血液净化治疗对多脏器功能障碍综合征犬内质网应激信号通路作用机制的研究

发布时间：2019-02-26 19:01

【摘要】：目的探讨高容量血液滤过(HVHF)对多脏器功能障碍综合征(MODS)犬内质网应激标志蛋白JNK2 m RNA表达的影响。方法采用二次打击建立犬MODS模型,12只犬随机分为MODS组(n=6)和HVHF组(n=6),HVHF组给予HVHF治疗24 h。于术前(T1),内毒素注射完后0 h(T2)、6 h(T3)、12 h(T4)及24 h(T5)留取血标本。体内实验:检测肝、肺、肾组织细胞JNK2 m RNA水平。体外实验:两组血清体外诱导人脐静脉内皮细胞(HUVECs)建立内质网应激凋亡模型。测定si RNA干扰前后JNK2 m RNA表达、蛋白表达及内皮细胞凋亡率。结果体内实验:HVHF组肝脏JNK2 m RNA水平高于MODS组(P0.01)、肺脏明显低于MODS组(P0.01)。体外实验:(1)JNK2 m RNA表达:与T1时间相比,HVHF组在T3~T5 JNK2 m RNA水平明显降低,差异有统计学意义(P0.01);与si RNA干扰前相比,干扰后两组JNK2 m RNA表达水平均明显下降,差异具有统计学意义(P0.01);(2)JNK蛋白表达:与T1时间相比,HVHF组在T3~T5JNK蛋白表达明显降低,差异有统计学意义(P0.01);与MODS组相比,HVHF组在T3~T5 JNK蛋白表达明显降低,差异有统计学意义(P0.01);与si RNA干扰前相比,干扰后两组JNK蛋白表达均明显下降,差异具有统计学意义(P0.01);(3)内皮细胞凋亡率:与T1时间相比,HVHF组在T3~T5内皮细胞凋亡率明显降低,差异有统计学意义(P0.01);与MODS组相比,HVHF组在T3~T5内皮细胞凋亡率明显降低,差异有统计学意义(P0.01);与si RNA干扰前相比,干扰后两组内皮细胞凋亡率均明显下降,差异具有统计学意义(P0.01)。结论 JNK信号通路介导了MODS犬过度内质网应激时的细胞凋亡,HVHF可通过清除炎症介质,减少氧化应激,减轻内质网应激,减少细胞凋亡。
[Abstract]:Aim to investigate the effect of high volume hemofiltration (HVHF) on the expression of endoplasmic reticulum stress marker JNK2 m RNA in dogs with multiple organ dysfunction syndrome (MODS). Methods 12 dogs were randomly divided into two groups: MODS group (n = 6) and HVHF group (n = 6), HVHF) treated with HVHF for 24 h. The MODS model of dogs was established by twice beating. Blood samples were collected before operation (T1), 0 h (T 2), 6 h (T 3), 12 h (T 4) and 24 h (T 5) after endotoxin injection. In vivo experiment: the levels of JNK2 m RNA in liver, lung and kidney were measured. In vitro experiment: two groups of serum induced human umbilical vein endothelial cell (HUVECs) in vitro to establish endoplasmic reticulum stress apoptosis model. The expression of JNK2 m RNA, the expression of protein and the apoptosis rate of endothelial cells were measured before and after si RNA interference. Results in vivo, the level of JNK2 m RNA in liver of HVHF group was higher than that of MODS group (P0.01), and the level of lung was significantly lower than that of MODS group (P0.01). In vitro experiment: (1) JNK2 m RNA expression: compared with T1 time, the level of T3~T5 JNK2 m RNA in HVHF group was significantly lower than that in T1 group (P0.01). Compared with that before si RNA interference, the expression level of JNK2 m RNA in the two groups was significantly decreased after the interference (P0.01). (2) the expression of JNK protein: compared with T1 time, the expression of T3~T5JNK protein in HVHF group was significantly lower than that in MODS group (P0.01), and the expression of T3~T5JNK protein in HVHF group was significantly lower than that in MODS group (P0.01). Compared with that before si RNA interference, the expression of JNK protein in the two groups decreased significantly after interference (P0.01). (3) apoptosis rate of endothelial cells: compared with T1 time, the apoptosis rate of T3~T5 in HVHF group was significantly lower than that in T1 group (P0.01). Compared with MODS group, the apoptosis rate of T3~T5 endothelial cells in HVHF group was significantly lower than that in T3~T5 group (P0.01). Compared with that before si RNA interference, the apoptosis rate of endothelial cells in the two groups decreased significantly after interference, the difference was statistically significant (P0.01). Conclusion JNK signaling pathway mediates apoptosis during excessive endoplasmic reticulum stress in MODS dogs. HVHF can eliminate inflammatory mediators, reduce oxidative stress, relieve endoplasmic reticulum stress and reduce apoptosis.
【作者单位】：新疆医科大学第一附属医院肾病科;霍城县第一人民医院内科;
【基金】：新疆维吾尔自治区自然科学基金(2012211A065)
【分类号】：R459.7

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