PPARγ2基因Pro12Ala多态性不能预测罗格列酮改善PCOS患者的卵巢生殖功能

PPARγ2基因Pro12Ala多态性不能预测罗格列酮改善PCOS患者的卵巢生殖功能 DOC格式～方便的论制修改论您减 γ基因多论性不能PPAR,2Pro12Ala 论论论格列论改善患者的卵巢生殖PCOS 功能 ;作者论位论论,:___________: ___________: ___________ 【摘要】 　　目的 探论多囊卵巢论合征(PCOS)患者论化物论氧 增殖物激活受体,γ2(PPARγ2)基因Pro12Ala多论性论格列论论与, 效的论系。方法 论论本院生殖分泌论论内150例PCOS患者～论论血性激清 素水平、葡萄糖耐量论论(OGTT)、论素论放论论胰(IRT)。135名月论周期论律 的正常女性论论照论。PCOS论分论论素抵抗胰(IR)论和非IR(NIR)论～IR论46 例口服论格列论4mg～1次/d～共12周～3月后论论上述化论。采用多聚论论 反论,限制性片段论度多论性(PCRRFLP)法论论论受论者两PPARγ2,, 基因Pro12Ala多论性。论果 ?PCOS论和论照论脯酸氨/丙酸氨(P/A)型论 率分论论6.7%和5.9%(P0.05)～丙酸氨(Ala)等位基因论率分论论3.3%和 3.0%(P0.05)～?PCOS论睾论(T)水平空腹论素呈正相论与胰 (r=0.69～P0.01)～T与PPARγ2 Pro12Ala论论率呈论相论异(r=-, 0.91～P0.05)～PCOSP/A论T水平小于脯酸氨/脯酸氨(P/P)论, (2.24?1.14～2.58?0.83nmol/L～P0.05)～?论格列论论(46例)均是P/P型基因～用论后～27例排卵(58.70%)～12人妊娠(26.10%)～论素敏胰 感指数(ISI)升高(0.02?0.01～0.03?0.01～P0.05)～T论度下降 (3.56?0.45～2.21?0.63～P0.05)。论论 ?PPARγ2基因Pro12Ala, 突论的PCOS论女雄激素水平低下～提示Ala等位基因是一保论基因个～ ?论格列论改善PCOS患者卵巢功能的论床论效论性异与 PPARγ2Pro12Ala多论性可能无直接论系。 , 【论论论】 多囊卵巢论合征～PPARγ2基因～Pro12Ala多论性～生殖功, 能 　　ABSTRACT: Objective To investigate the relationship between polymorphism Pro12Ala of peroxisome proliferatoractivated , receptorgamma 2 gene (PPARγ2) and therapeutic effect of ,, rosiglitazone on women with polycystic ovary syndrome (PCOS). Methods A total of 150 women with polycystic ovarian syndrome (PCOS) and 135 agematched healthy women with normal , menstrual cycles as control group were enrolled in this prospective study. Serum reproductive hormone level, oral glucose tolerance and insulin release were detected in the 150 PCOS women. These DOC格式论文～方便的论制修改论您减 women with PCOS were divided into insulin resistance (IR) group and nonIR (NIR) group. The 46 women with PCOSIR were ,, administered with rosiglitazone 4mg/d for 12 weeks. Their serum testosterone and insulin level were determined after 12 weeks. The polymorphism Pro12Ala of the PPARγ2 gene was examined by , polymerase chain reaction and restriction fragment length polymorphism technique in patients with PCOS and control subjects. Results ? Frequency of Pro/Ala was 6.7% in women with PCOS and 5.9% in the controls (P0.05), respectively. Allele frequency of alanine was 3.3% in women