ApoAI、B基因多态性与非创伤性股骨头坏死证候相关性研究

目的:研究载脂蛋白AI、B(ApoAI、B)基因多态性与非创伤性股骨头坏死(nontraumatic osteonecrosis of femoral head,NONFH)中医证候的相关性。方法:采用聚合酶链式反应(PCR)和限制性片段长度多态性(RFLP)对143例NON-FH患者和92例正常人分别进行ApoAI基因启动子-75bp位点、内含子+83bp位点,ApoB基因EcoRI、XbaI的多态性研究,采用聚合酶链反应(PCR)和琼脂糖凝胶电泳分析ApoB基因3’端可变数目串联重复序列(3’-VNTR);对NONFH患者按痰瘀阻络、经脉痹阻和肝肾亏虚3个证型进行中医辨证,用SAS软件分析基因型、等位基因在NONFH证候分型出现频率。结果:ApoAI启动子-75bp基因位点,经脉痹阻证和肝肾亏虚证的A/A基因型频率均高于对照组(P0.05),肝肾亏虚证的G/A基因型频率显著低于对照组(P0.01)、痰瘀阻络证和经脉痹阻证(P0.05),而G/G基因型频率明显高于经脉痹阻证(P0.05);等位基因比较中,经脉痹阻证G等位基因频率显著低于其他各组(P0.05)。ApoAI启动子-内含子中,与痰瘀阻络相比,经脉痹阻证组的G/G-C/C、G/A-C/C基因型频率均显著降低(P0.05);肝肾亏虚证组G/A-C/C基因型频率显著降低(P0.05);而且肝肾亏虚证组G/A-C/C基因型频率显著低于经脉痹阻证组。ApoB基因XbaI基因型中,经脉痹阻证组X+X-基因型频率明显高于对照组(P0.05),但X+、X-等位基因频率无明显差异(P0.05)。在ApoAI基因内含子+83bp位点、ApoB基因EcoRI基因型、3’-VNTR基因型频率、等位基因频率在各证型及对照组间分布无明显差异(P0.05)。结论:ApoAI基因启动子-75bp位点多态性、启动子——内含子位点多态性及ApoB基因XbaI基因型与NONFH证候分型有一定相关性;ApoAI基因内含子+83bp位点、ApoB基因EcoRI、3’-VNTR多态性可能与NONFH的证候发生、发展无相关关系。

Study on the Correlation between Apolipoprotein AI and B Gene Polymorphism and the Syndrome of Nontraumatic Osteonecrosis of Femoral Head

Objective：To study the relationship between apolipoprotein AI （ApoAI） and apolipoprotein B （ApoB） gene polymorphism and the syndrome of nontraumatic osteonecrosis of femoral head （NONFH）. Methods：By collecting 143 patients of NONFH and 92 normal control, restriction fragment length polymorphism-polymerase chain reaction （RFLP-PCR） technique was used to analyze the genotype of the ApoAI gene （-75bp in promoter and ＋83bp in intron） and ApoB gene （locus EcoRI, XbaI） and the VNTR polymorphism of ApoB gene was analyzed by PCR and agar gel electrophoresis. According to pattern differentiation of the Traditional Chinese Medicine of NONFH （the stagnation of phlegm and blood stasis in collateral branch of the large channel type, blockage of channels type, deficiency of liver-yin and kidney-yin type, analyze the frequency of genotype and allelomorph on the syndrome type statistically with SAS. Results： At -75bp in Promoter of ApoAI, the frequency of A/A genotype in blockage of channels type group （Group A） and deficiency of liver-yin and kidney-yin type group （Group B） was higher than that in control group （P0.05） and G/A genotype frequency in Group B was obviously lower than that in control group （P0.01）, the stagnation of phlegm and blood stasis in collateral branch of the large channel type group （Group C） and Group A. G/G genotype frequency in group B was lower than that in Group A （P0.05）. To compare with other groups, G allelomorph frequency in Group A was lower obviously. At promoter-intron of Apol AI, comparing with Group C, G/G-C/C, G/A-C/C genotype frequency in Group A and G/A-C/C genotype frequency was significantly lower while G/A-C/C genotype frequency in Group B was higher than Group A. In XbaI gene type of ApoB, X＋X-genotype frequency in Group A was significantly higher than that in control group （P0.05）, but X＋, X-allelomorph frequency was no evidently different （P0.05）. Comparing genotype frequency and allelomorph frequency in the ＋83bp loci of ApoAI gene and EcoRI loci, 3＇-VNTR of ApoB gene, there＇s no evident difference in each group （P0.05）. Conclusion：Locus -75bp and Locus promoter--intron polymorphism of ApoAI gene, XbaI loci of ApoB gene were associated with the syndrome type of NONFH. But no potential relationship was found between the syndrome type of NONFH and gene polymorphism of ＋83bp loci of ApoAI gene, EcoRI loci of ApoB gene or 3＇-VNTR of ApoB gene.