099, rs7656411, rs1927911, rs1927914 loci genotyping

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表 1 ADLI一般影响因素的单因素条件logistic回归分析结果

Table 1. Univariate logistic regression analysis of the general influencing factors (variables include marital status, occupation, smoking, education, Low density lipoprotein cholesterol, triglyceride, blood sugar) of antituberculosis drug-induced liver injury

影响因素	病例组	对照组	χ²/t值^①	P值^①	OR值(95% CI)^①	影响因素	病例组	对照组	χ²/t值^①	P值^①	OR值(95% CI)^①
婚姻状况						吸烟
未婚	96	91			1.00	否	238	254			1.00
已婚	226	231	0.322	0.570	0.28(0.14~1.52)	是	84	68	2.623	0.105	1.32(0.91~1.90)
职业						城乡居住地
其他	38	47			1.00	城市	90	81			1.00
工人	70	74	4.383	0.223	1.17(0.68~2.00)	乡村	232	242	0.147	0.701	1.07(0.74~1.54)
农民	155	160			3.20(0.74~4.94)	受教育情况
学生	59	41			2.78(0.99~3.19)	高中及以上	48	51			1.00
BMI/(kg·m^-2)						初中	116	110	0.278	0.870	2.12(0.70~2.80)
< 18.5	135	121			1.00	小学及以下	158	161			0.94(0.66~1.64)
≥18.5	187	201	1.754	0.185	0.83(0.61~1.14)	低密度脂蛋白胆固醇/(mmol·L^-1)	2.73±0.65	2.49±0.57	4.982	< 0.001
饮酒						三酰甘油/(mmol·L^-1)	1.62±0.98	1.49±0.89	1.762	0.079
否	220	257			1.00	血糖/(mmol·L^-1)	5.50±1.03	5.52±1.32	0.214	0.830
是	102	65	10.569	< 0.001	1.85(1.29~2.65)
注：ADLI, 抗结核药物性肝损伤。 ①病例组和对照组配对后统计分析得到的结果。

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表 2 两组TLR2基因和TLR4基因多态性分布比较

Table 2. The distribution of four SNP genotypes and alleles of TLR2 gene and TLR4 gene between the case group and the control group, and the statistical analysis results

基因	位点	类型	指标	病例组	对照组	χ²值^①	P值^①	OR值(95% CI)
TLR2	rs7656411	基因型	TT	157	148	3.247	0.072	0.56(0.33~1.05)
			GT	134	145	0.262	0.609	0.86(0.49~1.51)
			GG	31	29			1.00
		等位基因	T	448	441	0.178	0.673	1.05(0.75~1.47)
			G	196	203			1.00
	rs3804099	基因型	TT	115	138	5.261	0.022	0.55(0.31~0.96)
			CT	147	145	3.126	0.077	0.66(0.41~1.05)
			CC	60	39			1.00
		等位基因	T	377	421	6.377	0.012	0.75(0.64~0.88)
			C	267	223			1.00
TLR4	rs1927914	基因型	GG	146	136	2.453	0.117	0.63(0.36~1.22)
			GA	150	145	3.169	0.075	1.63(0.95~2.80)
			AA	26	41			1.00
		等位基因	G	442	417	2.184	0.139	1.19(0.67~2.11)
			A	202	227			1.00
	rs1927911	基因型	AA	145	137	2.782	0.095	0.85(0.37~1.09)
			AG	142	153	0.368	0.544	0.85(0.50~1.44)
			GG	35	32			1.00
		等位基因	A	432	427	0.087	0.768	1.04(0.77~1.40)
			G	212	217			1.00
注：TLR2, Toll样受体2; TLR4, Toll样受体4。 ①病例组和对照组配对后统计分析得到的结果。

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表 3 TLR2基因启动子区CpG岛甲基化情况在病例和对照中的分布

Table 3. Relationship between CpG island methylation in promoter region of TLR2 gene and anti-tuberculosis drug induced liver injury

对照组	病例组		χ²值	P值	OR值(95% CI)
对照组	高甲基化	低甲基化	χ²值	P值	OR值(95% CI)
高甲基化	10	21	4.898	0.026	1.81(1.07~3.06)
低甲基化	38	253
合计	48	274
注：TLR2, Toll样受体2。

