【作者】 张宁；

【导师】 叶元华；

【作者基本信息】 青岛大学 ， 妇产科学， 2007， 硕士

【摘要】 目的检测浆细胞膜糖蛋白(PC-1)及胰岛素受体酪氨酸激酶(IRTK)在妊娠期糖尿病(GDM)孕妇、正常孕妇及非孕对照妇女骨骼肌中的表达及活性，探讨二者与妊娠期糖尿病发病的关系。方法采用Western blot的方法测定了20例GDM孕妇(GDM组)、20例糖耐量正常孕妇(正常妊娠组)及12例糖耐量正常非孕妇女(对照组)骨骼肌组织PC-1、胰岛素受体的表达水平；采用免疫沉淀及ELISSA的方法分别测定胰岛素刺激前后骨骼肌组织胰岛素受体酪氨酸磷酸化程度及IRTK活性；采用葡萄糖氧化酶法及放射免疫法测定空腹血葡萄糖(FPG)及空腹血胰岛素(FINS)水平，计算胰岛素抵抗指数(HOMA-IR)。结果1)GDM组FPG、FINS、HOMA-IR(5.61±0.78mmol／L、15.12±5.31 mU／L、1.21±0.52)均明显高于正常妊娠组(4.43±0.46 mmol／L、10.56±3.07 mU／L、0.80±0.31)(P＜0.01)，正常妊娠组FINS、HOMA-IR明显高于对照组(7.56±2.31 mU／L、0.47±0.26)(P＜0.01)。2)GDM组PC-1表达水平(1.22±0.02)明显高于正常妊娠组(0.71±0.03)(P＜0.01)，正常妊娠组明显高于对照组(0.43±0.02)(P＜0.01)。3)GDM组胰岛素受体含量、胰岛素受体基础酪氨酸磷酸化程度及其酪氨酸激酶基础活性(0.59±0.03、0.14±0.03、2.33±0.19)与正常妊娠组(0.61±0.05、0.15±0.02、2.36±0.26)，正常妊娠组与对照组(0.63±0.06、0.16±0.04、2.39±0.35)比较，差异均无显著性(P＞0.05)。4)胰岛素刺激后的胰岛素受体酪氨酸磷酸化程度及IRTK活性比较，GDM组(0.17±0.04、2.86±0.47)明显低于正常妊娠组(0.24±0.02、3.84±0.53)(P＜0.01)，正常妊娠组明显低于对照组(0.31±0.03、4.87±0.56)(P＜0.01)。5)正常妊娠组、GDM组PC-1表达水平与HOMA-IR呈明显正相关(r=0.611、0.734，P＜0.01)，与胰岛素刺激后胰岛素受体酪氨酸激酶活性成负相关(r=-0.652、-0.723，P＜0.01)。结论1)妊娠期糖尿病患者骨骼肌组织中存在PC-1的高表达。2)骨骼肌组织PC-1高表达可能是妊娠期糖尿病患者胰岛素抵抗的分子机制之一。3)PC-1可能通过抑制胰岛素受体酪氨酸磷酸化及IRTK活性发挥作用。更多还原

【Abstract】 Objective To investigate the relationship between plasma cell membrane glycoprotein-l(PC-1) in skeletal muscle from women with GDM and the etiology of gestational diabetes mellitus.Methods The expression of PC-1 and insulin receptor in skeletal muscle were determined by western blot from patients with GDM(GDM group, n=20),normal pregnant women (normal pregnancy group,n=20)and nonpregnant women(control group, n=12), the expression of insulin receptor tyrosine phosphorylation and the activity of insulin receptor tyrosine kinase (IRTK) before and after insulin stimulation in skeletal muscle of the three groups were determined by immunoprecipitation and ELISSA.Fasting plasma glucose (FPG) and fasting insulin (FINS) were measured by oxidized assay and immunoradioassay. Insulin resistant index(HOMA-IR)was calculated according to FPG and FINS.Results1) The levels of FPG, FINS, HOMA-IR in GDM group(5.61±0.78 mmol/L、15.12±5.31 mU/L、1.21±0.52) were significantly higher than those in normal pregnancy group (4.43±0.46 mmol/L、10.56±3.07 mU/L、0.80±0.31)(P＜0.01).The levels of FINS and HOMA-IR in normal pregnancy group were significantly higher than those in control group (7.56±2.31 mU/L、0.47±0.26) (P＜0.01).2) PC-1 content in GDM group (1.22±0.02) was higher than that in normal pregnancy group (0.71±0.03) ,PC-1 content in normal pregnancy group was higher than that in control group (0.43±0.02) (P＜0.01).3) No significant differences were evident in insulin receptor content , basal insulin receptor tyrosine phosphorylation ,basal insulin receptor tyrosine kinase activity in GDM group (0.59±0.03、0.14±0.03、2.33±0.19), normal pregnancy group (0.61±0.05、0.15±0.02、2.36±0.26) and control group (0.63±0.06、0.16±0.04、2.39±0.35) (P＞0.05). 4) After insulin stimulation, insulin receptor tyrosine phosphorylation and insulin receptor tyrosine kinase activity increased in all subjects but were lower in GDM group (0.17±0.04、2.86±0.47) compared with normal pregnancy group (0.24±0.02, 3.84±0.53) and control group (0.31±0.03, 4.87±0.56) (P＜0.01).5) PC-1 content was positively correlated with HOMA-IR(r=0.611、0.734, P＜0.01) and negtively correlated with IRTK activity (r=-0.652、-0.723, P＜0.01) in normal pregnancy group and GDM group.Conclusions1) PC-1 content in skeletal muscle of GDM overexpresses.2) Overexpressed PC-1 in skeletal muscle may be one of the cellular mechanism for insulin resistance of GDM.3) PC-1 may produce the effect by inhibiting insulin receptor tyrosine phosphorylation and the activity of insulin receptor tyrosine kinase.更多还原