广泛的药物性肝损伤及何首乌肝损伤致病基因研究

The incidence of drug induced liver injury (DILI) increased in recent years, and some cases with severe liver injury may present fulminant liver failure even death. In addition, it is not uncommon that patients with DILI have chronic course and progress to liver cirrhosis or induce autoimmune hepatitis. So doctors pay more and more attentions to DILI in clinic and how to prevent the occurrence of DILI also gradually referred to agenda. Since most DILI are idiosyncratic liver injury which has obvious individual differences and genetic predisposition, so research on genetic polymorphism of DILI has a realistic significance to preventing it to happen. Now researches on this respect in our country or abroad have preliminary progress, but few genes are clearly related to DILI, especially to traditional Chinese medicine, polygonum multiflorum, induced liver injury there has never been reported. This project propose to set 38 single nucleotide polymorphism (SNP) of 11 genes, widespread in all 10 patients with DILI or peculiar to patients with polygonum multiflorum induced liver injury, which were screened by entire exome sequencing of 10 patients ( 6 cases with polygonum multiflorum induced liver injury and 4 cases with clarithromycin induced liver injury) as candidate genes to verify the relations to extensive DILI and polygonum multiflorum induced liver injury using SNP loci PCR amplification and sequencing technology with large number of patients (200 cases with DILI including 50 case with polygonum multiflorum induced liver injury and 220 cases as healthy control).

药物性肝损伤（DILI）近年发病明显增加，且部分患者病情严重可为爆发性肝衰竭甚至死亡。而病程迁延转为慢性，进而发展为肝硬化或诱发自身免疫性肝炎者也屡见不鲜。因此，DILI在临床上越来越受到重视。而如何预防DILI也逐步提到日程上来。由于DILI大多为特异质性，有明显的个体差异和遗传倾向，所以， DILI遗传多态性研究对预防其发生有非常现实的意义。目前，国内外在此方面已有了初步研究，但能够明确与DILI相关基因寥寥无几，尤其中药何首乌引起肝损害相关基因国内外从未有报道。本课题拟以前期对10例患者血标本（何首乌6例、克拉霉素4例）进行的整个外显子组测序筛选出的10个样本中广泛存在的以及何首乌样本所特有的11个基因（38个位点）为备选基因，采用PCR及测序技术进行大样本（200例DILI患者，其中何首乌引起者50例。健康对照220例）验证与广泛的DILI及何首乌引起肝损伤的相关性。

药物性肝损伤（DILI）近年发病率明显增加，而如何预防DILI也逐步提到日程上来。DILI遗传多态性研究对预防其发生有非常现实的意义，因此，本研究目的为探索DILI及何首乌所引起肝损伤与遗传多态性的相关性。.本研究共收集570例广泛的药物性肝损伤患者临床资料，221例健康人和218例广泛的药物性肝损伤患者血标本，其中何首乌所致肝损伤患者50例。.对570例药物性肝损伤患者临床资料分析发现，药物性肝损伤患者以女性（66.8%）多见，涉及药物中中药（46.0%）占较高比例，其次是抗生素（8.4%）和心血管药物（5.1%）。肝损伤类型以肝细胞型（80.4%）为主。10%的病例可发展为慢性DILI，死亡率为1.9%。胆汁淤积型及混合型肝损伤易慢性化。.对收集的血标本，提取DNA后，采用时间飞行质谱技术检测439例血标本中97个单核苷酸多态性（single nucleotide polymorphism，SNP）位点的分布情况，对最小等位基因频率大于0.05的SNP进行统计学分析，发现以下基因上的SNP在组间的差异存在统计学意义：1. DRC1（rs3795958）、 MTHFR（rs1801133）、HLA-A（rs112069136、rs2231119）、HLA-B（rs1055348）、HLA-DRB1（rs111534875、rs202047044、rs150747106、rs78246137）等SNP可能与除何首乌外的其他药物所致肝损伤相关； 2. DRC1（rs3795958）、HLA-A（rs112069136）、HLA-B（rs1055348）、HLA-DRB1（rs111534875、rs202047044、rs150747106、rs78246137）等SNP可能与何首乌所致肝损伤相关； 3. HLA-B（rs1055348）在何首乌所致肝损伤组中的基因分布频率与对照组和其他药物所致肝损伤组之间的差异均存在统计学意义（P< 0.0001），具有何首乌特异性，与申请书中预实验结果一致。但rs1055348的临床意义尚未在ClinVar数据库中报道，其在何首乌所致肝损伤中的作用机制需要进一步的实验进行研究。上述研究结果可为后续研究药物性肝损伤及何首乌所致肝损伤分子发病机制及生物标记试剂盒的开发提供理论基础。