连续性血液滤过对多脏器功能衰竭患者血清诱导的血管内皮细胞凋亡及内质网应激的影响

周洪文; 甘华; 杜晓刚; 李正荣

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连续性血液滤过对多脏器功能衰竭患者血清诱导的血管内皮细胞凋亡及内质网应激的影响

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- 连续性血液滤过对多脏器功能衰竭患者血清诱导的血管内皮细胞凋亡及内质网应激的影响
- Impact of continuous veno-venous hemofiltration on endoplasmic reticulum stress and apoptosis of HUVEC cells induced by serum of patients with multiple organ dysfunction syndrome
- 陆军军医大学学报 2010年32卷第8期页码：805-809
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  中图分类号： R459.5;R590.5;R363
- 纸质出版：2010
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周洪文,甘华,杜晓刚,李正荣.连续性血液滤过对多脏器功能衰竭患者血清诱导的血管内皮细胞凋亡及内质网应激的影响[J].陆军军医大学学报(原第三军医大学学报),2010,32(08):805-809.

　Zhou Hongwen,Gan Hua,Du Xiaogang,Li Zhengrong.Impact of continuous veno-venous hemofiltration on endoplasmic reticulum stress and apoptosis of HUVEC cells induced by serum of patients with multiple organ dysfunction syndrome[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(08):805-809.
周洪文,甘华,杜晓刚,李正荣.连续性血液滤过对多脏器功能衰竭患者血清诱导的血管内皮细胞凋亡及内质网应激的影响[J].陆军军医大学学报(原第三军医大学学报),2010,32(08):805-809. DOI：

　Zhou Hongwen,Gan Hua,Du Xiaogang,Li Zhengrong.Impact of continuous veno-venous hemofiltration on endoplasmic reticulum stress and apoptosis of HUVEC cells induced by serum of patients with multiple organ dysfunction syndrome[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(08):805-809. DOI：

摘要

目的　探讨连续性静脉-静脉血液滤过(continuous veno-venous hemofiltration，CVVH)对多脏器功能衰竭（multiple organ dysfunction syndrome，MODS）患者血清体外诱导的脐静脉内皮细胞凋亡及内质网应激的影响及其机制。　方法　分别用10例健康志愿者的血清及10 例MODS患者CVVH治疗前、治疗6 h、治疗20 h的血清体外诱导人脐静脉内皮细胞（human umbilical vein endothelial cells

HUVEC）24 h，TUNEL法检测HUVEC细胞凋亡率，免疫荧光观察细胞GRP78蛋白表达，RT-PCR检测细胞GRP78、CHOP、TNF-α、IL-6 mRNA表达，比色法检测健康志愿者及CVVH治疗不同时间组血清中SOD活性、MDA含量。ELISA法检测健康志愿者及CVVH治疗不同时间组血清中TNF-α、IL-6含量。　结果　与健康对照组相比，CVVH治疗前组细胞凋亡率明显升高（P0.01），GRP78、CHOP、IL-6、TNF-α mRNA表达量也显著增高（P0.01）。与CVVH治疗前比较，CVVH治疗6、20 h组细胞凋亡率明显下降（P0.01），GRP78、CHOP、IL-6、TNF-α mRNA表达量明显减少（P0.01），治疗20 h组较治疗6 h组进一步减少（P0.05）。免疫荧光显示健康对照组GRP78蛋白表达量很少，CVVH治疗前组GRP78蛋白表达量明显高于健康对照组，治疗6 h组较治疗前组明显减少，治疗20 h组较治疗6 h 组进一步减少。与健康对照组比较，CVVH治疗前组血清SOD活性明显下降（P0.01），MDA含量明显增高（P0.01），血清中TNF-α、IL-6含量也明显增高（P0.01）。经过CVVH治疗，SOD活性明显升高（P0.01）， MDA含量明显下降（P0.01），TNF-α、IL-6含量也明显下降（P0.01）。　结论　MODS患者血清体外诱导的HUVEC细胞凋亡与内质网应激有关，CVVH治疗能减轻氧化应激，清除炎症介质，减轻内质网应激程度，从而减少内质网相关的凋亡。

Abstract

Objective　To investigate the effect and implying mechanism of continuous veno-hemofiltration (CVVH) on endoplasmic reticulum stress (ERS) and apoptosis of HUVEC cells which induced by serum of patients with multiple organ dysfunction syndrome (MODS). 　Methods　Ten patients with MODS who received CVVH and 10 healthy volunteers were involved in this study. This experiment was divided into 4 groups: healthy control

group of pre-CVVH

group of 6-hour-CVVH and group of 20-hour-CVVH. Serum of healthy volunteers and the sera of patients with MODS at pre-CVVH

6-hour-CVVH and 20-hour-CVVH were used to induced HUVEC cells respectively in vitro for 24 h. The cell apoptosis was examined by TUNEL assay

the intracellular GRP78 protein was observed by immunocytochemistry

and the mRNA expressions of GRP78

CHOP

TNF-α

and IL-6 in the cells were measured by RT-PCR. The levels of serum SOD and MDA were detected by colorimetric method

and those of of TNF-α and IL-6 of by ELISA. 　Results　Compared with healthy control

the group of pre-CVVH showed higher cellular apoptosis rate (P0.01)

the mRNA expressions of GRP78

CHOP

TNF-α

and IL-6 in this group of cells were increased significantly (P0.01)

so was the protein expression of GRP78 (P0.01). Compared with group of pre-CVVH

cellular apoptosis rate was decreased significantly (P0.01) and the mRNA expressions of GRP78

CHOP

TNF-α

and IL-6 were decreased significantly (P0.01)either in the group of 6-hour-CVVH or group of 20-hour-CVVH. The mRNA expression of GRP78

CHOP

TNF-α and IL-6 of cells in group of 20-hour-CVVH were significantly lower than those in group of 6-hour-CVVH (P0.05). Immunocytochemistry showed protein expression of GRP78 in group of 6-hour-CVVH was significantly lower than that in group of pre-CVVH

and in group of 20-hour-CVVH significantly lower than that in group of 6-hour-CVVH. Compared with healthy control

the serum level of SOD was decreased significantly and the level of MDA was increased significantly in the group of pre-CVVH (P0.01)

and the level of TNF-α and IL-6 was also increased significantly in the group of pre-CVVH (P0.01). After CVVH

the serum level of SOD was increased significantly and that of MDA was decreased significantly (P0.01)

and those of TNF-α and IL-6 were also decreased significantly (P0.01). 　Conclusion　Apoptosis of HUVEC cells induced by serum of patients with MODS is associated with ERS. CVVH can relieve redox state

remove inflammatory cytokine

and relieve ERS

and this may be one of mechanisms of CVVH relieving damage of HUVEC cells induced by serum from MODS patients.

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