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维生素D3对子痫前期高风险患者外周血单核细胞Toll样受体4的影响
Effect of cholecalciferol supplementation on TLR4 expression on peripheral blood monocytes of pregnant women at risk for pre-eclampsia
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DOI:
作者:
汪洪友
Wang Hongyou
作者单位:
江苏省滨海县人民医院
Binhai County People’s Hospital,Jiangsu Province
关键词:
子痫前期;维生素D3;单核细胞;Toll样受体4
Pre-eclampsia; Monocytes; Toll-like receptor4; Vitamin D3
摘要:
目的:通过对子痫前期(PE)高风险患者补充维生素D3,检测外周血单核细胞Toll样受体4(TLR4)表达及其分泌促炎细胞因子,阐明维生素D3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例子痫前期高风险患者,随机双盲分为二组,从孕20~32周共持续12周,维生素D组每日服用2000 IU维生素D3,对照组服用安慰剂。孕32周肘前静脉抽取静脉外周血4 mL,流式细胞术(FCM)检测外周血单核细胞TLR4表达;分离外周血单核细胞经脂多糖(LPS)刺激18小时后,收集培养上清液,Luminex液相芯片检测肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6和IL-10浓度;并分析安慰剂组单核细胞TLR4阳性细胞比例与外周血清25-羟维生素D浓度的相关性。结果:与安慰剂组比较,Vitamin D组单核细胞TLR4阳性细胞频率和平均荧光强度均显著降低,差异均有统计学意义(P<0.05)。单核细胞与LPS共培养上清液,维生素D组TNF-α、IL-6浓度明显低于安慰剂组,IL-10浓度明显高于安慰剂组,差异均有统计学意义(P<0.05);安慰剂组单核细胞TLR4阳性频率与25-羟维生素D浓度呈负相关,相关系数为-0.532 (P= 0.002)。结论:PE高风险患者经补充维生素D3,可降低外周血单核细胞TLR4的表达,从而分泌促炎细胞因子IL-6和TNF-α减少,显著降低了PE的发病率。因此,通过补充维生素D3抑制单核细胞TLR4表达可能是预防子痫前期的新途经。
Objective: To clarify the mechanisms of cholecalciferol supplementation on preventing the occurrence of preeclampsia, the expression of Toll-like receptor (TLR4) and pro-inflammatory cytokines production in peripheral blood monocytes from pregnant women at risk for pre-eclampsia was investigated. 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. Subjects were randomly divided into 2 groups to receive 2000 IU vitamin D3 supplements (n=30) or receive placebo (n=30) from 20 to 32 weeks of gestation after all participants gave informed written consent. 4 mL of fresh venous blood was collected and the expression of TLR4 on monocytes was measured by flow cytometry (FCM). Moreover, monocytes were stimulated with LPS for 18 hours and the concentrations of IL-6, IL-10 and TNFα in supernatants were analyzed with Luminex platform. The correlation between the frequencies of TLR4+ monocytes and serum 25(OH)D levels was evaluated in placebo-controlled group. Results: After 12 weeks of cholecalciferol administration, the percentage of TLR4+ monocytes, as well as the mean fluorescence intension of TLR4, was markedly lower in vitamin D3 group than the placebo group (P<0.05). Stimulated by 50 ng/mL LPS for 18 hours, monocytes from vitamin D3 group produced less TNF-α, IL-6 and more IL-10 than the cells isolated from placebo group (P<0.05). In addition, a negative correlation was found between the percentage of TLR4+ monocytes and the serum levels of 25(OH)D in placebo group (r=-0.532, P= 0.0025). Conclusions: Our results showed that monocytes from pregnant women at risk for preeclampsia exhibited reduced TLR4 expression and produceed less pro-inflammatory cytokines upon LPS stimulation after 12 weeks vitamin D3 supplementation, and the incidence of pre-eclampsia was decreased. Thus, modulating TLR4 expression on monocytes by cholecalciferol supplementation may provide a new strategy to prevent pre-eclampsia.
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