【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(P<0.01)、肺脏明显低于MODS组(P<0.01)。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(P<0.01);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(P<0.01);(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(P<0.01);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(P<0.01);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(P<0.01);(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(P<0.01);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(P<0.01);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(P<0.01)。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。更多还原

【Abstract】 Objective To investigate the expression of endoplasmic reticulum stress marker proteins JNK2 gene in dogs with multiple organ dysfunction syndrome(MODS) treated with high-volume hemofiltration treatment(HVHF). Methods Two-hit method was used to establish the dog MODS model.Twelve Beagle dogs were randomly divided into MODS group(n=6) and HVHF group(n=6). HVHF was performed about 24 hours after LPS injection in MODS dogs of HVHF group. Specimens from blood samples before operation(T1), before LPS injection and 0 h(T2), 6 h(T3), 12 h(T4), 24 h(T5) after LPS injected. In vivo tests: detecting JNK2 m RNA level in liver, lung and kidney tissue. In vitro tests: serum of two groups to induce human umbilical vein endothelial cells(HUVECs). In vitro to establish endoplasmic reticulum stress apoptosis models. Determined JNK2 m RNA expression, protein expression and endothelial cell apoptosis rate before and after si RNA interference. Results In vivo tests: JNK2 m RNA expression of liver in HVHF group were higher than MODS group(P<0.01). JNK2 m RNA expression of lung in HVHF group were was significantly lower than MODS group(P<0.01). In vitro tests:(1) JNK2 gene expression:compared with the T1 time, HVHF group was significantly lower in T3-T5 JNK2 m RNA level, the difference was statistically significant(P<0.01). Compared with the si RNA interference gave before, after interference the two group of JNK2 m RNA expression levels were significantly decreased, the difference was statistically significant(P<0.01);(2) JNK protein expression: compared with the T1 time, HVHF group was significantly lower in JNK protein expression at T3-T5, the difference was statistically significant(P<0.01); Compared with the MODS group, HVHF group was significantly lower in JNK protein expression at T3-T5, the difference was statistically significant(P<0.01); Compared with the si RNA interference gave before, two groups of JNK protein expression were significantly decreased after the disturbance, the difference was statistically significant(P<0.01);(3) Endothelial cell apoptosis rate:compared with the T1 time, HVHF group was significantly lower in the endothelial cell apoptosis rate at T3-T5, the difference was statistically significant(P<0.01); Compared with the MODS group, HVHF group was significantly lower in the endothelial cell apoptosis rate at T3-T5, the difference was statistically significant(P<0.01); Compared with the si RNA interference gave before, after interference the two groups were significantly decreased in the endothelial cell apoptosis rate, the difference was statistically significant(P<0.01). Conclusion JNK signaling pathway mediated apoptosis in MODS dogs over endoplasmic reticulum stress, HVHF can reduce apoptosis by clearing inflammatory mediators, reducing oxidative stress, and reducing the endoplasmic reticulum stress.更多还原