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中华妇幼临床医学杂志(电子版) ›› 2018, Vol. 14 ›› Issue (04) : 398 -405. doi: 10.3877/cma.j.issn.1673-5250.2018.04.005

所属专题： 文献；

论著

妊娠期糖尿病患者白细胞NOS2及SFTPD基因表达与糖代谢异常的相关性分析
马小星1,†(), 张政梅1   
  1. 1. 716000　陕西，延安市人民医院产科
  • 收稿日期:2017-10-24 修回日期:2018-05-10 出版日期:2018-08-01
  • 通信作者: 马小星

Correlation analysis of NOS2 and SFTPD gene expression in leukocytes and abnormal glucose metabolism in pregnant women with gestational diabetes mellitus

Xiaoxing Ma1,†(), Zhengmei Zhang1   

  1. 1. Department of Obstetrics, Yan′an People′s Hospital, Yan′an 716000, Shaanxi Province, China
  • Received:2017-10-24 Revised:2018-05-10 Published:2018-08-01
  • Corresponding author: Xiaoxing Ma
  • About author:
    Corresponding author: Ma Xiaoxing, Email:
  • Supported by:
    Natural Science Research Project of Education Department of Shaanxi Provincial Government(2013JK0782)
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引用本文：

马小星, 张政梅. 妊娠期糖尿病患者白细胞NOS2及SFTPD基因表达与糖代谢异常的相关性分析[J]. 中华妇幼临床医学杂志(电子版), 2018, 14(04): 398-405.

Xiaoxing Ma, Zhengmei Zhang. Correlation analysis of NOS2 and SFTPD gene expression in leukocytes and abnormal glucose metabolism in pregnant women with gestational diabetes mellitus[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2018, 14(04): 398-405.

目的

探讨妊娠期糖尿病(GDM)患者白细胞NOS2及SFTPD基因与糖代谢异常的相关性。

方法

选择2014年1月至2016年12月，延安市人民医院妇产科收治的125例孕妇为研究对象。根据孕妇是否罹患GDM，将其分别纳入GDM组(n＝87)和非GDM组(n＝38)。采集2组受试者空腹肘静脉血，并分离白细胞，采用实时荧光定量PCR测定受试者白细胞NOS2和SFTPD mRNA相对表达水平。采集2组受试者孕前、分娩前体重，孕前人体质量指数(BMI)，血清三酰甘油(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、C反应蛋白(CRP)，糖化血红蛋白(HbA1c)，空腹血糖，口服葡萄糖耐量试验(OGTT) 1、2 h血糖，血浆胰岛素，稳态模型评估的胰岛素抵抗指数(HOMA-IR)，以及稳态模型评估的胰岛β细胞分泌功能指数(HOMA-β)。采用成组t检验，对2组孕妇上述指标进行比较。对于2组受试者白细胞NOS2和SFTPD mRNA相对表达水平与相关临床指标的相关性分析，采用Pearson相关性分析。采用Ingenuity路径分析(IPA)数据库构建与白细胞NOS2、SFTPD基因相关的分子网络，对其相关生物学功能进行分析。本研究符合2013年修订的《世界医学协会赫尔辛基宣言》，征得2组受试者知情同意，并签署临床研究知情同意书。2组受试者年龄、孕前BMI、孕期体重增加值等一般临床资料比较，差异均无统计学意义(P>0.05)。

结果

①GDM组受试者空腹及OGTT 1、2 h血糖值与HbA1c值分别为(89.9±18.6) mg/dL、(187.3±30.0) mg/dL、(163.3±22.3) mg/dL、(5.4±0.4)%，均分别显著高于非GDM组的(77.5±7.8) mg/dL、(152.3±36.1) mg/dL、(113.9±21.3) mg/dL、(5.2±0.4)%，并且差异均有统计学意义(t＝3.95、P<0.001, t＝5.63、P<0.001、t＝11.55、P<0.001，t＝2.57、P＝0.021)。②GDM组受试者白细胞NOS2和SFTPD mRNA相对表达水平分别为(0.08±0.05)、(0.16±0.07)，均显著高于非GDM组的(0.04±0.03)、(0.06±0.02)，并且差异均有统计学意义(t＝8.92、3.26，P<0.001)。③2组受试者白细胞NOS2和SFTPD mRNA相对表达水平与妊娠期血清CRP均呈弱正相关关系(r＝0.23、P＝0.022, r＝0.34、P<0.001)；SFTPD mRNA相对表达水平与空腹血糖、HOMA-IR均呈弱正相关关系(r＝0.19、P＝0.041, r＝0.21、P＝0.040)。白细胞NOS2与SFTPD mRNA相对表达水平呈强正相关关系(r＝0.77、P<0.001)。④在IPA数据库中检索到12种相关因素与白细胞NOS2基因过表达相关，而未检索到与白细胞SFTPD基因过表达相关因素。12种白细胞NOS2基因相关因素与免疫、炎症反应及血管生成相关，可分为4类：细胞/生长因子、趋化因子、酶、转录因子。