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表 4 基于基因多态性ADLI影响因素的多因素条件logistic回归分析

Table 4. Multivariate logistic regression analysis was based on ADLI influencing factors (variables included alcohol consumption, low density lipoprotein cholesterol, TT type at rs3804099, hypermethylation)

因素	β值	s_x	Wald χ²值	P值	OR值(95% CI)
饮酒(对照组：不饮酒)	0.536	0.395	8.986	0.003	1.71(1.11~2.64)
低密度脂蛋白胆固醇	0.211	0.526	2.254	0.133	1.23(0.41~3.68)
rs3804099位点TT型(对照组：rs3804099位点CC型)	-0.634	0.629	4.789	0.029	0.53(0.38~0.75)
高甲基化(对照组：低甲基化)	0.621	0.747	7.396	0.007	1.86(1.05~3.28)
注：ADLI, 抗结核药物性肝损伤。

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表 5 TLR2基因启动子区CpG岛甲基化与其基因多态位点的交互作用分析

Table 5. Analysis of interaction between methylation of CpG island in promoter region of TLR2 gene and its polymorphic sites

基因多态性	甲基化水平	β值	χ²值	P值	OR值(95% CI)
TT	低				1.00
CC+CT	低	0.148	1.945	0.163	1.16(0.35~3.82)
TT	高	0.207	2.478	0.115	1.23(0.32~4.76)
CC+CT	高	0.751	5.631	0.018	2.12(1.36~3.29)
注：TLR2, Toll样受体2。

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参考文献(22)

