妊娠期糖尿病患者重要肠道菌群的特征分析及其与炎症因子的相关性研究

Characteristics of important intestinal flora and its correlation with inflammatory factors in gestational diabetes mellitus

目的观察GDM患者重要肠道菌群的变化特征,分析与炎症因子的相关性。方法选取GDM患者(GDM组)116例、妊娠期NGT女性(GNGT组)101名及非妊娠体检健康女性(NC组)80名。采用稀释平板计数法和快速细菌鉴定法对各组肠杆菌、肠球菌、双歧杆菌、乳杆菌、拟杆菌及梭杆菌进行定性、定量分析。采用ELISA和免疫荧光法测定TNF-α、IL-6、高敏C反应蛋白(hsC-RP)水平。Spearman相关分析肠道菌群与炎症因子的相关性。结果与NC组比较,GNGT组和GDM组BMI、SBP、DBP升高(P<0.01)。与GNGT组比较,GDM组SBP、DBP、FPG、HbA_1c、TC、TG、HDL-C均升高(P<0.05或P<0.01)。与NC组比较,GNGT组和GDM组肠杆菌[(7.14±0.28)vs(8.26±0.90)vs(9.54±1.08)logN/g]和拟杆菌[(7.99±1.46)vs(8.94±1.43)vs(9.45±1.68)logN/g]的菌落数增多,双歧杆菌[(10.47±0.18)vs(8.06±0.32)vs(7.35±0.49)logN/g]和乳杆菌[(7.52±1.36)vs(6.75±1.86)vs(5.31±0.68)logN/g]的菌落数降低(P<0.05或P<0.01)。GDM组肠杆菌和拟杆菌的菌落数较GNGT组增多,双歧杆菌和乳杆菌的菌落数降低,且肠球菌[(8.30±1.42)vs(6.81±0.57)、(7.12±1.80)logN/g]的菌落数高于NC组和GNGT组(P<0.05);GDM组TNF-α[(235.63±28.60)vs(126.77±12.05)、(123.43±14.92)ng/L]、IL-6[(147.73±24.26)vs(77.44±9.55)、(75.61±10.07)ng/L]及hsC-RP[(4.58±017)vs(1.21±0.20)、(1.05±0.19)mg/L]水平高于其他两组(P<0.01),而NC组和GNGT组比较,差异无统计学意义(P>0.05);Spearman相关分析发现,肠杆菌与TNF-α、IL-6及hsC-RP呈正相关(r=0.535、0.557、0.512,P<0.05或P<0.01);肠球菌与TNF-α和IL-6呈正相关(r=0.534、0.504,P<0.05);双歧杆菌(r=-0.681、-0.517)、乳杆菌(r=-0.425、-0.560)及拟杆菌(r=0.483、-0.432)与TNF-α和IL-6呈负相关(P<0.05或P<0.01);梭杆菌与TNF-α、IL-6及hsC-RP均无相关性(P>0.05)。结论妊娠期女性肠道益生菌减少,致病菌增加,GDM患者表现更为明显,推测肠道菌群可能在GDM的发生发展中发挥一定作用,且该作用可能与炎症因子水平变化有关。

Objective To investigate the changes of important intestinal flora in gestational diabetes mellitus,and analyze its correlation with inflammatory indicators. Methods A total of 297 subjects were enrolled in this study and divided into three groups： patients with GDM （GDM group, n=116）, pregnant women with normal glucose tolerance （GNGT group, n= 101）, and health women without pregnancy （NC group, n =80 ） . Enterobacteriaceae, Enterococcus, Bifidobacterium, Lactobacillus, Bacteroides and Fusobacterium were analyzed qualitatively and quantitatively by dilution plate count method and rapid bacterial identification method. The levels of TNF-a, IL-6 and hsC-RP were detected by ELISA. The correlation between intestinal flora and inflammatory indicators were analyzed by Spearmara Results The BMI,SBP and DBP were higher in GNGT group and GDM group than in NC group（P〈0. 01）. The SBP, DBP, FPG, H bA1 c, TC, TG and HDL-C were higher in GDM group than in GNGT group（P〈0. 05 or P〈0. 01）. The clump count of Enterobacteriaceae[（9.54 ± 1.08） vs （8. 26 ± 0.90） vs （7.14 ± 0. 28） logN/g] and Bacteroides [（9.45 ± 1.68） vs （8. 94 ± 1.43） vs （7.99± 1.46） logN/g] increased, and the Bifidobacterium[（7.35±0. 49） vs （8. 06±0. 32） vs （10.47±0.18）logN/g] and Lactobacillus[（5.31± 0. 68） vs （6.75±1.86） vs （7.52±1.36）logN/g] decreased in GDM and GNGT group than in NC group, （P〈 0. 05 or P〈0.01）. The Clump count of Enterobacteriaceae and Bacteroides were higher, and Bifidobacterium and Lactobacillus were lower in GDM group than in GNGT group, and the clump count of Enterocoeeus[-（8. 30±1.42） vs （6.81zk0. 57）, （7.12±1.80）logN/g] were higher in GDM group than in NC and GNGT groups （P 〈 0. 05）. The expression level of TNF-a [- （235.63 ± 28.60） vs （126.77 ± 12.05） ,（123.43±14.92）ng/L], IL-6[（147.73±24.26） vs （77.44±9.55）, （75.61±10.07） ng/L] and hsC-RP[ （4. 58 ± O17） vs （ 1.21 ±0. 20）, （ 1.05 ± 0. 19 ） mg/L] were higher in GDM group than in the other two groups （P〈0. 01）, and there were no statistical significances between NC and GNGT groups（P 〈0. 05）. Spearman correlation analysis showed that Enterobacteriaceae was positively correlation with TNF-a,IL-6 and hsC-RP （r= 0. 535,0. 557,0. 512, P〈0. 05 or P〈0. 01）. Enterococcus was positively correlation with TNF-a and IL-6 （r= 0. 534,0. 504, P〈0] 05）. Bifidobacterium（r= -0. 681, -0. 517）, Lactobaeillus（r= 0. 425, - 0. 560） and Bacteroides （r=- 0. 483, - 0. 432） were negatively correlated with TNF-a and IL-6（P〈0. 05 or P〈0. 01） .Fusobacterium was not correlated with TNF-a, IL-6 or hsC- RP （P〉0. 05）. Conclusion The probiotics decreases and pathogenic bacterium increases in pregnant women, especially in patients with GDM. Intestinal flora may play an important role in the occurrence and development of GDM, which is associated with the changes of inflammatory factors.