【摘要】： 目的:①探討漢族人群中多藥耐藥基因1(multidrug resistance gene 1,MDR1) C3435T位點多態(tài)性對外周血單個核細(xì)胞(peripheral blood mononuclear cells,PBMCs)中mRNA表達(dá)的影響。②通過研究苯巴比妥(phenobarbitone,PB)對PBMCs MDR1表達(dá)的影響,了解PBMCs MDR1的表達(dá)是否也能被誘導(dǎo),并研究了PBMCs中MDR1與孕烷X受體(pregnane X receptor,PXR)表達(dá)的關(guān)系,初步探討其意義。 方法:①以163名(男性98名,女性65名)無親緣關(guān)系的中國健康漢族兒童的外周血標(biāo)本為研究對象,采用PCR-RFLP技術(shù)檢測其MDR1基因C3435T位點的基因型,實時熒光定量PCR法檢測其PBMCs MDR1 mRNA表達(dá)水平。②將36名健康兒童的PBMCs分離后,將其分編為無培養(yǎng)組(不進(jìn)行培養(yǎng)和藥物處理)、自然培養(yǎng)組(只單純培養(yǎng)不進(jìn)行藥物處理)及PB培養(yǎng)組(加入終濃度為40μm/L苯巴比妥處理)。將自然培養(yǎng)組與PB培養(yǎng)組培養(yǎng)24h后,實時熒光定量PCR檢測這三組細(xì)胞的MDR1及PXR mRNA表達(dá)水平。③Hardy-Weinberg檢驗采用卡方檢驗。不同分組間的PBMCs MDR1或PXR表達(dá)水平的比較采用t檢驗或單因素方差分析,MDR1及PXR的表達(dá)水平相關(guān)性分析使用Spearman法。P0.05表示差異有統(tǒng)計學(xué)意義。 結(jié)果:①在163名健康中國漢族兒童中,63名為野生型純合子3435CC型,78名為雜合子CT型,22名為突變型純合子TT型,CC、CT與TT各基因型的發(fā)生頻率分別為38.7%、47.9%與13.5%;其C等位基因頻率為62.6%,T等位基因頻率為37.4%;C3435T多態(tài)性位點的基因型頻率分布符合Hardy-Weinberg平衡(P0.05);CC、CT和TT各基因型組間PBMCs MDR1 mRNA表達(dá)水平分別為(4.562±3.383)×10-3、(4.673±3.710)×10-3和(4.489±2.928)×10-3,差異無統(tǒng)計學(xué)意義(P0.05)。②未培養(yǎng)組、自發(fā)培養(yǎng)組和PB培養(yǎng)組MDR1 mRNA表達(dá)水平分別為(4.475±2.980)×10-3、(4.991±4.165)×10-3及(7.265±5.067)×10-3 , PXR mRNA在以上三組中的表達(dá)水平分別為(2.073±1.335)×10-3、(1.836±1.467)×10-3及(3.014±2.238)×10-3。PB培養(yǎng)組MDR1和PXR表達(dá)水平均高于未培養(yǎng)組和自發(fā)培養(yǎng)組,差異有統(tǒng)計學(xué)意義(P0.05),而未培養(yǎng)組和自發(fā)培養(yǎng)組之間的MDR1和PXR表達(dá)水平差異無統(tǒng)計學(xué)意義(P0.05);相關(guān)性分析提示,苯巴比妥處理后MDR1及PXR mRNA的表達(dá)水平呈正相關(guān)關(guān)系(r=0.517,P=0.0012)。 結(jié)論:①漢族人群中MDR1基因C3435T多態(tài)性對PBMCs mRNA表達(dá)并沒有顯著性影響,對于漢族兒童C3435T位點多態(tài)性不能作為推測MDR1表達(dá)水平的依據(jù)。②苯巴比妥可誘導(dǎo)PBMCs MDR1表達(dá),提示PBMCs MDR1表達(dá)也能被誘導(dǎo)。苯巴比妥誘導(dǎo)后PXR與MDR1的表達(dá)水平呈正相關(guān)關(guān)系,提示PXR可能參與了PBMCs MDR1基因的表達(dá)調(diào)控。
[Abstract]:Objective: 1 to investigate the effect of multidrug resistance gene 1 (multidrug resistance gene 1, MDR1) C3435T polymorphism in peripheral blood mononuclear cells (peripheral blood mononuclear cells,PBMCs) on the expression of mRNA in peripheral blood mononuclear cells (peripheral blood mononuclear cells,PBMCs) in Han population. (2) the expression of phenobarbital (phenobarbitone,) was studied by studying the expression of MDR1 in peripheral blood mononuclear cells (PBMCs). The effect of PB) on the expression of PBMCs MDR1 was investigated to see if the expression of PBMCs MDR1 could also be induced, and the relationship between the expression of MDR1 and progesterone X receptor (pregnane X receptor,PXR in PBMCs was studied, and its significance was discussed. Methods: 1 the peripheral blood samples of 163 unrelated Chinese healthy Han children (98 males and 65 females) were used to detect the genotype of C3435T locus of MDR1 gene by PCR-RFLP. The expression of PBMCs MDR1 mRNA was detected by real-time fluorescence quantitative PCR. (2) 36 healthy children were divided into non-culture group (no culture and drug treatment) after PBMCs was isolated and divided into non-culture group (no culture and drug treatment). Natural culture group (simple culture without drug treatment) and PB culture group (with final concentration of 40 渭 m / L phenobarbital). The expression levels of MDR1 and PXR mRNA were detected by real-time fluorescence quantitative PCR after culture in natural culture group and PB culture group for 24 hours. The 3Hardy-Weinberg test was performed by chi-square test. T test or single factor variance analysis were used to compare the expression levels of PBMCs MDR1 or PXR among different groups. Spearman method was used to analyze the correlation between the expression levels of MDR1 and PXR. P 0.05 showed that the difference was statistically significant. Results: 1among 163 healthy Chinese Han children, 63 were wild type homozygote 3435CC, 78 were heterozygote CT and 22 were mutant homozygote TT. The frequency of CC,CT and TT was 38.7%, respectively. 47.9% and 13.5%; The frequency of C allele and T allele were 62.6% and 37.4%, respectively. The genotype frequencies of C allele and T allele were in accordance with Hardy-Weinberg equilibrium (P0.05). The expression levels of PBMCs MDR1 mRNA in CC,CT and TT genotypic groups were (4.562 鹵3.383) 脳 10 鈮，

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