维生素D_3对先兆子痫高风险孕妇外周血单核细胞Toll样受体4的影响  

Effect of cholecalciferol supplementation on Toll-like receptor 4 expression in peripheral blood monocytes from pregnant women at risk for preeclampsia

作　　者：汪洪友[1] 吴凤会[1] 周建波[1] 王晓春[1] 钱雷[2] WANG Hongyou;WU Fenghui;ZHOU Jianbo;WANG Xiaochun;QIAN Lei(Department of Obstetrics and Gynecology, People ＇ s Hospital of Binhai County, Yancheng 224500, Jiangsu , Chin)

机构地区：[1]江苏省盐城市滨海县人民医院妇产科,江苏盐城224500 [2]江苏省盐城市滨海县人民医院检验科,江苏盐城224500

出　　处：《上海医学》2018年第3期155-160,共6页Shanghai Medical Journal

摘　　要：目的通过对先兆子痫(PE)高风险孕妇补充维生素D_3,检测其外周血单核细胞Toll样受体4(TLR4)表达及其分泌促炎性细胞因子的情况,阐明维生素D_3在预防PE发生过程中的作用。方法根据子宫动脉异常多普勒波形,选取60例PE高风险孕妇,随机双盲分入维生素D_3组和安慰剂组两组,每组30例,自孕20~32周起,维生素D组每日服用2 000U维生素D_3,安慰剂组服用安慰剂,均持续服用12周。12周后采集两组孕妇肘前静脉外周血4mL,采用电化学发光免疫法检测外周血血清25-羟维生素D[25(OH)D]质量浓度,采用流式细胞术(FCM)检测外周血单核细胞TLR4表达(以平均荧光强度和细胞阳性率表示),并分析安慰剂组单核细胞TLR4阳性率与外周血血清25(OH)D质量浓度的相关性。再将两组孕妇各分为2个亚组(每组15例),分离外周血单核细胞分别经0或50ng/mL脂多糖(LPS)刺激18h后,收集培养上清液,通过液相芯片分析系统检测TNF-α、IL-6和IL-10的质量浓度。计算两组孕妇的PE发病率。结果维生素D_3补充12周后,维生素D_3组25(OH)D的质量浓度为(34.91±4.36)μg/L,显著高于安慰剂组同时间的(16.09±4.51)μg/L(P<0.05);维生素D_3组的单核细胞TLR4阳性率和平均荧光强度分别为23.6%(17.4%,38.6%)和26.7(20.5,38.9),均显著低于安慰剂组的38.7%(21.4%,60.5%)和37.4(22.3,56.8),差异具有统计学意义(P值均<0.05)。安慰剂组单核细胞TLR4阳性率与PE孕妇外周血血清25(OH)D质量浓度呈负相关(r=-0.532,P=0.002 5)。经LPS刺激培养18h后,维生素D_30ng/mL亚组和维生素D_350ng/mL亚组的上清液TNF-α、IL-6、IL-8质量浓度分别显著低于安慰剂同剂量亚组,IL-10质量浓度显著高于安慰剂同剂量亚组(P值均<0.05)。维生素D_3组外周血TNF-α、IL-8和IL-6的质量浓度分别显著低于安慰剂组(P值均<0.05),IL-10的质量浓度显著高于安慰剂组(P<0.05)。维生素D_3组的PE发病率为20.0%(6/30),显著低于安慰剂组的43.3%(13/3Objective To detect the expression of Toll-like receptor 4 （TLR4） and proinflammatory cytokine production in peripheral blood monocytes from pregnant women at risk for preeclampsia, and to clarify the mechanisms of cholecalciferol supplementation in preventing preeclampsia. Methods Randomized double-blind placebo-controlled clinical trial was performed among 60 pregnant women at risk for pre-eclampsia according to abnormal uterine artery Doppler waveform. The subjects randomly received 2 000 U vitamin D3 （n = 30, vitamin D3 group） or placebo （n = 30, placebo group） from 20 to 32 weeks of gestation. Venous blood （4 mL） was collected 12 weeks after administration of medicine, and the expression of TLR4 （expressed by mean fluorescence intensity and positive rate of cells） in monocytes was measured by flow cytometry （FCM）. The mass concentration of serum25（OH）D was determined by electrochemiluminescence immunoassay. The correlation between TLR4 positive rate in monocytes and serum 25（OH）D levels was evaluated in placebo group. Moreover, the subjects were divided in two subgroups （n = 15）. Then monocytes were stimulated with lipopolysaccheride （LPS, 0 or 50 ng/mL） for 18 h, and the concentrations of interleukin （IL）-6, IL-10 and tumor necrosis factor （TNF）-α in supernatants were analyzed with Luminex platform. The incidence of preeclampsia was recorded. Results After 12-week administration of medicine, the mass concentration of serum 25 （OH）D in vitamin D3 group was significantly higher than that in placebo group （E34.91 ± 4. 361 μg/L vs. [16.09 ± 4.51]μg/L, P〈0.05）. The TLR4 positive rate in monocytes and mean fluorescence intensity in vitamin D3 group were 23.6% （17.4%, 38.6%） and 26.7 （20.5, 38.9） respectively, which were significantly lower than those in placebo group （38.7 % [21.4 %, 60.5 % 1, 37.4 E22.3, 56.8%, both P〈0.05）. A negative correlation was found between the TLR4 positive rate in monocytes and the serum level of 25（OH）D i

关 键 词：先兆子痫 胆钙化醇 单核细胞 TOLL样受体4 

分 类 号：R714.244[医药卫生—妇产科学]