Wnt信号通路中Dkk1等在绝经后2型糖尿病女性患者中的表达与骨代谢关系的研究

发布时间：2018-01-29 05:47

本文关键词： 绝经后女性 2型糖尿病 Dkk1 β-catenin ELISA qRT-PCR 骨密度　出处：《石河子大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:通过检测绝经后女性2型糖尿病(type 2 diabetic mellium,T2DM)患者外周血白细胞Wnt信号通路中Dickkopf-1(Dkk1)基因及蛋白表达水平、β连环蛋白(β-catenin)基因表达水平,同时测定骨密度(bone mineral density,BMD)。探讨并分析相互关系,从而进一步探讨绝经后女性T2DM患者Wnt信号通路与骨代谢的关系。方法:1.收集2015年10月-2016年10月间新疆石河子地区社区及住院病人T2DM与糖耐量正常(Normal Glucose Tolerance,NGT)样本各50例,测定记录其一般资料,包括性别、年龄、身高、体重、收缩压、舒张压等一般临床基线资料,根据公式计算体重指数(Body Mass index,BMI)。EDTA抗凝管采空腹静脉血,全自动生化分析仪测定空腹血糖(Fasting Plasma Glucose,FPG)、血清胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)等生化指标,高压液相色谱法测定糖化血红蛋白(HbA1c);双能X线(Dual-energy X-ray absorptiometry,DEXA)法测定腰椎及股骨骨密度。2.采用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)测定血清中Dkk1蛋白表达水平。3.采用荧光定量逆转录聚合酶链式反应法(Quantitative Real-time PCR,qRT-PCR)检测Dkk1基因、β-catenin基因表达水平。4.使用SPSS 17.0软件包,计量资料以均数±标准差(sx±)表示,正态分布数据,两组均数间比较采用独立样本t检验。非正态分布数据,使用非参数检验。数据相关性分析采用多元线性回归分析。应用GraphPad Prism5对数据作图。以P0.05认为差异有统计学意义。结果:1.与绝经后女性NGT组相比,T2DM组年龄、SBP、DBP以及BMI均无统计学差异(P0.05),说明资料基线齐,具有可比性。2.与NGT组相比,T2DM组糖代谢指标:空腹血糖(Fasting Plasma Glucose,FPG)(5.47±0.73,9.26±2.38)、果糖胺(Froctosamine,FMN)(239.74±36.54,337.66±87.31)、HbA1c(6.23±0.92,8.70±1.76)(P0.01)水平增高;骨代谢指标:血钙(Ca),碱性磷酸酶(ALP)差异无统计学意义。磷(P)(1.11±0.12,1.24±0.16)水平较高(P0.01)、骨钙素(OC)(30.25±7.41,19.27±5.07)、25(OH)VitD3(35.37±6.83,22.00±5.26)(P0.01)水平较低,差异具有统计学意义。3.与NGT组相比,Fins水平(8.20±2.72,9.29±2.57)、HOMA-IR水平(2.00±0.70,3.75±1.14)较高;Fincp水平(0.48±0.10,0.30±0.09)降低;ISI、HOMA-β显著降低(P0.01);4.与NGT组相比,T2DM组BMD(股骨)密度显著降低。5.与NGT组相比,T2DM组Dkk1蛋白表达水平较高(209.37±32.35,232.97±39.31)(P0.05);外周血白细胞中Dkk1基因表达水平较高,Dkk1为NGT组的5.99倍。6.与NGT组相比,T2DM组外周血白细胞中β-catenin基因表达水平降低,T2DM组β-catenin基因表达水平为NGT组的0.46倍。7.Pearson相关性分析显示,在绝经后女性中,BMD(L2)与FMN(r=-0.204;p=0.042)呈负相关关系、BMD(L3)与25(OH)Vit D3(r=0.201;p=0.045)呈正相关关系、BMD(Femur)与FPG(r=-0.344;p=0.000)呈负相关关系,BMD(Femur)与Fincp(r=0.244;p=0.025)、HOMA-β(r=0.298;p=0.003)呈正相关关系;OC与FPG(r=-0.527;p=0.000)、FMN(r=-0.450;p=0.000)、HbA1c(r=-0.459;p=0.000)、HOMA-IR(r=-0.423;p=0.000)呈负相关关系,与Fincp(r=0.558;p=0.000)、ISI(r=0.376;p=0.000)、HOMA-β(r=0.419;p=0.000)呈正相关关系;25(OH)VitD3与FPG(r=-0.573;p=0.000)、FMN(r=-0.408;p=0.000)、HbA1c(r=-0.472;p=0.000)、HOMA-IR(r=-0.590;p=0.000)呈负相关关系,与Fincp(r=0.495;p=0.000)、ISI(r=0.312;p=0.000)、HOMA-β(r=0.391;p=0.000)呈正相关关系;P与Dkk1蛋白表达(r=0.301;p=0.013)呈正相关关系。8.以BMD(股骨)为应变量,以年龄、SBP、DBP、BMI、FPG、FMN、TG、TC、LDL-C、HDL-C等指标为自变量,进行多元线性逐步回归分析,在T2DM组中,Fincp进入回归方程。建立多元线性回归方程:y=1.377-0.339Fincp。结论:1.绝经后女性T2DM患者骨密度水平较低,FPG、FMN、Fincp、25(OH)VitD3均是其影响因素。2.Dkk1蛋白及基因的高表达导致β-catenin基因的低表达,参与绝经后女性T2DM的骨代谢异常。
[Abstract]:Objective : To detect the expression level of Dkk1 gene and protein in peripheral blood leukocyte Wnt signaling pathway in postmenopausal women with type 2 diabetic mellium ( T2DM ) , and to measure bone mineral density ( BMD ) . The relationship between Wnt signaling pathway and bone metabolism in postmenopausal women with T2DM was discussed and analyzed . Methods : 1 . 50 patients with T2DM and normal glucose tolerance ( NGT ) samples were collected from October 2015 to October 2016 in Shihezi district , Xinjiang , and 50 cases of normal Glucose Tolerance ( NGT ) samples were collected . Their general data , including sex , age , height , body weight , systolic pressure and diastolic blood pressure were measured . Body mass index ( BMI ) was calculated according to the formula . The levels of Dkk1 and 尾 - catenin in serum were determined by enzyme - linked immunosorbent assay ( ELISA ) . There was a negative correlation between OC and FPG ( r = - 0.527 ; p = 0.000 ) , FMN ( r = - 0.450 ; p = 0.000 ) , HbA1c ( r = - 0.459 ; p = 0.000 ) , ISI ( r = - 0.459 ; p = 0.000 ) ; Conclusion : 1 . The high expression of Dkk1 protein and gene leads to the low expression of 尾 - catenin gene , which is involved in the abnormal bone metabolism of women with T2DM after menopause .

【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1;R580

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