心型脂肪酸结合蛋白对脓毒症血管内皮细胞损伤的影响

The effect of heart-type cytoplasmic fatty acid-binding proteinon vascular endothelial cell injury in sepsis

目的:探讨心型脂肪酸结合蛋白(H-FABP)对脓毒症血管内皮细胞损伤的影响。方法:体外培养人脐静脉内皮细胞(HUVECs),分别转染H-FABP小干扰RNA或过表达载体,构建下调或上调H-FABP的HUVECs,然后经100 ng/mL脂多糖(LPS)诱导12 h后,流式细胞术检测细胞凋亡,蛋白印迹法检测细胞中cleaved-caspase-9和cleaved-caspase-3蛋白表达,试剂盒检测细胞中SOD活性和MDA含量及细胞培养上清液中LDH、TNF-α和IL-1β水平。结果:LPS促进HUVECs中H-FABP蛋白表达(P<0.05),下调H-FABP降低LPS诱导的HUVECs凋亡率、cleaved-caspase-9和cleaved-caspase-3蛋白表达及LDH、MDA、TNF-α和IL-1β表达(P<0.05),提高SOD活性(P<0.05);上调H-FABP对LPS诱导的HUVECs凋亡、氧化应激及炎症因子表达的影响与下调H-FABP相反。结论:H-FABP可促进脓毒症血管内皮细胞损伤,其可能是治疗脓毒症的分子靶点。

Objective: To investigate the effect of heart-type cytoplasmic fatty acid-binding protein(H-FABP) on vascular endothelial cell damage in sepsis. Methods: Human umbilical vein endothelial cells(HUVECs) were cultured in vitro. H-FABP small interfering RNA or over expression vector was transfected into HUVECs to down-regulated or up-regulated H-FABP. Then lipopolysaccharide(LPS) was added to induced sepsis for 12 h.Cell apoptosis was detected by flow cytometry. The protein expression levels of cleaved-caspase-9 and cleavedcaspase-3 were detected by western blotting. The SOD activity and MDA content in cells and the levels of LDH,TNF-α and IL-1β in cell culture supernatant were detected. Results: LPS promoted the expression of H-FABP protein in HUVECs(P<0.05). Down-regulation of H-FABP decreased the apoptosis rate of HUVECs induced by LPS, the protein expression levels of cleaved-caspase-9 and cleaved-caspase-3, the levels of LDH, MDA, TNF-αand IL-1β(P<0.05), while increased SOD activity(P<0.05). The effects of up-regulating H-FABP on the apoptosis, oxidative stress and inflammatory factor levels in LPS-induced HUVECs were opposite to that of down-regulating H-FABP. Conclusion: H-FABP can promote the injury of vascular endothelial cells in sepsis, and it may be a molecular target for the treatment of sepsis.