中国病理生理杂志2012，Vol.28Issue(7)：1308-1313,6.DOI:10.3969/j.issn.1000-4718.2012.07.029
HRM技术在瘦素基因启动子多态性与肝硬化发生相互关系研究中的应用
Application of PCR-HRM in the study of relationship between leptin gene promoter polymorphisms and liver cirrhosis
摘要
Abstract
AIM: To analyze the feasibility of PCR — high — resolution melting curve analysis ( HRM ) for detecting the site mutations of C2549 and G2548 in leptin gene promoter from the patients with liver cirrhosis, and to explore the relevance between mutant genotypes and physiological and biochemical indexes in liver cirrhosis patients. METHODS: Compared with the method of PCR — restriction fragment length polymorphism ( RFLP ), the present research used the method of PCR — HRM to analyze the site mutations of C2549 and G2548 in leptin gene promoter in control group ( n = 100 ) and liver cirrhosis group ( n = 100 ). The physiological and biochemical indexes of the patients were also detected and compared. RESULTS: Leptin gene promoter polymorphism was detected using PCR — HRM with effectiveness, high flux and accuracy. Preliminary results showed that the main mutation of the patients with liver cirrhosis was in C2549 site, but not found in G2548 site. Leptin, free leptin index ( FLI ), fasting insulin ( FINS ) and insulin resistance index estimated by homeostatic model assessment ( HOMA — IR ) in liver cirrhosis group were higher than those in control group. Insulin sensitivity index ( ISI ) and soluble leptin receptor ( sOB — R ) in liver cirrhosis group were lower than those in control group with significant difference except leptin level. Meanwhile, FLI showed positively correlated with FINS and HOMA — IR ( r = 0.45, r = 0. 53 , P < 0. 05 ), and negatively with ISI ( r = - 0. 34, P < 0. 05 ). In the patients with liver cirrhosis, C2549A heterozygous mutation was predominant. The indexes of HOMI — IR, leptin, sOB — R and FLI of C2549A homozygotes and heterozygotes were higher than those of the wildtypes, which showed significant difference except leptin and sOB - R levels ( P <0. 05 ). CONCLUSION: PCR — HRM can be more accurate for identifying leptin promoter polymorphism. The increase in the frequency of C2549A mutation may be closely related with liver cirrhosis. Existence of hyperinsulinemia and insulin resistance may be correlated with leptin level in the patients with liver cirrhosis.关键词
高分辨率熔解曲线分析/瘦素/基因启动子/基因多态性/肝硬化Key words
High - resolution melting curve analysis/ Leptin/ Promoter, genetic/ Polymorphism, genetic/ Liver cirrhosis分类
医药卫生引用本文复制引用
李善高,曹海军,刘军,徐国萍,孙明洪,朱孝鸿,吕宾,孟立娜..HRM技术在瘦素基因启动子多态性与肝硬化发生相互关系研究中的应用[J].中国病理生理杂志,2012,28(7):1308-1313,6.基金项目
浙江省自然科学基金资助项目(No.Y2100379) （No.Y2100379）
