丙泊酚减弱CCL4诱导脑微血管内皮细胞促炎和细胞毒作用机制研究  

Mechanism of propofol attenuating CCL4-induced inflammation and cytotoxicity in cerebralmicrovascular endothelial cells

作　　者：赵娟 龚葛松 ZHAO Juan;GONG Gesong(Department of Anesthesiology,Qidong Hospital,Nantong University,Nantong,Jiangsu 226200,China;Department of Neurology,Qidong Hospital,Nantong University,Nantong,Jiangsu 226200,China)

机构地区：[1]南通大学附属启东医院麻醉科,江苏南通226200 [2]南通大学附属启东医院神经内科,江苏南通226200

出　　处：《国际检验医学杂志》2024年第8期996-1000,共5页International Journal of Laboratory Medicine

摘　　要：目的 探讨趋化因子配体4(CCL4)诱导脑微血管内皮细胞炎症和细胞毒作用,以及丙泊酚对CCL4诱导效应的拮抗作用。方法 将体外永生化人脑微血管内皮细胞hCMEC/D3分为为对照组、CCL4组、丙泊酚+CCL4组细胞。采用CCK-8试验检测各组细胞增殖活力,细胞克隆形成实验分析细胞克隆形成能力,流式细胞术Annexin V/PI双染检测细胞凋亡情况。采用实时荧光定量PCR(qPCR)检测细胞细胞炎症基因、血栓形成相关基因的表达情况,酶活性检测试剂盒检测环氧化酶-2(COX-2)及凝血酶活性。结果 与对照组细胞比较,CCL4组细胞增殖活力显著减弱,细胞克隆形成能力减低,凋亡细胞增加,差异有统计学意义(P<0.001);与CCL4组,丙泊酚+CCL4组细胞增殖活力显著上调,细胞克隆形成能力增加,凋亡细胞减少,差异有统计学意义(P<0.001)。qPCR检测结果显示,与对照组比较,CCL4组炎症相关基因白细胞介素(IL)-1β、IL-6、IL-8、核转录因子κB(NF-κB)、肿瘤坏死因子α(TNF-α)及COX-2基因表达上调,血栓形成基因Thrombin、Factor 8、VWF及TXA表达增加,COX-2酶及凝血酶活性增加,差异有统计学意义(P<0.001)。丙泊酚可明显抑制CCL4对hCMEC/D3的炎症相关基因及血栓形成基因的促进表达作用,以及CCL4对hCMEC/D3的COX2酶及凝血酶活性。结论 丙泊酚可减轻CCL4对脑微血管内皮细胞的细胞毒作用,并拮抗其促炎和促血栓形成基因表达。Objective To investigate the inflammatory and cytotoxic effects of CCL4-induced brain microvascular endothelial cells,and to analyze the antagonistic effect of propofol on CCL4-induced effects.Methods In vitro immortalized human brain microvascular endothelial cells hCMEC/D3 were divided into control group,CCL4 group and propofol+CCL4 group.Cell proliferation activity was detected by CCK-8,cell cloning ability was analyzed by cell cloning assay,and apoptosis was detected by flow Annexin V/PI double staining.Reverse transcription real-time fluorescence quantitative PCR(qPCR)was used to detect the expression of inflammatory genes and thrombosis-related genes,and enzyme activity detection kit was used to detect COX2 and thrombin activities.Results Compared with the control group,cell proliferation activity in CCL4 group was significantly decreased,cell cloning ability was decreased,and apoptosis was increased,and the differences were statistically significant(P<0.001).Compared with CCL4 group,the cell proliferation activity of propofol+CCL4 group was significantly up-regulated,the cell cloning ability was increased,and the apoptotic cells were decreased,with statistical significance(P<0.001).The results of qPCR showed that compared with the control group,the expressions of inflammatory genes IL-1β,IL-6,IL-8,NF-κB,TNF-αand COX-2 genes were upregulated in CCL4 group,and Thrombin,Factor 8,VWF and TXA expressions were increased,the activities of COX2 enzyme and thrombin were increased,and the difference was statistically significant(P<0.001).Propofol can significantly inhibit the promoting expression of inflammation related genes and thrombosis genes of hCMEC/D3 by CCL4,and the COX2 enzyme and thrombin activity of CCL4 on hCMEC/D3.Conclusion Propofol could reduce the cytotoxic effect of CCL4 on cerebral microvascular endothelial cells and antagonize the expression of proinflammatory and prothrombous genes.

关 键 词：CCL4 丙泊酚 人脑微血管内皮细胞 细胞增殖 凋亡 炎症 血栓 

分 类 号：R932[医药卫生—生药学] R446.6[医药卫生—药学]