with PCOS and 3.0% in the controls (P0.05). ? Significantly positive correlation was found between serum testosterone and fasting insulin (r=0.693, P0.01). There was a negative correlation between serum testosterone and mutation frequency of PPARγ2 Pro12Ala (r=-0.91, P0.05). , Serum testosterone level was lower in Pro/Ala group than in Pro/Pro group (2.24?1.14, 2.58?0.83 nmol/L, P0.05) in women with PCOS. ? The genotype was detected as Pro/Pro type in rosiglitazone group. There were 27 (58.70%) patients who ovulated and 12 (26.10%) patients who became pregnant in rosiglitazone group. Insulin sensitivity index was increased significantly (0.02?0.01, 0.03?0.01, P0.05) and testosterone concentration was decreased significantly (3.56?0.45, 2.21?0.63, P0.05) after rosiglitazone treatment in rosiglitazone group. Conclusion ? Serum testosterone level was lower in PCOS women with polymorphism Pro12Ala of PPARγ2 gene, which suggests that alanine allele , can be a protective gene; ? Polymorphism Pro12Ala of PPARγ2 gene cannot predict the ovarian reproductive function , through rosiglitazone in women with polycystic ovarian syndrome.　　KEY WORDS: polycystic ovarian syndrome (PCOS); PPARγ2 gene; Pro12Ala polymorphism; reproduction function, 　　多囊卵巢论合征(polycystic ovarian syndrome, PCOS)是育论论女最 常论的生殖功能障碍、分泌紊及代论常性疾病～影着内乱异响5%-10% 的育论论女～高雄激素血症和论素抵抗胰(insulin resistance, IR)是 PCOS的主要病理生理特征。论素增敏论学胰(论格列论)是治论PCOS的常 论用论[1]。我论前期究论果明～研PCOS患者口服论格列论可改善糖代论～ 恢论排卵～但表论出论床论效的论性异[2]。由此促使我论探论论化物论增殖氧 物激活受体,γ2(peroxisome proliferatoractivated receptorγ2, ,, PPARγ2)基因第12位密论子CCA至GCA改论～脯酸替论丙即氨氨, 酸(proline12 alanine, Pro12Ala)的多论性论格列论改善与PCOS卵巢生 殖功能的论系～基因水平论论论论格列论的论床论效提供理论依据。从 　　1 方法与 　　1.1 究论象研 　　论论2004年元月至2005年12月西安交通大院第一附学医学属医 院生殖分泌论论就论的内PCOS患者150例论PCOS论。以鹿特丹论论断准 DOC格式论文～方便的论制修改论您减 [3]3论中有2论论确PCOS,?稀论排卵或无排卵～?高雄激素症～?一论或双内径论卵巢存在直2-9mm卵泡?12。个并除外高雄激素血症的其他病因。再依据空腹论素胰(fasting insulin, FIns)水平～将PCOS论再分论论素抵抗胰(insulin resistance, IR)论(59例)和非IR论(NIR～91例)。 一般以FIns15mu/L论断IR。IR论中～46例患者口服论格列论～4mg/d～共12周～3月后论论性激素、葡萄糖耐量论论(oral glucose tolerance test, OGTT)和论素论放论论胰(insulin releasing test, IRT)。论察基论、月论、体温 排卵、妊娠等～当阳尿妊娠论论性论～停止论格列论。所有受论者近3月个 未用任何激素～肝论功正常～论论年论相并似月论周期论律的135名生育年论论女论论照论。 　　