妊娠期运动干预对超重及肥胖孕妇Toll样受体4通路的下调作用

Exercises down regulate Toll-like receptor 4 signaling pathway in obese and overweight pregnant women

目的:探讨妊娠期运动干预对超重及肥胖孕妇Toll样受体4(Toll-like receptor 4,TLR4)通路的调控作用。方法:本研究基于本课题组2014年12月至2016年7月于北京大学第一医院开展的随机对照试验,选取其中择期剖宫产分娩,且除外合并妊娠并发症孕妇,最终纳入运动组12例、对照组11例。采用实时荧光定量聚合酶链反应、蛋白质印迹技术、流式细胞微珠法等检测方法比较2组孕妇外周血单个核细胞(peripheral blood mononuclear cell,PBMC)、腹直肌、网膜脂肪和皮下脂肪中 TLR4-髓样细胞分化因子(myeloid differentiation factor 8,MyD88)-核因子-κB(nuclear factor-κB,NF-κB)通路的表达情况,以及血浆中炎症因子的表达水平。采用独立样本 t检验、广义估计方程、 χ^(2)检验、Pearson相关分析等对数据进行统计学分析。 结果:(1)运动组孕妇中、晚孕期外周血浆中炎症因子TNF-α及IL-1β的表达呈现低于对照组的趋势,但差异均无统计学意义( P值均>0.05)。(2)运动组孕妇孕期PBMC中TLR4、MyD88和NF-κB的mRNA以及TLR4和NF-κB蛋白表达水平显著低于对照组(TLR4 mRNA:中孕期0.06±0.03与0.10±0.04,晚孕期0.05±0.02与0.11±0.05, χ^(2)=8.07;MyD88 mRNA:中孕期0.09±0.03与0.11±0.03,晚孕期0.10±0.04与0.17±0.06, χ^(2)=5.81;NF-κB mRNA:中孕期0.10±0.03与0.17±0.08,晚孕期0.08±0.03与0.20±0.08, χ^(2)=14.71;TLR4蛋白:中孕期1.7±0.5与1.9±0.8,孕晚期1.7±0.4与2.3±0.8, χ^(2)=5.83;NF-κB蛋白:中孕期1.0±0.4与1.5±0.4,晚孕期1.2±0.3与1.5±0.5, χ^(2)=4.73, P值均<0.05)。且随时间变化,运动组与对照组间PBMC中TLR4、MyD88和NF-κB的mRNA以及TLR4蛋白表达水平的差别逐渐增大。(3)运动组腹直肌、网膜脂肪组织中NF-κB(0.04±0.02与0.08±0.04, t=-3.72;0.25±0.05与0.63±0.21, t=-5.41;P值均<0.05)及皮下脂肪组织中TLR4和MyD88的mRNA水平明显低于对照组(0.12±0.03与0.30±0.10, t=-5.30;0.24±0.09与0.44±0.08, t=-5.38;P值均<0.05)。运动组网膜脂肪中MyD88和NF-κB及皮下脂肪中NF-κB蛋白水平明显低于对照组(1.1±0.5与2.0±0.8, t=-3.15;1.3±0.5与2.0±0.9, t=-2.23;1.2±0.5与1.9±0.8, t=-2.80;P值均<0.05)。(4)2组孕妇腹直肌中TLR4和NF-κB mRNA表达水平( r值分别为0.453和0.485)、网膜脂肪中NF-κB mRNA表达水平以及TLR4和MyD88蛋白水平( r值分别为0.539、0.437和0.527)与孕妇空腹血糖水平呈正相关( P值均<0.05)。 结论:妊娠期规律运动可以下调超重和肥胖孕妇TLR4-MyD88-NF-κB通路的表达和激活,且相关通路因子表达与空腹血糖水平具有一定相关性。

Objective To explore the effect of exercise intervention on regulation of Toll-like receptor 4(TLR4)signaling pathway in overweight and obese pregnant women.Methods The cohort was based on a randomized controlled trial(RCT)carried out by the same research group in Peking University First Hospital from December 2014 to July 2016.Overweight and obese patients who delivered by elective cesarean section without pregnancy complications were recruited,among which 12 cases in the exercise group and 11 cases in the control group were selected.Real-time polymerase chain reaction,Western Blot,and Luminex experiments were used to compare the expression of TLR4-myeloid differentiation factor 8(MyD88)-nuclear factor-κB(NF-κB)pathway in peripheral blood mononuclear cell(PBMC),rectus abdominis muscle,omental adipose,and subcutaneous adipose,as well as the levels of inflammatory factors(TNF-α,IL-1β,IL-10)in plasma between the two groups.Two independent samples t-test,generalized estimating equation,Chi-square test,and Pearson correlation analysis were adopted for statistical analysis.Results(1)The expression of inflammatory factors TNF-αand IL-1βin the exercise group showed a downward trend compared with the control in the second and third trimester,but none of the differences were statistically significant(all P>0.05).(2)The mRNA expression of TLR4,MyD88,and NF-κB and the protein expression of TLR4 and NF-κB in PBMC of the exercise group were significantly lower than those in the control group during pregnancy(TLR4 mRNA:0.06±0.03 vs 0.10±0.04 in the second trimester,0.05±0.02 vs 0.11±0.05 in the third trimester,χ^(2)=8.07;MyD88 mRNA:0.09±0.03 vs 0.11±0.03 in the second trimester,0.10±0.04 vs 0.17±0.06 in the third trimester,χ^(2)=5.81;NF-κB mRNA:0.10±0.03 vs 0.17±0.08 in the second trimester,0.08±0.03 vs 0.20±0.08 in the third trimester,χ^(2)=14.71;TLR4 protein:1.7±0.5 vs 1.9±0.8 in the second trimester,1.7±0.4 vs 2.3±0.8 in the third trimester,χ^(2)=5.83;NF-κB protein:1.0±0.4 vs 1.5±0.4 in the second trimester,1.2±0.3 vs 1.5±0.5 in the third trimester,χ^(2)=4.73;all P<0.05).Moreover,the differences in the mRNA expression of TLR4,MyD88,and NF-κB and TLR4 protein expression in PBMC between the two groups gradually increased.(3)NF-κB in rectus abdominis and omental adipose tissue(0.04±0.02 vs 0.08±0.04,t=-3.72;0.25±0.05 vs 0.63±0.21,t=-5.41;both P<0.05)and TLR4 and MyD88 in subcutaneous adipose tissue(0.12±0.03 vs 0.30±0.10,t=-5.30;0.24±0.09 vs 0.44±0.08,t=-5.38;both P<0.05)were observed a decreased mRNA level in the exercise group compared with the control group.The protein level of MyD88 and NF-κB in omental adipose tissue and NF-κB in subcutaneous adipose tissue in the exercise group were significantly lower than those in the control group(1.1±0.5 vs 2.0±0.8,t=-3.15;1.3±0.5 vs 2.0±0.9,t=-2.23;1.2±0.5 vs 1.9±0.8,t=-2.80,all P<0.05).(4)The expressions of TLR4 and NF-κB mRNA(r=0.453 and 0.485)in rectus abdominis muscle,NF-κB mRNA,TLR4 and MyD88 protein(r=0.539,0.437 and 0.527)in omental adipose in the two groups were positively correlated with the level of fasting blood glucose(P<0.05).Conclusions Regular exercise during pregnancy can down-regulate the expression and activation of the TLR4-MyD88-NFκB pathway in overweight and obese pregnant women.The expression of related factors along this pathway has a certain correlation with fasting blood glucose.