线粒体DNA控制区变异与瘢痕疙瘩的相关性研究

doi:10.35541/cjd.20201021

摘要/Abstract

摘要： 【摘要】目的探讨线粒体DNA（mtDNA）控制区D环（D-loop）的突变与瘢痕疙瘩的相关性。方法收集2016—2019年昆明医科大学第一附属医院皮肤科门诊瘢痕疙瘩患者216例，提取患者外周血总DNA及25例患者的瘢痕疙瘩组织、瘢痕疙瘩旁正常组织的总DNA。以云南大学附属医院体检中心无瘢痕疙瘩的健康体检者299例外周血样本数据作为对照。对mtDNA D-loop区进行PCR扩增及Sanger测序，并与修正后的剑桥参考序列（rCRS）比对，获得每个样本的突变位点。根据Phylotree-mtDNA tree Build 17，对2组人群进行单倍型类群划分。比较瘢痕疙瘩组织、瘢痕疙瘩旁正常组织及自身外周血mtDNA的D-loop区突变。使用network 5.0软件制作中介网络图，采用二元logistic回归分析单倍型类群频率与瘢痕疙瘩发病的相关性，χ2、t及t′检验分析临床数据。结果 216例瘢痕疙瘩患者mtDNA可划分为10个单倍型类群：A、B、D、R9、G、M*、M7、M8、M9、N9，其中R9分布频率最高（21.3%，46/216），M9分布频率最低（0.9%，2/216）。瘢痕疙瘩患者的单倍型类群M7和N9的分布频率均显著低于对照人群（P = 0.040，OR = 0.248，95% CI：0.066 ~ 0.937；P = 0.048，OR = 0.191，95% CI：0.037 ~ 0.986）。中介网络图显示，瘢痕疙瘩患者和对照人群的单倍型类群M7具有不同的分布模式。单倍型类群M7患者的发病部位数比非M7患者少（P = 0.000 1），且发病年龄比非M7患者小（P = 0.045）。结论单倍型类群M7与瘢痕疙瘩的发生具有相关性，可能是瘢痕疙瘩发生的潜在保护性因素。

关键词: 瘢痕疙瘩, DNA, 线粒体, D环, 单倍型类群, 体细胞突变

Abstract: 【Abstract】 Objective To investigate the correlation between variations in mitochondrial DNA （mtDNA） control region （D-loop） and keloids. Methods A total of 216 patients with keloids were collected from Department of Dermatology, the First Affiliated Hospital of Kunming Medical University from 2016 to 2019. Total DNA was extracted from peripheral blood samples of all the patients, as well as keloid tissues and perilesional normal skin tissues of 25 patients with keloids. Peripheral blood samples were collected from 299 health checkup examinees without keloids in Health Examination Center, the Affiliated Hospital of Yunnan University, who served as controls. PCR amplification and Sanger sequencing were performed on the mtDNA D-loop region, and mutation sites in each sample were analyzed by comparisons with the revised Cambridge Reference Sequence （rCRS）. Haplogroups were assigned in the 2 groups by using Phylotree-mtDNA tree Build 17. Mutations in the mtDNA D-loop region were compared among keloid tissues, perilesional normal skin tissues and peripheral blood samples. A median-joining network was constructed via network 5.0 software. Binary logistic regression analysis was performed to investigate the correlation between haplogroup frequencies and the occurrence of keloids, and chi-square, t and t′ tests were used to analyze clinical data. Results Among the 216 patients with keloids, variations in mtDNA D-loop region were classified into 10 haplogroups, including A, B, D, R9, G, M*, M7, M8, M9 and N9, with the haplogroups R9 and M9 showing the highest （21.3%, 46/216） and lowest （0.9%, 2/216） frequencies respectively. The frequencies of haplogroups M7 （P = 0.040, OR = 0.248, 95% CI: 0.066 - 0.937） and N9 （P = 0.048, OR = 0.191, 95% CI: 0.037 - 0.986） were significantly lower in the patients with keloids than in the controls. The median-joining network plot showed that the distribution pattern of the haplogroup M7 differed between the patients with keloids and controls. Significantly less number of lesional sites and younger age of onset were observed in the patients with haplogroup M7 compared with those with non-M7 haplogroups （P = 0.000 1, 0.045, respectively）. Conclusion The haplogroup M7 is correlated with the occurrence of keloids, and may be a potential protective factor for keloid formation.

Key words: Keloid, DNA, mitochondrial, D-loop, Haplogroup, Somatic mutation

郭一俨周太成李改赢罗璇王睿祺麻艺群蒋艳汤諹. 线粒体DNA控制区变异与瘢痕疙瘩的相关性研究[J]. 中华皮肤科杂志, 2021, 54(5): 421-427.doi:10.35541/cjd.20201021

Guo Yiyan, Zhou Taicheng, Li Gaiying, Luo Xuan, Wang Ruiqi, Ma Yiqun, Jiang Yan, Tang Yang. Correlation between mitochondrial DNA control region variations and keloid formation[J]. Chinese Journal of Dermatology, 2021, 54(5): 421-427.doi:10.35541/cjd.20201021

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