人脐静脉内皮细胞的补体攻膜复合物C5b-9模型建立

Modeling of complement membrane attack complex C5b-9 in human umbilical vein endothelial cells

目的建立人脐静脉内皮细胞(HUVEC)的补体攻膜复合物C5b-9模型并确定其亚溶破剂量。方法通过体外培养HUVEC,在其表面组装攻膜复合物C5b-9,激光共聚焦检测其组装情况;增加补体浓度,通过激光共聚焦定性观察细胞上清液中乳酸脱氢酶(LDH)活性定量检测,确定其亚溶破剂量。结果激光共聚焦结果显示C5b-9成功组装于HUVEC表面;当C5b6的浓度达到1.6 mg·L-1时,LDH的分泌量骤增,激光共聚焦显示该剂量下部分细胞胞膜破裂。结论成功建立人脐静脉内皮细胞的补体攻膜复合物C5b-9模型,为研究动脉粥样硬化药物防治奠定基础。

Aim To establish complement membrane attack complex C5b-9 in human umbilical vein endothelial cells （HU- VEC） and determine its sublytie dose. Method HUVEC in vitro was cultured, and C5b-9 was established on its surface. Then the assembly result was observed by laser confoeal scan- ning microscope （LCSM）. To increase the concentration of com- plement, C5b-9 was established on HUVEC again. Through ob- serving the assembly condition by LCSM and detecting the con- centration of Lactate Dehydrogenase in cell supernatant, the sub- lytic dose of C5b-9 was gained. Results The LCSM results showed that C5b-9 had been successfully assembled in HUVECsurface. When the concentration of C5b6 came to 1.6 mg· L^-1, LDH＇s concentration surged, and LCSM showed that at this dose, part of the （,,ell membrane ruptured. Conclusion C5b-9 on HUVEC surface can be successfully established, which lays the foundation for the drug prevention and treatment of atherosclerosis in the future.