摘要
目的 :探讨八肽胆囊收缩素 ( CCK 8)对脂多糖 ( LPS)诱导培养的肺动脉内皮细胞 ( BPAEC)凋亡的影响。方法 :培养的 BPAEC用 L PS、CCK 8、CCK 8非特异性受体阻断剂丙谷胺分别或共同处理后 ,继续培养 2 4小时。用流式细胞术和核荧光染色方法检测细胞凋亡 ,用生物化学方法检测培养上清中乳酸脱氢酶( L DH)活性和丙二醛 ( MDA)含量 ,用流式细胞术定量检测过氧亚硝基阴离子 ( ONOO-)含量。结果 :LPS可诱导 BPAEC凋亡和坏死明显增多 ,培养上清中 MDA含量增高 ;CCK 8抑制 BPAEC凋亡和 MDA含量的增高 ,此作用为 CCK 8受体非特异性阻断剂丙谷胺翻转 ;CCK 8可抑制 L PS诱导 BPAEC生成 ONOO-增多 ;CCK 8和丙谷胺对 LPS诱导的 BPAEC坏死无明显影响。结论 :CCK 8可抑制 L PS诱导的 BPAEC凋亡 ,此作用由其受体介导 ,并与 CCK
Objective:To investigate the effect of cholecystokinin octapeptide (CCK8) on lipopolysaccharide (LPS)induced apoptosis of cultured pulmonary artery endothelial cells (BPAEC).Methods:BPAEC were challenged with LPS,CCK8,proglumide and vehicle,respectively.Then the cells continued to be cultured for 24 hours.The cellular apoptosis was analyzed by flow cytometer and fluorecent stain,and the content of malondialdehyde (MDA) and lactate dehydrogenase(LDH) activity were measured by biochemical methods.The cellular necrosis was expressed by LDH activity in supernatant.The endothelialderived peroxynitrite (ONOO -),a strong oxidant resulted from the reaction of nitric oxide and superoxide,was measured by the assay of ONOO - marker molecule nitrotyrosine (NT) with flow cytometer.Results:LPS could significantly induce increased apoptosis and necrosis in BPAEC,and elevate MDA contents in supernatant.CCK8 reversed LPSinduced increase in apoptosis and MDA contents,however these effects were abolished by proglumide,a nonspecific antagonist of CCK8 receptors.In contrast,CCK8 and proglumide had minimal effect on LPSinduced LDH activity,indicating that there was no significant change of necrosis in BPAEC.CCK8 inhibited LPSinduced ONOO - generation in BPAEC.Conclusions:LPSinduced apoptosis of BPAEC could be inhibited by CCK8,and it might be mediated by its receptors.Antioxidation activity of CCK8 might also play a role.
出处
《中国危重病急救医学》
CAS
CSCD
2001年第12期724-737,共14页
Chinese Critical Care Medicine
基金
国家自然科学基金资助项目(N o.3 9870 3 17
N o.3 9570 3 0 4 )
关键词
八肽胆囊收缩素
内皮细胞
肺动脉
细胞凋亡
cholecystokinin octapeptide
endothelial cell
pulmonary artery
apoptosis
