南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2010年
2期
321-325
,共5页
熊莺%王广发%张俊艳%吴少瑜%徐伟%张嘉杰%吴曙光%饶进军
熊鶯%王廣髮%張俊豔%吳少瑜%徐偉%張嘉傑%吳曙光%饒進軍
웅앵%왕엄발%장준염%오소유%서위%장가걸%오서광%요진군
柚皮苷%人脐静脉内皮细胞%单核细胞%粘附分子%活性氧%转录因子%糖尿病并发症
柚皮苷%人臍靜脈內皮細胞%單覈細胞%粘附分子%活性氧%轉錄因子%糖尿病併髮癥
유피감%인제정맥내피세포%단핵세포%점부분자%활성양%전록인자%당뇨병병발증
naringin%human umbilicalvein endothelialcell%monocytes%adhesion molecule%reactive oxygen species%transcriptional factor%diabetic complications
目的 研究柚皮苷对高糖诱导的脐静脉内皮细胞(HUVECs)与单核细胞的粘附的抑制作用及机制.方法 分离培养HUVECs后,采用不同剂量的柚皮苷预处理HUVECs,然后高糖诱导HUVECs;加入荧光染料标记单核细胞系THP-1细胞与HUVECs共同孵育,荧光显微镜下观察HUVECs与单核细胞的粘附作用并拍照,比较粘附密度;提取总蛋白后,Westem blotting检测处理后HUVECs粘附分子的表达;荧光染料结合法检测处理后HUVECs活性氧的产生;提取处理的脐静脉内皮细胞核蛋白后,Western blotting检测核因子-κB在核内的量.结果 高糖诱导HUVECs与单核细胞的粘附,柚皮苷可显著抑制高糖诱导的HUVECs与单核细胞的粘附.柚皮苷抑制高糖诱导的HUVECs细胞间粘附分子和血管内皮细胞粘附分子的表达,柚皮苷抑制高糖诱导的HUVECs活性氧产生,柚皮苷抑制高糖诱导的HUVECs核因子-κB p65活化.结论柚皮苷可能通过抑制高糖诱导的HUVECs活性氧产生,从而抑制核因子-κB的活化,进一步抑制细胞间粘附分子和血管内皮细胞粘附分子的表达,影响HUVECs与THP-1细胞的粘附,从而抑制高糖诱导的血管炎症.
目的 研究柚皮苷對高糖誘導的臍靜脈內皮細胞(HUVECs)與單覈細胞的粘附的抑製作用及機製.方法 分離培養HUVECs後,採用不同劑量的柚皮苷預處理HUVECs,然後高糖誘導HUVECs;加入熒光染料標記單覈細胞繫THP-1細胞與HUVECs共同孵育,熒光顯微鏡下觀察HUVECs與單覈細胞的粘附作用併拍照,比較粘附密度;提取總蛋白後,Westem blotting檢測處理後HUVECs粘附分子的錶達;熒光染料結閤法檢測處理後HUVECs活性氧的產生;提取處理的臍靜脈內皮細胞覈蛋白後,Western blotting檢測覈因子-κB在覈內的量.結果 高糖誘導HUVECs與單覈細胞的粘附,柚皮苷可顯著抑製高糖誘導的HUVECs與單覈細胞的粘附.柚皮苷抑製高糖誘導的HUVECs細胞間粘附分子和血管內皮細胞粘附分子的錶達,柚皮苷抑製高糖誘導的HUVECs活性氧產生,柚皮苷抑製高糖誘導的HUVECs覈因子-κB p65活化.結論柚皮苷可能通過抑製高糖誘導的HUVECs活性氧產生,從而抑製覈因子-κB的活化,進一步抑製細胞間粘附分子和血管內皮細胞粘附分子的錶達,影響HUVECs與THP-1細胞的粘附,從而抑製高糖誘導的血管炎癥.
목적 연구유피감대고당유도적제정맥내피세포(HUVECs)여단핵세포적점부적억제작용급궤제.방법 분리배양HUVECs후,채용불동제량적유피감예처리HUVECs,연후고당유도HUVECs;가입형광염료표기단핵세포계THP-1세포여HUVECs공동부육,형광현미경하관찰HUVECs여단핵세포적점부작용병박조,비교점부밀도;제취총단백후,Westem blotting검측처리후HUVECs점부분자적표체;형광염료결합법검측처리후HUVECs활성양적산생;제취처리적제정맥내피세포핵단백후,Western blotting검측핵인자-κB재핵내적량.결과 고당유도HUVECs여단핵세포적점부,유피감가현저억제고당유도적HUVECs여단핵세포적점부.유피감억제고당유도적HUVECs세포간점부분자화혈관내피세포점부분자적표체,유피감억제고당유도적HUVECs활성양산생,유피감억제고당유도적HUVECs핵인자-κB p65활화.결론유피감가능통과억제고당유도적HUVECs활성양산생,종이억제핵인자-κB적활화,진일보억제세포간점부분자화혈관내피세포점부분자적표체,영향HUVECs여THP-1세포적점부,종이억제고당유도적혈관염증.
Objective To investigate the inhibitory effect of naringin on monocyte adhesion to high glucose-induced human umbilical vein endothelial cells (HUVECs). Methods Cultured HUVECs isolated from human umbilical cords were pretreated with or without naringin and induced with high glucose (33 mmol/L) for 48 h. Human monocyte THP-1 cells, after labeling with BCECF-AM, were co-cultured with the HUVECs for 30 min. The labeled THP-1 cells adhering to HUVECs were observed under fluoroscence microscope, and the inhibitory effect of naringin on the cell adhesion was evaluated by measuring the adhering cell density. Western blot analysis was used to detect the expressions of the adhesion molecules in the HUVECs, and reactive oxygen species (ROS) production in the HUVECs was measured using an oxidation-sensitive fluorescent probe (DCFH-DA). The nuclear extracts of the HUVECs were prepared to examine the expression of nuclear factor-kappa B (NF-κB) in the cellnuclei by Western blotting. Results HUVECs in high-glucose culture showed increased adhesion to THP-1 cells and enhanced expressions of the cell adhesion molecules, which were significantly attenuated by pretreatment with naringin (10-50 μg/ml). High glucose induced DCF-sensitive intracellular ROS production in the HUVECs,and this effect was inhibited by naringin pretreatment of the cells. Naringin also suppressed high glucose-induced increment of NF-κB expression in the cell nuclei of HUVECs. Conclusion Naringin can suppress high glucose-induced vascular inflammation possibly by inhibiting ROS production and NF-κB activation in HUVECs.
