细胞培养上清的收集.docVIP

消可宁对葡萄糖诱导血管内皮细胞诱生型一氧化氮合酶表达变化的抑制作用 ???? 【摘要】 目的 探讨消可宁对葡萄糖（GS）诱导血管内皮细胞诱生型一氧化氮合酶（iNOS）表达变化的影响。方法? 培养人脐静脉内皮细胞株ECV304细胞，用 mol/L GS处理h，用生理盐水、10－７ mol/L CCK8和0１ mg/L GS+10－６、10－７、１０－８? mol/L CCK8处理16 h；用比色法检测培养液中一氧化氮(NO)含量、细胞NOS活性，免疫细胞化学及蛋白质免疫印迹法检测iNOS蛋白表达。结果? 与生理盐水处理的对照组比较，GS诱导培养液NO含量增多、细胞NOS活性增高、iNOS蛋白表达上调；CCK8剂量依赖性抑制GS的上述效应，而单独作用对iNOS蛋白表达、NOS活性和NO含量均无明显影响。结论? CCK8可以明显抑制GS引起ECV304细胞iNOS蛋白表达上调，减少NO生成。????? 【关键词】 消可宁；葡萄糖；一氧化氮合酶；血管内皮细胞 Inhibitory effect of cholecystokinin octapeptide on lipopolysaccharideelicited inducible nitric oxide synthase expression in vascular endothelial cells?Department of Forensic Medicine and Pathophysiology, Hebei Medical University, Shijiazhuang 050017, Hebei, China (YAN Jun works at Department of Pathophysiology, Nankai Hospital, Tianjin 300100, China)Corresponding author: GU Zhenyong (Email: zhenyong88@)????? 【Abstract】 Objective? To investigate the effect of cholecystokinin octapeptide on lipopolysaccharide (GS)elicited inducible nitric oxide synthase (iNOS) expression in vascular endothelial cells. Methods Human umbilical vein endothelial cell line (ECV304 cells) was stimulated with vehicle (normal saline) or GS in the presence (001, 01, 1 mg/L) or absence (01 mg/L) of CCK8 (10－6－10－8 mol/L). Nitric oxide (NO) level and cellular nitric oxide synthase (NOS) activity were determined with spectrophotometrically. The iNOS expression was detected with immunocytochemical technique and Western blot. Results? Compared with normal saline, GS significantly induced the upregulation of iNOS protein expression in the cultured ECV304 cells, and NOS activity in ECV304 cells and NO level in cultured media were increased. CCK8 obviously inhibited abovementioned effect of GS in a dosedependent manner. Whereas CCK8 alone did not showed effect on iNOS protein expression, NO level and cellular NOS activity as compared with those values when vehicle was used. Conclusion? CCK8 inhibite GSelicited iNOS expression and NO production in ECV304 cells.?????