脂多糖对大鼠肺微血管内皮细胞血管性血友病因子血管性血友病因子裂解蛋白酶表达影响及维生素 D 的干预作用

摘要

Objective To observe the effects of lipopolysaccharide(LPS)on cultured rat pul⁃monary microvascular endothelial cells(PMVEC)von Willebrand factor(VWF), A disintegrin- like andmetalloproteinase with thrombospond in type-1 domain 13(ADAMTS13)expression, and the inter⁃vention effect of vitamin D(Vit D). Methods Primary cultured rat PMVEC were randomly assigned to 6 groups: NC group, LPS group(given 100 μg /mL LPS)and calcitriol(Cal)group(given 100 nmol/L Cal), LPS+Cal 1, 2, 3 groups(treated with 100 μg /mL LPS and 5 nmol/l, 20 nmol/L, 100 nmol/L Cal) were used to detect the expression of VWF and ADAMTS13 mRNA and protein in cell lysates. Results VWF, ADAMTS13 mRNA and protein expression showed no significant changes in LPS group and Cal group, VWF mRNA and protein expression levels in LPS group and LPS + Cal groups were significantly higher than those in NC group; VWF mRNA and protein expression levels in LPS + Cal groups of were significantly lower than those in LPS group; ADAMTS13 mRNA and protein expression levels in LPS group and LPS + Cal groups were significantly lower than those in NC group; ADAMTS13 mRNA and protein expression levels in LPS+Cal 3 group were significantly higher than those in the LPS group. Con-clusion Vit D in physiological condition does not change the expression of VWF and ADAMTS13 in PMVECs, while after being stimulated by LPS Vit D can prevent the increase of VWF expression and the reduction of ADAMTS13 caused by LPS. In the pathogenesis of sepsis or ALI, Vit D may play a role in the disease outcome by affecting the expression of VWF and ADAMTS13 in PMVEC.%目的：观察脂多糖（lipopolysaccharide，LPS）对体外培养大鼠肺微血管内皮细胞（pulmonary microvascular endothelial cells，PMVEC）内血管性血友病因子（von willebrand factor， VWF）、血管性血友病因子裂解蛋白酶（A disintegrin-like andmetalloproteinase with thrombospond in type-1 domain 13，ADAMTS13）表达的影响，以及维生素 D（vitamin D，Vit D）的干预作用。方法体外原代培养大鼠 PMVEC，随机分为6组：NC 组、LPS 组（给予100μg/mL LPS）、骨化三醇（Calcitriol，Cal）组（给予100 nmol/L Cal）及LPS+Cal 1、2、3组（分别给予100μg/mL LPS 和5 nmol/L、20 nmol/L、100 nmol/L Cal），分别检测细胞裂解液中VWF、ADAMTS13 mRNA和蛋白表达情况。结果 LPS 组和 Cal 组 VWF、ADAMTS13 mRNA 和蛋白表达均无显著变化，LPS 组和 LPS+ Cal 各组VWF mRNA、蛋白表达较NC组显著增高，LPS+ Cal各组VWF mRNA、蛋白表达较LPS组显著降低， LPS 组和 LPS+ Cal 各组 ADAMTS13 mRNA、蛋白表达较 NC 组显著降低，LPS+Cal 三组 ADAMTS13 mRNA、蛋白表达较 LPS 组显著增高。结论 Vit D 在生理状态下并不能改变 PMVEC 中 VWF 和ADAMTS13的表达，但在受到LPS刺激后可使已增高的VWF表达降低，已降低的ADAMTS13表达增加，在脓毒症或急性肺损伤（acute lung injury，ALI）发病过程中，补充 Vit D 可能通过对 PMVEC中VWF、ADAMTS13表达的影响对疾病的转归起作用。