【作者】 黄澄；

【导师】 杨天伦；

【作者基本信息】 中南大学 ， 内科学， 2011， 博士

【摘要】 目的：研究湖南长株潭地区汉族人群中IRS-1Gly972Arg基因多态性与原发性高血压、胰岛素抵抗与内皮功能受损相互之间作用的关系。研究方法：选取湖南省长株潭地区510名经确诊的原发性高血压和胰岛素抵抗患者及520名健康对照者。其中单纯高血压患者184例,单纯胰岛素抵抗患者178例,高血压合并胰岛素抵抗患者145例,均为汉族人群。健康对照组选自于高血压或胰岛素抵抗患者同一地区的无血缘关系的群体,无高血压、糖尿病病史及家族史(计520例,其中男性271例,女性249例)。测量对象一般指标(如BMI、腰臀比、血压、血糖、血脂、肝肾功能等),并进行口服葡萄糖耐量试验(OGTT)评估受试者的胰岛素抵抗情况,使用HP Sonos5500彩色超声系统检测受试者肱动脉介导的血管舒张水平(FMD)。应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术,对受试者IRS-1Gly972Arg基因多态性进行基因分型。结果：IRS-1Gly972Arg基因多态性符合Hardy-Weinberg平衡定律。IRS-1基因Gly972Arg多态性的突变率(即G/A基因型频率)：原发性高血压患者中发现9例突变携带者(9/329),占2.7%,血压正常受试者中发现10例突变携带者(10/701),占1.4%；胰岛素抵抗患者中发现7例突变携带者(10/326),占2.1%,正常糖耐量受试者中发现12名突变携带者(12/704),占1.7%。尽管高血压组该基因突变率略高于血压正常组,胰岛素抵抗组略高于糖耐量正常组,但差异没有统计学意义。Logistic回归分析显示,经基因分型、胰岛素抵抗指数、高血压分级、吸烟、饮酒、年龄、性别、高血压家族史、血脂水平调整后,只有原发性高血压分级水平才是内皮功能受损的独立危险因素IRS-1Gly972Arg基因多态性、胰岛素抵抗以及两者之间的交互作用与内皮功能受损无明显相关性。结论：高血压水平分级是内皮功能受损的独立危险因素,胰岛素抵抗、IRS-1Gly972Arg基因多态性,以及两者之间的交互作用与内皮功能受损无明显相关性。目的：初步探讨IRS-1G972R变异致人群血管内皮功能受损的机制。方法：应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术,对受试者IRS-1Gly972Arg基因多态性进行基因分型。使用HP Sonos5500彩色超声系统检测受试者肱动脉介导的血管舒张水平(FMD)。ELISA法测定血清ET-1, eNos, IL-8和ICAM-1水平。外周血单个核细胞CXCR2mRNA表达水平通过Real-time PCR测定。数据进行多因素方差分析。结果：在1030例受试者中,G972R IRS-1等位基因突变频率为11.1%。IRS-1G972R基因多态性与内皮功能受损之间无明显的统计学差异。内皮功能受损与高血压及血浆ET-1/eNOS水平明显相关。然而,在高血压受试者中,G972R IRS-1突变者其血浆ET-1/eNOS水平显著高于Gly/Gly纯合子携带者。在胰岛素抵抗的受试对象中,与Gly/Gly纯合子携带者相比,G972R IRS-1突变者血清IL-8和ICAM-1水平和单核细胞CXCR2mRNA表达水平显著升高。结论：本研究显示,内皮功能受损的主效应因素是高血压,而不是IRS-1G972R多态性。然而,在高血压的受试者中,G972R IRS-1突变进一步增加了血清ET-1/eNOS水平, ET-1/eNOS水平与内皮功能状态密切相关；同时,在胰岛素抵抗的受试者中,G972R IRS-1突变促进了炎症反应,而炎症反应是内皮功能受损的另一个重要因素。总之,本研究结果显示,在内皮功能受损的进展中,IRS-1G972R突变与高血压之间存在交互作用；在炎症反应中,IRS-1G972R突变与胰岛素抵抗之间存在交互作用。目的：探讨IRS-1Gly972Arg基因多态性对于高血压或胰岛素抵抗受试者采用卡托普利和罗格列酮治疗后相关指标变化值的影响。方法：随机选取HBP,IR,HBP+IR和正常组各20例,每组按G/A基因型不同各分配10例。合并高血压组40名受试对象口服卡托普利,疗程一个月,每日25mg,比较治疗前后ET-1/eNOS表达水平变化；血浆合并胰岛素抵抗者口服罗格列酮,疗程2个月,每日4mg,比较治疗前后PBMC CXCR2mRNA表达水平变化。结果：服用卡托普利1月后各小组的ET-1/eNOS表达水平较未服药前降低,治疗前后ET-1/eNOS变化的绝对值水平在A等位基因受试人群中高于G等位基因的受试者。但对卡托普利降低其外周血浆ET-1/eNOs表达水平的效果采用协方差分析并未得出统计学意义。协方差分析还提示,是否合并胰岛素抵抗以及等位基因类型对高血压受试者卡托普利服用后FMD升高幅度影响不显著。服用罗格列酮2个月后各小组PBMC CXCR2mRNA表达水平较未服药前均下降,但协方差分析亦未得出罗格列酮对PBMC CXCR2mRNA表达水平影响的统计学意义；此外,是否合并高血压以及等位基因类型对胰岛素抵抗受试者服用罗格列酮后PBMC CXCR2mRNA表达水平降低幅度的影响并不显著。结论：在湖南省长株潭地区的汉族人群中,服用卡托普利能有效降低高血压受试者的ET-1/eNOs表达水平,并有效改善其内皮功能；是否合并胰岛素抵抗以及IRS-1Gly972Arg基因分型并不会影响ET-1/eNOs表达水平的下降程度。服用罗格列酮后胰岛素抵抗受试者的PBMC CXCR2mRNA表达水平显著下降,但受试者是否合并高血压以及IRS-1Gly972Arg等位基因分型对其下降幅度的影响无统计学意义。更多还原

