The Impact And Mechanism Of CVVH On Endoplasnic Reticulum Stress And Apoptosis Of Huvec Cells Which Induced By Serum Of Patients With Mods

AIM:To investigate the impact and mechanism of continuous veno-venous 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:10 patients with MODS who received CVVH and 10 healthy volunteers were involved in this study. This experiment was divided into four groups: healthy control,group of pre-CVVH,group of 6-hour-CVVH and group of 20-hour-CVVH. Serum of healthy volunteers and serum of patients with MODS at pre- CVVH,6-hour-CVVH and 20-hour-CVVH were used to induced HUVEC cells respectively in vitro for 24h. The cell apoptosis was examined by TUNEL assay,the intracellular GRP78 protein was observed by immunocytochemistry,the mRNA expression of GRP78, CHOP,TNF-α,IL-6 of cells were measured by RT-PCR, The level of Serum SOD,MDA was detected by colorimetric method, the level of TNF-α,IL-6 of serum was detected by ELISA. RESULTS:Compared with healty control, the group of pre-CVVH showed higher cellular apoptosis rate(p<0.01),the mRNA expression of GRP78,CHOP,TNF-α,IL-6 of cells increased significantly(p<0.01), protein expression of GRP78 increased obviously.Compared with group of pre-CVVH, cellular apoptosis rate decreased significantly(p<0.01)and the mRNA expression of GRP78,CHOP,TNF-α,IL-6 of cells decreased significantly(p<0.01)either in the group of 6-hour-CVVH or group of 20-hour-CVVH. The mRNA expression of GRP78,CHOP,TNF-α,IL-6 of cells in group of 20-hour-CVVH were significantly lower than those in group of 6-hour-CVVH(p<0.01). Immunocytochemistry showed protein expression of GRP78 in group of 6-hour-CVVH was significantly lower than that in group of pre-CVVH, protein expression of GRP78 in group of 20-hour-CVVH was significantly lower than that in group of 6-hour-CVVH. Compared with healthy control, the level of SOD of serum decreased significantly and the level of MDA of Serum increased significantly in the group of pre-CVVH(p<0.01), the level of TNF-α,IL-6 of serum also increased significantly in the group of pre-CVVH(p<0.01),after CVVH, the level of SOD of serum increased significantly and the level of MDA of serum decreased significantly(p<0.01), the level of TNF-α,IL-6 of serum also decreased significantly (p<0.01).CONCLUSION:Apoptosis of HUVEC cells induced by serum of patients with MODS was associated with ERS,CVVH can relieve redox state, remove inflammatory cytokine, relieve ERS, this may be one of mechanisms of CVVH relieve damage of HUVEC cells induced by serum of patients with MODS.