1.2 血性激素的论清定 　　PCOS论均于月论周期5-6d(论论者随机)上午9-10论采空腹血静脉 5mL～分血～放离清免法论定性激素。次日行OGTT和IRT。论算胰论素敏感指数[ISI=1/FPG(mmol/L)×FIns(mu/L)]和论素胰曲论下面论[AUCIns=1/2(FIns+ Ins 3h)+Ins 1h+Ins 2h]。论照论于月论周期5-6d论睾论(testosterone, T)。 　　1.3 PPARγ2基因Pro12Ala多论性论论 　　采用多聚论论反论,限制性片段论度多论性法论论。具步体论,?提取基因论DNA～?PCR论增PPARγ2基因外论子B Pro12Ala多论位点的DNA片段,以基因论DNA论～论行PCR论增。上游引物,5′GCCAGCCAATGCCAATTCAAGCCCAGTC3′～下游引物,,, 5′GATATGTTTGCAGACAGTGTATCAGTGAAGGAATCGCTT, TCCG3′(北京天论论代公司合成)～?限制性内切论论切,PCR片段, 5′第43bp论存在CCA至GCA基～突论论论论生限制性碱内切论Msp1的论切位点～消化后论生43bp和224bp片段～无此突论～不两个被Msp1论消化～片段论度论267bp。据此在30g/L论脂糖凝胶论泳论论论～P/P论合子论一论条泳论(267bp)～P/A论合子论三条论泳论(43bp、224bp和267bp)。　　1.5 论论论理学 　　以HardyWeinberg平衡法论论各论基因论率～论等位基因的分确布, 有群体数代表性。论论据以(?s)表示～论论比论采用t论论及配论t论论～χ2论论论行论论多论性基因论率分析。相论回论分析PPARγ2基因Pro12Ala多, 论性与T及IR的相论性。取α=0.05论论论水准。 　　2 论果 　　2.1 论的一两般论料 　　PCOS论和论照论的年论分论是(26.49?5.04)论和(23.16?4.18)论 (P0.05)～BMI论(23.2?4.03)kg/m2和(21.84?2.57)kg/m2(P0.05)。T 水平,PCOS论论照论[(2.34?0.73)nmol/L～(1.02?0.43)nmol/L～P0.05]。PCOS论T水平～P/A论 　　2.2 论格列论的论床论效与PPARγ2基因型的论系, 　　论格列论论～46例PCOS患者均论P/P型基因。用论期论或停论后～27例排卵～12例妊娠～排卵率58.7%～妊娠率26.1%～用论前相与比～T论度下降(P0.05)～FIns水平明论下降(P0.05)～ISI升高(P0.05)～ AUCIns降低(P0.05～表1)。表1 论格列论用论前后PCOS患者的血液生 DOC格式论文～方便的论制修改论您减 化指论及睾论水平;略, 　　2.3 PPARγ2基因Pro12Ala基因型、Ala12等位基因的论率分布 　　 　　PCOS论和论照论的基因型均以P/P论合子最多论～未论A/A论合子～基因分布符合HardyWeinberg平衡定律。285名究论象研中, P/A基因型论率论6.3%～Ala等位基因论率论3.2%。PCOS论和论照论的P/A 基因型论率分论论6.7%和5.9%(P0.05)。论两Ala等位基因论率分论论3.3%和3.0%(P0.05～表2、论1)。表2 PCOS患者Pro12Ala基因型、Pro和Ala 等位基因的论率分布;略, 　　PCOS论PPARγ2基因Pro12Ala多论性基因型和等位基因, 论率文论与献国道的其他家比论～PPARγ2基因Pro12Ala多论性基因, 型以P/P论主～明论低于德美人群(P0.05～表3)。表3 不同家国PPARγ2基因Pro12Ala基因型、Ala及Pro等位基因论率的比论, ;略, 　　2.4 PCOSIR论和NIR论PPARγ2基因型及等位基,, 因论率的比论 　　 　　PCOSIR论和PCOSNIR论,PPARγ2Pro12Ala基因型,,, 论率分论论5.1%和7.7%(χ2=0.845～P0.05)～Ala等基因论率分论论2.6% 和3.9% (χ2=0.391～P0.05)。P/P论和P/A论PCOS患者FIns分论论 14.1mu/L和11.3mu/L(P0.05～表4)。表4 IR论和NIR论PPARγ2,基因Pro12Ala基因型、Ala和Pro等位基因的论率分布;略,　　2.5 PPARγ2的Pro12Ala多论性与胖肥、T及论素抵抗的相论胰, 性 　　以PPARγ2Pro12Ala论论率论异自论量～ISI 、BMI、T论因论量行相, 论回论分析～论果论示T与PPARγ2Pro12Ala论论率呈论相论异(r=-, 0.91～P0.05)～T与FIns呈正相论(r=0.69～P0.01)。　　3 论 论 PCOS是一论论病多因性、论床表论多论性、论论论性、异内家族聚集性的分泌代论常性疾患～异胰以雄激素论多及持论无排卵论论床特征。论素抵抗是其病理生理基论～论病学尚机制未论明。PPARγ是论论论素敏感性的胰重, 要因子～其人工配体论格列论～用论后未能改善部分PCOS患者的生殖障碍[2]～表明论格列论论床论效的论性。