CAJ | 학술논문

目的：研究β-胡萝卜素（β-C）对高糖诱导的内皮细胞损伤的保护作用，并初步探讨其作用机制。方法在血管内皮细胞长至80％以上融合时，将其分为5组：正常对照组、高糖损伤组及低、中和高剂量β-C组，每组6个复孔。正常对照组加入10％葡萄糖注射液5．5 mmol·L-1，高糖损伤组加入10％葡萄糖注射液33 mmol·L-1，低、中和高剂量β-C组分别加入β-C 10、20、40μmol·L-1＋10％葡萄糖注射液33 mmol·L-1。各组放置5％CO2培养箱中培养48 h后，收集细胞或培养液进行实验。使用酶联免疫检测仪、采用 MTT比色法检测5组血管内皮细胞存活率，采用流式细胞仪检测血管内皮细胞凋亡率、血管内皮细胞中活性氧（ROS）水平，使用全自动生化仪、采用乳酸→丙酮酸连续监测法检测5组血管内皮细胞培养液中乳酸脱氢酶（LDH）活性，使用全自动生化仪、采用硫代巴比妥酸比色定量法检测5组血管内皮细胞培养液中丙二醛（MDA）的水平；先参照 NO检测试剂盒的使用说明书进行实验，后采用酶联免疫检测仪检测5组血管内皮细胞培养液中 NO水平。结果与正常对照组比较，高糖损伤组的血管内皮细胞存活率、血管内皮细胞培养液中 NO水平均明显降低，血管内皮细胞培养液中 LDH 活性、MDA水平，血管内皮细胞凋亡率、血管内皮细胞中 ROS水平均明显升高（均P＜0．01）；与高糖损伤组比较，中、高剂量β-C组的血管内皮细胞存活率、血管内皮细胞培养液中 NO水平均明显升高，血管内皮细胞培养液中 LDH 活性、MDA水平及低、中和高剂量β-C组的血管内皮细胞凋亡率、血管内皮细胞中ROS水平均明显降低（P＜0．05或P＜0．01）。结论β-胡萝卜素对高糖诱导的脐静脉内皮细胞的损伤具有保护作用，其机制可能与增加 NO水平和促进 ROS的降解有关。
목적：연구β-호라복소（β-C）대고당유도적내피세포손상적보호작용，병초보탐토기작용궤제。방법재혈관내피세포장지80％이상융합시，장기분위5조：정상대조조、고당손상조급저、중화고제량β-C조，매조6개복공。정상대조조가입10％포도당주사액5．5 mmol·L-1，고당손상조가입10％포도당주사액33 mmol·L-1，저、중화고제량β-C조분별가입β-C 10、20、40μmol·L-1＋10％포도당주사액33 mmol·L-1。각조방치5％CO2배양상중배양48 h후，수집세포혹배양액진행실험。사용매련면역검측의、채용 MTT비색법검측5조혈관내피세포존활솔，채용류식세포의검측혈관내피세포조망솔、혈관내피세포중활성양（ROS）수평，사용전자동생화의、채용유산→병동산련속감측법검측5조혈관내피세포배양액중유산탈경매（LDH）활성，사용전자동생화의、채용류대파비타산비색정량법검측5조혈관내피세포배양액중병이철（MDA）적수평；선삼조 NO검측시제합적사용설명서진행실험，후채용매련면역검측의검측5조혈관내피세포배양액중 NO수평。결과여정상대조조비교，고당손상조적혈관내피세포존활솔、혈관내피세포배양액중 NO수평균명현강저，혈관내피세포배양액중 LDH 활성、MDA수평，혈관내피세포조망솔、혈관내피세포중 ROS수평균명현승고（균P＜0．01）；여고당손상조비교，중、고제량β-C조적혈관내피세포존활솔、혈관내피세포배양액중 NO수평균명현승고，혈관내피세포배양액중 LDH 활성、MDA수평급저、중화고제량β-C조적혈관내피세포조망솔、혈관내피세포중ROS수평균명현강저（P＜0．05혹P＜0．01）。결론β-호라복소대고당유도적제정맥내피세포적손상구유보호작용，기궤제가능여증가 NO수평화촉진 ROS적강해유관。
Objective To study the effects ofβ-carotene(β-C)on high glucose-induced endothe-lial cell damage and its mechanism of action.Methods When cell fusion reached more than 80%, vascular endothelial cells were treated with 5.5 mmol · L-1 10% glucose injection (control group),33 mmol·L-1 (high glucose group),10μmol·L-1β-C+ 33 mmol·L-1 glucose injection (low doseβ-C group),20μmol·L-1β-C + 33 mmol·L-1 glucose injection(medium doseβ-C group),or 40μmol·L-1β-C+ 33 mmol·L-1 glucose injection(high doseβ-C group).Each group included six wells.After culture for 48 hours in an incubator filled with 5%CO2 ,cells or culture medium were collected.Cell viability was assayed by MTT colorimetry using microplate reader. Cell apoptosis and intracellular reactive oxygen species(ROS)generation were detected by flow cytometry.The activity of lactic dehydrogenase in medium was examined by continuous monito-ring of the conversion lactic acid to pyruvic acid using automatic biochemical analyzer.Malondial-dehyde(MDA)production in medium was determined by TBARS assay using automatic biochemi-cal analyzer.Nitric oxide(NO)content in medium was measured by Griess Reagent assay usingautomatic biochemical analyzer.Results Compared with control group,cell viability and NO con-tent in medium significantly decreased whereas cell apoptosis,intracellular ROS generation and levels of MDA and LDH in medium markedly increased in high glucose group(P<0.01).Com-pared with high glucose group,cell viability and NO content in medium significantly increased whereas levels of MDA and LDH in medium markedly decreased in medium and high doseβ-C groups(P<0.05 or P<0.01).In addition,cell apoptosis and intracellular ROS generation obvi-ously decreased in low,medium and high doseβ-C groups,compared with high glucose group(P<0.05 or P<0.01).Conclusion The treatment withβ-carotene exerts protective roles in high glu-cose-induced umbilical vein endothelial cell inj ury through increasing NO production and promo-ting ROS degradation.