结论

GDM孕妇葡萄糖代谢受损，可能是其白细胞NOS2和SFTPD基因过表达的主要预测因子。NOS2和SFTPD基因表达异常与孕期葡萄糖代谢功能障碍和(或)炎症有关。本研究确定了12种与NOS2基因相关的调控免疫、炎症反应及血管生成的细胞因子、酶和转录因子。

关键词: 糖尿病，妊娠, 白细胞, NOS2基因, SFTPD基因, 糖代谢, 孕妇
Objective

To explore correlations between expressions of NOS2 and SFTPD gene in leukocytes and abnormal glucose metabolism changes in pregnant women with gestational diabetes mellitus (GDM).

Methods

From January 2014 to December 2016, a total of 125 pregnant women in the Department of Obstetrics and Gynecology, Yan′an People′s Hospital were selected as study subjects. According to whether they were GDM or not, they were divided into GDM group (n=87) and non-GDM group (n=38). Fasting cubital venous blood samples of subjects in two groups were collected and leukocytes were separated from the blood samples. The body weight before pregnancy and delivery, body mass index (BMI) before pregnancy; serum triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), C reaction protein (CRP); glycosylated hemoglobin A1c (HbA1c), fasting plasma glucose, oral glucose tolerance test (OGTT) 1 h and 2 h blood glucose; plasma insulin, homeostasis model assessment of insulin resistance (HOMA-IR) and homeostasis model assessment of β cell function index (HOMA-β) of two groups were collected and compared by independent-samples t test. The correlations between expressions of NOS2 and SFTPD gene in leukocytes of two groups and related clinical indicators were analyzed by Pearson correlation analysis method. The Ingenuity Pathway Analysis (IPA) database was used to construct molecular networks related to NOS2 and SFTPD gene, and their related biological functions were analyzed. This study met the requirements of the World Medical Association Declaration of Helsinki revised in 2013. Informed consent was obtained from every subject and every subject signed the informed consent form for clinical research. There were no statistical differences between two groups in the aspect of age, BMI before pregnancy and weight gain during pregnancy (P>0.05).

Results

①The values of fasting plasma glucose, OGTT 1 h and 2 h blood glucose, and value of HbA1c in GDM group were (89.9±18.6) mg/dL, (187.3±30.0) mg/dL, (163.3±22.3) mg/dL, (5.4±0.4)%, which were significantly higher than those in non-GDM group [(77.5±7.8) mg/dL, (152.3±36.1) mg/dL, (113.9±21.3) mg/dL, (5.2±0.4)%, respectively], and all the differences were statistically significant (t=3.95, P<0.001; t=5.63, P<0.001; t=11.55, P<0.001; t=2.57, P=0.021). ②The relative expression levels of NOS2 and SFTPD mRNA in leukocytes of GDM group were (0.08±0.05) and (0.16±0.07), respectively, which were significantly higher than those in non-GDM group [(0.04±0.03) and (0.06±0.02)], and both the differences were statistically significant (t=8.92, 3.26; P<0.001). ③The relative expression levels of NOS2 and SFTPD mRNA in leukocytes of the two groups showed weak positive correlations with serum CRP during pregnancy (r=0.23, P=0.022; r=0.34, P<0.001); and there were weak positive correlations between relative expression of SFTPD mRNA and value of fasting blood glucose, HOMA-IR (r=0.19, P=0.041; r=0.21, P=0.040). There was a strong positive correlation between the relative expression levels of NOS2 and SFTPD mRNA (r=0.77, P<0.001). ④In IPA database, a total of 12 factors related to NOS2 gene in leukocytes were retrieved, while no factor related to SFTPD gene in leukocytes was retrieved. Twelve NOS2 gene-related factors were associated with immune, inflammatory response and angiogenesis and can be divided into four categories: cells/growth factors, chemokines, enzymes, and transcription factors.

Conclusions

Impaired glucose metabolism in GDM patients may be the main predictor of the over-expression of NOS2 and SFTPD gene. In addition, abnormal expression of NOS2 and SFTPD gene are related to glucose metabolism dysfunction and (or) inflammation during pregnancy. This study identifies 12 cytokines, enzymes, and transcription factors that regulate the immune, inflammatory response, and angiogenesis associated with the NOS2 gene.