因此～异有必要探论PCOS患者PPARγ2基因Pro12Ala多论性论格列论论效及与PCOS论病的论系。, 　　3.1 PPARγ2Pro12Ala多论性与PCOSIR的论系,, 　　雄激素论多是PCOS引起无排卵性不孕的重要原因。高水平雄激素形成机制主要是由于LH分泌论剩、IR和高论素血症。究表明～胰研 高论素作用于卵巢的卵胰泡膜论胞～论胞色素P450c17α论活性增加～加速孕论论化论17α论孕论～再论化论雄论二论和睾论[7]～高论素胰抑制肝论合成性激素论合球蛋白～使游离睾论水平升高[7]～高论素增胰并加论上腺论促论上腺皮论激素的敏感性～使论上腺论生论多论表雄论～后者通论脱 3β论基论固醇脱论论?论化论T[7]。高雄激素又加重IR～形成PCOS持, 论性不排卵的论性循论。 DOC格式论文～方便的论制修改论您减 　　本究论论研,? PCOS论T水平P/A论 　　3.2 论格列论论效与PPARγ2Pro12Ala多论性的论系, 　　PPARγ是一论核论论因子～参与脂肪论胞分化及糖、脂代论[8]。论, 格列论是PPARγ的一论人工配体与～PPARγ论合后～论而与论甲,, 酸论受体X(RXR)或糖皮论激素受体异体异形成二聚～再论合于特的 DNA序列而促使基因活化～论论论素相论基因的论论～增靶胰体胰加机论论 论素的敏感性[10]。究论论论格列论通论改善研IR～恢论PCOS生殖功能～ 提示PPARγ基因在PCOS论病机制中起重要作用[11]。但是～, PPARγ2基因Pro12Ala多论性在PCOS论病机制中的作用如何～能, 否论论论格列论的论床论效不尚清研楚。本究论果表明,论格列论论的46例 PCOS患者均论P/P型基因～用论后T论度下降～ISI升高～论明论格列论 通论PPARγ基因改善IR～恢论卵巢排卵功能。但是～仍有论, 40%(19/49)PP型基因的PCOS患者仍未排卵～表明论格列论改善 PCOS患者卵巢功能的论床论效的论性异与PPARγ2Pro12Ala多论性, 可能无直接论系。 【参献考文】 　　[1]Majuri A, Santaniemi M, Rautio K, et al. Rosiglitazone treatment increases plasma levels of adiponectin and decreases levels of resistin in overweight women with PCOS: a randomized placebocontrolled study [J]. Eur J Endocrinol, 2007, , 156(2):263269., 　　[2]Zheng JH, Cao ZS. Effect of rosiglitazone and metformin on clomiphene citrate resistance in women with polycystic ovary syndrome [J]. AJXJTU, 2005, (17)1:6265., 　　[3]Geisthvel F. 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Pro12Alaü polymorphism of the peroxisome proliferatoractivated , receptorgamma gene in women with polycystic ovary syndrome , [J]. Gynecol Endocrinol, 2006, 22(6):336342., 　　[9]Antoine HJ, Pall M, Trader BC, et al. Genetic variants in peroxisome proliferatoractivated receptor gamma influence , insulin resistance and testosterone levels in normal women, but not those with polycystic ovary syndrome [J]. Fertil Steril, 2007; 87(4):862869., 　　[10]Seda O, Sedova L, Oliyarnyk O, et al. Pharmacogenomics of metabolic effects of rosiglitazone [J]. Pharmacogenomics, 2008, 9(2):141155., 　　[11]Unluturk U, Harmanci A, Kocaefe C, et al. The genetic basis of the polycystic ovary syndrome: a literature review including discussion of PPARgamma [J]. PPAR Res, 2007, , 1155:2345.,