青蒿琥酯体外抑制手足口病合并脑炎患儿单个核细胞分泌血管内皮生长因子作用的实验研究

In vitro inhibitory effect of artesunate on the vascular endothelial growth factor secreted by mononuclear cells in hand-foot-and-mouth disease children complicated with encephalitis

目的探讨白细胞计数（WBC）、前白蛋白（PA）、超敏C反应蛋白（hs—CRP）、血管内皮生长因子（VEGF）在手足口病（HFMD）合并脑炎患儿中的变化规律，以及青蒿琥酯在体外对HFMD患者VEGF的抑制作用。方法采用Ficoll密度梯度离心法分离健康对照、无合并症的HFMD和HFMD合并脑炎患者单个核细胞，用不同浓度的青蒿琥酯（25、50、100mg／L）进行药物处理，采用ELISA双抗体夹心法检测各组细胞培养上清液中VEGF含量，同时检测3组患儿WBC、PA、hs—CRP水平。多个样本均数比较采用单因素方差分析，多个样本均数间的两两比较采用DunnettT3检验。两变量间相关性分析采用Spearman相关。结果无合并症的HFMD组、HFMD合并脑炎组和健康对照组间WBC、hs—CRP、PA、VEGF水平的差异均有统计学意义（F值分别为172．69、366．02、166．32和5941．89，均P〈0．01）。在3种不同浓度青蒿琥酯作用下，无合并症的HFMD组（F=194．265）、HFMD合并脑炎组（F=4750．69）单个核细胞分泌VEGF含量的差异均有统计学意义（均P〈0．01）。HFMD合并脑炎组、无合并症的HFMD组患儿单个核细胞分泌VEGF的含量与青蒿琥酯浓度均呈负相关（r值分别为-0．903、-0．969，均P〈0．01）。结论HFMD合并脑炎患儿WBC、hs—CRP、单个核细胞分泌VEGF含量均较无并发症HFMD患儿明显升高，PA明显下降。青蒿琥酯在体外能呈剂量依赖性地抑制手足口病患儿单个核细胞分泌VEGF。

Objective To investigate the dynamics of white blood cell count （WBC）, prealbumin （PA）, high sensitive C-reactive protein （hs-CRP）, and vascular endothelial growth factor （VEGF） in hand-foot-and-mouth disease （HFMD） complicated with or without encephalitis and in vitro inhibitory effects of artesunate on VEGF secretion of mononuclear cells from HFMD patients, Methods Peripheral blood mononuclear ceils from healthy control group, HFMD group and HFMD combined with encephalitis group were isolated by Fieoll density gradient centrifugation and treated with different concentrations of artesunate （25, 50, 100 mg/L）. The expression of VEGF in the supernatant was examined by ELISA double antibody sandwich method. The levels of WBC, PA and hs-CRP in the three groups were also detected. Multiple samples were compared by one-way analysis of variance. Multiple comparison was performed by Dunnett T3 test. Correlation of two variables was analyzed by Spearman correlation test. Results There were significant differences in the levels of WBC, hs-CRP, PA and VEGF between the HFMD combined with or without encephalitis group and health control＂group （F= 172. 69, 366. 02, 166.32 and 5 941. 89, respectively, all P^0. 01）. There were significant differences in the levels of VEGF secreted by mononuclear cells treated with three different concentrations of artesunate between the HFMD combined with or without encephalitis groups and health control group （F = 194. 265 and 4 750.69, respectively, both P^0.01）. Correlation analysis showed that the VEGF level secretion by mononuclear cells from HFMD patients with or without encephalitis were both negatively correlated with different concentrations of artesunate （r=--0. 903 and --0. 969, both P＇~0.01）. Conclusions Compared with HFMD without encephalitis, the secretion of VEGF by mononuclear cells, WBC and hs-CRP levels in HFMD complicated with encephalitis group all increase and PA decrease significantly. Artesunate can inhibit the secretion of VEGF by mononuclear cells of patients with hand-foot-and-mouth disease in a dose- dependent manner in vitro.