[1]	李雨珊, 陆霓虹. 抗结核药物性肝损伤分子机制研究进展[J]. 中国医药科学, 2023, 13(14): 20-24. DOI: 10.20116/j.issn2095-0616.2023.14.04. Li YS, Lu NH. Research progress in the molecular mechanism of anti-tuberculosis drug-induced liver injury[J]. China Medicine and Pharmacy, 2023, 13(14): 20-24. DOI: 10.20116/j.issn2095-0616.2023.14.04.
[2]	Abbaspour F, Hasannezhad M, Khalili H, et al. Managing hepatotoxicity caused by anti-tuberculosis drugs: a comparative study of approaches[J]. Arch Iran Med, 2024, 27(3): 122-126. DOI: 10.34172/aim.2024.19.
[3]	Akkahadsee P, Sawangjit R, Phumart P, et al. Systematic review and network Meta-analysis of efficacy and safety of interventions for preventing anti-tuberculosis drug induced liver injury[J]. Sci Rep, 2023, 13(1): 19880. DOI: 10.1038/s41598-023-46565-3.
[4]	Kumar S, Duan QH, Wu RX, et al. Pathophysiological communication between hepatocytes and non-parenchymal cells in liver injury from NAFLD to liver fibrosis[J]. Adv Drug Deliv Rev, 2021, 176: 113869. DOI: 10.1016/j.addr.2021.113869.
[5]	Xu LY, Liu W, Bai FX, et al. Hepatic macrophage as a key player in fatty liver disease[J]. Front Immunol, 2021, 12: 708978. DOI: 10.3389/fimmu.2021.708978.
[6]	Chen DN, Xie WM, Lu YK, et al. Gene polymorphisms of TLR2 and TLR3 in HBV clearance and HBV-related hepatocellular carcinoma in a Chinese male population[J]. Int J Biol Markers, 2017, 32(2): e195-e201. DOI: 10.5301/jbm.5000238.
[7]	Neamatallah M, El-Bendary M, Elalfy H, et al. Impact of toll-like receptors 2( TLR2 ) and TLR4 gene variations on HCV susceptibility, response to treatment and development of hepatocellular carcinoma in cirrhotic HCV patients[J]. Immunol Invest, 2020, 49(4): 462-476. DOI: 10.1080/08820139.2019.1673772.
[8]	Elbrolosy AM, Elabd NS, ElGedawy GA, et al. Toll- like receptor 2 polymorphism and IL-6 profile in relation to disease progression in chronic HBV infection: a case control study in Egyptian patients[J]. Clin Exp Med, 2023, 23(1): 117-129. DOI: 10.1007/s10238-022-00792-6.
[9]	Shi GJ, Wang CX, Zhang P, et al. Donor polymorphisms of toll-like receptor 4 rs1927914 associated with the risk of hepatocellular carcinoma recurrence following liver transplantation[J]. Arch Med Res, 2017, 48(6): 553-560. DOI: 10.1016/j.arcmed.2017.11.011.
[10]	Wu DX, Li YH, Ren Q, et al. TANC1 methylation as a novel biomarker for the diagnosis of patients with anti-tuberculosis drug-induced liver injury[J]. Sci Rep, 2021, 11(1): 17423. DOI: 10.1038/s41598-021-96869-5.
[11]	许登高, 周扬, 潘庆杰. 哺乳动物CpG岛甲基化研究进展[J]. 青岛农业大学学报(自然科学版), 2012, 29(4): 252-260, 266. DOI: 10.3969/J.ISSN.1674-148X.2012.04.004. Xu DG, Zhou Y, Pan QJ. The methylation of CpG island in mammals[J]. Journal of Qingdao Agricultural University (Natural Science), 2012, 29(4): 252-260, 266. DOI: 10.3969/J.ISSN.1674-148X.2012.04.004.
[12]	Zhang D, Hao JQ, Hou RL, et al. The role of NAT2 polymorphism and methylation in anti-tuberculosis drug-induced liver injury in Mongolian tuberculosis patients[J]. J Clin Pharm Ther, 2020, 45(3): 561-569. DOI: 10.1111/jcpt.13097.
[13]	Bénichou C. Criteria of drug-induced liver disorders. Report of an international consensus meeting[J]. J Hepatol, 1990, 11(2): 272-276. DOI: 10.1016/0168-8278(90)90124-a.
[14]	Andersson T, Alfredsson L, Källberg H, et al. Calculating measures of biological interaction[J]. Eur J Epidemiol, 2005, 20(7): 575-579. DOI: 10.1007/s10654-005-7835-x.
[15]	Wu XZ, Dayanand KK, Thylur Puttalingaiah R, et al. Different TLR signaling pathways drive pathology in experimental cerebral malaria vs. malaria-driven liver and lung pathology[J]. J Leukoc Biol, 2023, 113(5): 471-488. DOI: 10.1093/jleuko/qiad021.
[16]	Brennan JJ, Gilmore TD. Evolutionary origins of toll-like receptor signaling[J]. Mol Biol Evol, 2018, 35(7): 1576-1587. DOI: 10.1093/molbev/msy050.
[17]	Xie JJ, Jiang SY, Shi MM, et al. The association between Toll-like receptor 2 single-nucleotide polymorphisms and hepatocellular carcinoma susceptibility[J]. BMC Cancer, 2012, 12: 57. DOI: 10.1186/1471-2407-12-57.
[18]	Elbrolosy AM, Elabd NS, ElGedawy GA, et al. Toll- like receptor 2 polymorphism and IL-6 profile in relation to disease progression in chronic HBV infection: a case control study in Egyptian patients[J]. Clin Exp Med, 2023, 23(1): 117-129. DOI: 10.1007/s10238-022-00792-6.
[19]	Deaton AM, Bird A. CpG islands and the regulation of transcription[J]. Genes Dev, 2011, 25(10): 1010-1022. DOI: 10.1101/gad.2037511.
[20]	Harahap WA, Sudji IR, Nindrea RD. BRCA1 promoter methylation and clinicopathological characteristics in sporadic breast cancer patients in Indonesia[J]. Asian Pac J Cancer Prev, 2018, 19(9): 2643-2649. DOI: 10.22034/APJCP.2018.19.9.2643.
[21]	Shen LH, Zhang B, Sun SF, et al. Methylation of cytochrome p450 2E1 promoter induced by low dosage of isoniazid[J]. Environ Toxicol Pharmacol, 2013, 36(1): 149-151. DOI: 10.1016/j.etap.2013.03.016.
[22]	Zhang B, Sun SF, Shen LH, et al. DNA methylation in the rat livers induced by low dosage isoniazid treatment[J]. Environ Toxicol Pharmacol, 2011, 32(3): 486-490. DOI: 10.1016/j.etap.2011.07.001.