羟乙基淀粉130/0.4对内毒素诱导的大鼠血浆炎症因子表达和单个核细胞核转录因子活性的影响

Effects of hydroxyethyl starch 130/0.4 on serum inflammatory cytokines and transcription factor activation in blood monocytes in endotoxemic rats

目的观察羟乙基淀粉130/0.4(HES130/0.4)对内毒素腹腔注射大鼠炎症因子表达的影响,并通过其对单个核细胞核因子-κB(NF-κB)和激活蛋白-1(AP-1)活性的影响探讨相关分子机制。方法36只雄性Sprague-Dawley大鼠随机分为6组(n=6),对照组静脉输注生理盐水30ml/kg,LPS组腹腔注射脂多糖(LPS)5mg/kg后静脉输注生理盐水30ml/kg,LPS+HES组按照HES130/0.4的剂量不同分为3个亚组,分别腹腔注射LPS5mg/kg,接着分别静脉输注HES130/0.47.5、15、30ml/kg,HES组静脉输注HES130/0.430ml/kg。在腹腔注射LPS3h后以ELISA法检测血浆肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)、干扰素-γ(IFN-γ)浓度,用凝胶电迁移法检测NF-κB和AP-1的水平。结果腹腔注射LPS后大鼠血浆TNF-α、IL-10和IFN-γ的浓度较对照组明显升高,且NF-κB和AP-1的活性明显增加(P<0.05);HES130/0.4静脉输注可降低血浆TNF-α和IFN-γ的浓度以及NF-κB和AP-1的活性(P<0.05),而增加IL-10的浓度(P<0.05),其中15ml/kg剂量组对TNF-α、IFN-γ和NF-κB的作用最强(P<0.01),而7.5ml/kg剂量组对IL-10和AP-1的作用最强(P<0.01)。结论HES130/0.4可降低内毒素注射后促炎因子的表达,增强抗炎因子的表达,可能与其抑制转录因子NF-κB和AP-1的活性有关。

Objective To study the effects of hydroxyethyl starch （ HES 130/0.4） on serum inflammatory cytokines and the activities of nuclear factor KB （ NF - KB） and activator protein - 1 （ AP - 1 ） in blood monocytes in endotoxemic rats challenged with lipopolysaccharide （LPS）. Methods Male Sprague - Dawley rats were randomly divided into 6 groups with 6 animals in each group： （ 1 ） saline control; （2） LPS alone; （3,4 and 5） LPS plus HES （7.5, 15 and 30 ml/kg, respectively） ; （6） HES alone （30 ml/kg）. The rats were anesthetized with sodium pentobarbital. The tail vein was cannulated for continuous infusion. Animals were sacrificed 3 h after LPS. Serum tumour necrosis factor - α （ TNF - α） , interleukin - 10 （ IL - 10） , and interferon - γ （ IFN - γ） were measured by ELISA ; NF -κB and AP - 1 activities were measured by electrophoretic mobility shift assay. Results Three hours after LPS administration, serum TNF -α, IL - 10 and IFN - γ concentrations and NF - κB and AP - 1 activities were significantly increased compared with that in the saline control group. HES 7.5, 15, and 30 ml/kg significantly reduced the LPS - induced increments of TNF - α and IFN - γ levels and NF - κB and AP - 1 activities （ P 〈 0.05 ） , while further levated IL - 10 level （ P 〈 0.05 ）. HES 130/0.4 15 ml/kg caused the greatest reduction in TNF - α, IFN - γ and NF - KB （ P 〈 0.01 ）, while HES 130/0.4 7.5 ml/kg was most potent on regulating IL - 10 and AP - 1 （ P 〈 0.01 ）. Conclusion HES 130/0.4 can reduce serum pro - inflammatory cytokines and increase anti - inflammatory cytokine, probably by modulating the activation of NF - κB and AP - 1 in blood monocytes in endotoxemic rats.