肝豆状核变性基因高频突变位点与中医证型关系的探讨

目的：探讨肝豆状核变性（HLD）ATPTB基因高频突变位点Arg778Leu、Thr935Met、Pr0992Leu点突变与中医证型的关系。方法：选择142例无血缘关系的HLD确诊患者，采用TouchdownPCR（TD—PCR）扩增ATP7B基因的第8、12、13号外显子，并对PCR扩增产物予以限制性内切酶酶切检测第778、935、992密码子的基因突变，对检测的阳性结果进行DNA测序验证。同时对以上142例HLD患者进行中医辨证分型，分析基因突变类型与中医证型的关系。结果：142例ttLD患者中48．59Y0（69／142）的患者在第8外显子检出hrg778Leu突变，5．63％（8／142）的患者在第12外显子检出Thr935Met杂合突变，24．65％（35／142）的患者在第13外显子检出Pro992Leu突变。发生hrg778Leu突变的患者中，肝肾阴虚型32例，气血两虚型13例，湿热内蕴型21例：发生Pr0992Leu突变的患者中，肝肾阴虚型12例，气血两虚型9例，湿热内蕴型11例。结论：hrg778Leu突变类型与肝肾阴虚型存在相关性（P=-0．03）。Pro992Leu突变类型与各中医证型无相关性（P〉0．05）。

Exploration on the Relationship between TCM Patterns and High Frequency Mutated Gene Locus in the Patients with Hepatolenticular Degeneration

Objective： To discuss the relationship between TCM patterns and the mutations of gene locus including Arg 778Leu, Thr935Met and Pro992Leu located at ATP7B gene of Chinese patients suffering from hepatolenticular degeneration（HLD）. Method： All 142 patients with HLD were chosen, the exon 8, 12 and 13 located at ATP7B gene was amplified with Touchdown PCR （TD-PCR）, gene mutation of the codon 778, 935 and 992 was tested through amplification product of PCR dissected with restriction enzyme, the positive results were tested with DNA sequencing. Above 142 patients were differentiated into patterns, the relationship between gene mutation and TCM patterns was analyzed. Result： Among 142 HLD patients, 48.59%（69/142） of the patients were tested with Arg778Leu mutation in the exon 8, 5.63% （8/142） of the patients were detected with Thr935Met heterozygous mutation in the exon 12; 24.65%（35/142） of the patients were tested with Pro992Leu mutation. Among the patients with Arg778Leu mutation, there were 32 cases of liver-kidney Yin deficiency pattern, 13 cases of dual deficiency of Qi-blood and 21 cases of internal accumulation of dampness-heat; among the patients with Pro992Leu mutation, there were 12 cases of liver-kidney Yin deficiency pattern, nine cases of dual deficiency of Qi-blood and 11 cases of internal accumulation of dampness-heat. Conclusion： The types of Arg778Leu mutation are related to liver-kidney Yin deficiency pattern （P=-0.03）. The types of Pro992Leu mutation show no relativity to different TCM patterns （P〉0.05）.