【Abstract】 Objective:To investigate the interrelationship between IRS-1gene Gly972Arg polymorphism and primary hypertension,insulin resistance and endothelial dysfunction in Han people from Changsha,Zhuzhou and Xiangtan district.HuNan province.Methods:184hypertensive patients,178insulin-resistant patients,145hypertensive and insulin-resistant patients and520healthy controll people were recruited from Changsha,Zhuzhou and Xiangtan district.HuNan province. BMI, WHR, BP,BS; blood fat, hepatic and renal function were measured,insulin resistant state were assessed by OGTT. FMD were determined through HP Sonos5500color ultrasound system, IRS-1gene Gly972Arg polymorphism were detected by PCR-RFLP.Results:IRS-1gene Gly972Arg polymorphism coincide with Hardy-Weinberg equilibrium law. G/A allelic frequency:9heterogeneic carriers were detected from329hypertensive patients(2.7%),10were detected from701normotensive patients(1.4%);7were found in326insulin-resistant patients(2.1%),12were found in704subjects without insulin resistance(1.7%).Statistical difference can neither be found between hypertensive group and normotensive group nor between insulin-reisistant group and subjects without insulin resistance. Logistic regression analysis demonstrate that only hypertensive degree serves as independent risk factor of endothelial dysfunction (OR=1.983,95%CI1.215-2.573, P=0.028). No statistical relationship had been found between endothelial dysfunction and IRS-1Gly972Arg polymorphism,insulin resistance,nor the interaction between IRS-1Gly972Arg polymorphism&insulin resistance.Conclusion:Hypertensive degree serves as independent risk factor of endothelial dysfunction. No statistical relationship had been found between endothelial dysfunction and IRS-1Gly972Arg polymorphism,insulin resistance,nor the interaction between IRS-1Gly972Arg polymorphism&insulin resistance. Objective:To investigate a potential interrelationship between HBP、IRS-1G972R polymorphism、insulin-resistance and even FMD in vivo.Methods:a eight cohorts of10Gly/Gly carriers without HBP or IR,10Gly/Gly carriers with HBP but without insulin resistance,10Gly/Gly carriers with insulin resistance but without HBP、10Gly/Gly carriers with HBP and insulin resistance,10heterozygous carriers without HBP or insulin resistance,10heterozygous carriers with HBP but without insulin resistance,10heterozygous carriers with insulin resistance but without HBP、10heterozygous carriers with HBP and insulin resistance were enrolled respectively. The study population were Han people from Hunan Province,China.IRS-1polymorphism was detected by PCR.Endothelium-dependent vasodilution in terms of FMD were assessed by noninvasive color ultrasound system. ET-1、eNos and IL-8, ICAM-1were determined by enzyme linked immunosorbent assay,while the expression of mononuclearcell CXCR2mRNA were measured by Real-time PCR.Results:Allelic mutation frequency of G972R IRS-1allele were11.1%.No statistical difference were found between IRS-1G972R polymorphism and endothelial dysfunction. Statistical interrelationship lies between endothelial dysfunction and hypertension or plasma ET-1/eNOS level.However,the plasma ET-1/eNOS level of G972R IRS-1variants is statistically higher than that of Gly/Gly carriers,as detected in hypertensive subjects. Plasma IL-8and ICAM-1,as well as PBMC CXCR2mRNA elevated in G972R IRS-1variants compared with Gly/Gly carriers,as detected in insulin-resistant subjects.Conclusion:Through inflammation as well as interaction with HBP, IRS-1G972R polymorphism contribute to impaired FMD, indirectly, and hence cardiovascular disease. OBJECTIVE:To investigate the effects of IRS-1Gly972Arg polymorphism on hypertensive or insulin-resistant subjects treated by captopril or rosiglitazone.Methods:20hypertensive patients,20insulin-resistant patients,20hypertensive and insulin-resistant group and20healthy controll subjects were enrolled randomly as4groups.Each group was subdivided according to G/A genotype.40hypertensive subjects had25mg captopril daily for oral use for1month, the alleosis of plasma of ET-1/eNOS between pre-and post-captopril treatment were analyzed bystatistics.40insulin-resistant subjects had4mg rosiglitazone daily for oral use for2month, the change of PBMC CXCR2mRNA between pre-and post-rosiglitazone treatmentwere also compared through covariance analysis.Results:ET-1/eNOS level decreased after1month of captopril treatment,but no statistical difference was made in the effects of captopril on plasmaET-1/eNOs level,as demonstrated by covariance analysis.Also, covariance analysis showed that insulin resistance and allele had no statistical influence on effects of captopril on FMD elevation. PBMC CXCR2mRNA level declined after2months of rosiglitazone adminstration,but no statistical difference was made in the effect of rosiglitazone on PBMC CXCR2mRNA expression; In addition, covariance analysis revealled that hypertension and allele had no statistical contribution on effects of rosiglitazone on PBMC CXCR2mRNA degression.Conclusion:Captopril adminstration could decrease plasma ET-1/eNOs level of hypertensive subjects and improve endothelial function;Insulin resistance and IRS-1Gly972Arg polymorphism had no influence on the effect of captopril on depressing plasmaET-1/eNOs level. Rosiglitazone adminstration may downregulate PBMC CXCR2mRNA in insulin-resistant subjects,but neither hypertension nor IRS-1Gly972Arg allele showed statistical contribution on the effects of rosiglitazone on PBMC CXCR2mRNA degression.更多还原