Yang Chunmei, Gao Chunlin, Yu Mingdong, Li Bo, Lu Yuechun, Lü Guoyi
Department of Anesthesiology, Second Hospital of Tianjin Medical University, Tianjin 300211, China.
Department of Anesthesiology, Second Hospital of Tianjin Medical University, Tianjin 300211, China. Email:
Zhonghua Yi Xue Za Zhi. 2014 Dec 2;94(44):3527-30.
To investigate the effect of dexmedetomidine pretreatment on hypoxia/reoxygenation-induced injury in human umbilical vein endothelial cells and the possible mechanism.
Cultured human umbilical vein endothelial cell-12 (HUVEC-12) were randomly divided into four groups(n = 24): group control (C), group dexmedetomidine (D), group hypoxia/reoxygenation (H/R), group hypoxia/reoxygenation plus dexmedetomidine (H/R+D). In groups D and H/R+D, Dexmedetomidine 50 µmol/L was added to the culture medium and the cells were incubated for 2 h. Then groups C and D were exposed to regular incubator and incubated for 12 h, groups H/R and H/R+D were incubated in an anaerobic chamber for 6 h and then returned to a regular incubator and incubated for 6 h. Cell growth conditions were observed under inverted microscope, the cell viability and apoptosis were measured by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry respectively. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of CHOP mRNA. Western blotting was used to detect the expression of CHOP and cleaved caspase-3 protein.
Compared with group C, the cell survival rate was significantly decreased, the expression of CHOP mRNA and CHOP protein, cleaved caspase-3 protein were up-regulated, the apoptotic rate was significantly increased in group H/R and group H/R+D (P < 0.05),No significant difference was found in group D (P > 0.05);compared with group H/R, the cell survival rate was significantly increased, the expression of CHOP mRNA and CHOP protein, cleaved caspase-3 protein were down-regulated, the apoptotic rate was decreased significantly in the group H/R+D (P < 0.05).
Dexmedetomidine pretreatment can effectively attenuate hypoxia/reoxygenation-induced apoptosis to human umbilical vein endothelial cell-12. The mechanism maybe related with down-regulating the expression of CHOP.
探讨右美托咪定预处理对人脐静脉内皮细胞缺氧/复氧损伤的影响及其可能机制。
将培养的人脐静脉内皮细胞-12（HUVEC-12）随机分为四组（n = 24）：对照组（C）、右美托咪定组（D）、缺氧/复氧组（H/R）、缺氧/复氧加右美托咪定组（H/R+D）。D组和H/R+D组在培养基中加入50 μmol/L右美托咪定，细胞孵育2 h。然后C组和D组置于常规培养箱中孵育12 h，H/R组和H/R+D组在厌氧箱中孵育6 h，再放回常规培养箱中孵育6 h。在倒置显微镜下观察细胞生长情况，分别用甲基噻唑基四氮唑（MTT）法和流式细胞术检测细胞活力和凋亡情况。采用逆转录聚合酶链反应（RT-PCR）检测CHOP mRNA的表达。采用蛋白质免疫印迹法检测CHOP和裂解的caspase-3蛋白的表达。
与C组比较，H/R组和H/R+D组细胞存活率明显降低，CHOP mRNA、CHOP蛋白、裂解的caspase-3蛋白表达上调，凋亡率明显升高（P < 0.05），D组差异无统计学意义（P > 0.05）；与H/R组比较，H/R+D组细胞存活率明显升高，CHOP mRNA、CHOP蛋白、裂解的caspase-3蛋白表达下调，凋亡率明显降低（P < 0.05）。
右美托咪定预处理可有效减轻缺氧/复氧诱导的人脐静脉内皮细胞-12凋亡。其机制可能与下调CHOP表达有关。