Key words: Diabetes, gestational, Leukocytes, NOS2 gene, SFTPD gene, Glucose metabolism, Pregnant women
表1 实时荧光定量PCR扩增引物序列
基因名称 引物序列(5′-3′) 扩增产物长度(bp)
NOS2 正向引物：CCCAAATACGAGTGGTTTCG 133
? 反向引物：TCTCTGTGCCCATGTACCA ?
SFTPD 正向引物：TGCTTTCCTGAGCATGACTG 164
? 反向引物：AAGCCCTGTCATTCCACTTG ?
GAPDH 正向引物：GGTGGTCTCCTCTGACTTCAACA 127
? 反向引物：GTTGCTGTAGCCAAATTCGTTGT ?
表2 2组孕妇一般临床资料及实验室检测指标比较(±s)
组别 例数 年龄(岁) 孕前BMI(kg/m2) 孕期体重增加值(kg) 脂质代谢指标
血清TG(mg/dL) 血清TC(mg/dL) 血清HDL-C(mg/dL) 血清LDL-C(mg/dL) HbA1c(%)
GDM组 87 29.9±4.6 25.0±6.5 10.6±8.2 245.1±79.6 265.5±52.8 71.6±21.6 152.5±56.1 5.4±0.4
非GDM组 38 28.6±4.6 24.2±4.5 9.9±5.4 224.1±63.2 259.7±41.8 79.2±15.2 145.0±63.4 5.2±0.4
t值 ? 1.45 0.69 0.48 1.44 0.60 1.96 0.66 2.57
P值 ? 0.151 0.449 0.559 0.242 0.628 0.101 0.625 0.021
组别 例数 空腹血糖(mg/dL) OGTT 1 h血糖(mg/dL) OGTT 2 h血糖(mg/dL) 血清CRP(mg/dL) 胰岛素抵抗和分泌指标
血浆胰岛素(μIU/mL) HOMA-IR HOMA-β
GDM组 87 89.9±18.6 187.3±30.0 163.3±22.3 4.4±3.9 9.0±10.7 2.3±2.9 142.2±195.9
非GDM组 38 77.5±7.8 152.3±36.1 113.9±21.3 5.2±4.9 8.1±9.4 1.6±1.8 242.5±315.7
t值 ? 3.95 5.63 11.55 0.97 0.45 1.37 1.16
P值 ? <0.001 <0.001 <0.001 0.448 0.645 0.164 0.110
表3 2组孕妇白细胞NOS2和SFTPD mRNA相对表达水平比较(±s)
组别 例数 NOS2 mRNA SFTPD mRNA
GDM组 87 0.08±0.05 0.16±0.07
非GDM组 38 0.04±0.03 0.06±0.02
t值 ? 8.92 3.26
P值 ? <0.001 0.001
图1 2组孕妇白细胞NOS2和SFTPD mRNA相对表达水平(图1A：NOS2 mRNA相对表达水平；图1B：SFTPD mRNA相对表达水平)
表4 2组孕妇白细胞NOS2和SFTPD mRNA相对表达水平与相关临床指标的相关性分析
临床指标 NOS2 mRNA SFTPD mRNA
r值 P值 r值 P值
年龄(岁) 0.16 0.073 0.02 0.812
孕前BMI(kg/m2) 0.07 0.453 －0.01 0.990
孕期体重增加值(kg) 0.06 0.529 0.13 0.160
血清TG(mg/dL) －0.14 0.274 －0.12 0.328
血清TC(mg/dL) －0.21 0.096 －0.12 0.322
血清HDL-C(mg/ dL) －0.24 0.067 －0.17 0.166
血清LDL-C(mg/dL) －0.10 0.445 －0.06 0.636
HbA1c(%) 0.12 0.208 0.13 0.156
空腹血糖(mg/dL) 0.17 0.070 0.19 0.041
OGTT 1 h血糖(mg/dL) 0.14 0.181 0.09 0.361
OGTT 2 h血糖(mg/dL) 0.08 0.393 0.07 0.450
血清CRP(mg/dL) 0.23 0.022 0.34 <0.001
血浆胰岛素(μIU/mL) 0.05 0.627 0.19 0.059
HOMA-IR 0.07 0.486 0.21 0.040
HOMA-β －0.08 0.442 －0.04 0.743
图2 采用Ingenuity路径分析数据库分析和构建的白细胞NOS2基因相关分子网络图
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