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目的探讨高容量血液滤过(HVHF)治疗多脏器功能障碍综合征(multiple organ dysfunction syndrome,MODS)对内质网应激相关性细胞凋亡及应激标志蛋白Caspase-12基因水平及表达的影响,并探讨高容量血液滤过(high volume hemofiltration,HVHF)治疗MODS的机制。方法经二次打击建立犬MODS模型。分HVHF组和MODS组,于术前(T1)、内毒素注射完后0h(T2)、6h(T3)、12h(T4)及24h(T5)留血标本。体内实验:应用荧光定量PCR法检测肝、肾及肺组织Caspase-12m RNA基因表达水平。体外实验:2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Caspase-12m RNA基因表达、蛋白表达及内皮细胞凋亡率。结果体内实验:与MODS组相比,HVHF组肾、肺组织Caspase-12m RNA基因表达水平明显降低,差异有统计学意义(P<0.05),而肝组织Caspase-12m RNA基因表达水平无明显差异(P>0.05)。体外实验:干扰前与MODS组相比,HVHF组在T2-T5HUVECs Caspase-12m RNA基因表达水平、Caspase-12蛋白表达水平及细胞凋亡率均明显降低,差异有统计学意义(P<0.01)。干扰后2组HUVECs Caspase-12m RNA基因表达水平、Caspase-12蛋白表达水平及细胞凋亡率均较干扰前显著降低,差异有统计学意义(P<0.01)。结论内质网应激凋亡信号通路在MODS演进过程中占重要地位,经过HVHF治疗后内质网应激(ERS)凋亡信号通路中的关键蛋白Caspase-12基因表达、Caspase-12蛋白表达水平及细胞凋亡率均明显下降,此作用有助于减轻机体细胞凋亡程度,有效地救治MODS。而RNA干扰(RNAi)技术有望成为治疗MODS新的治疗靶点。
Objective To investigate the effect of high-volume hemofiltration (HVHF) on multiple organ dysfunction syndrome (MODS) on the expression of stress-related apoptosis and the expression of Caspase-12 in endoplasmic reticulum And to explore the mechanism of high volume hemofiltration (HVHF) treatment of MODS. Methods The canine MODS model was established by the second strike. Divide the HVHF group and the MODS group, preoperative (T1), endotoxin injection after 0h (T2), 6h (T3), 12h (T4) and 24h (T5) In vivo experiments: Quantitative real-time PCR was used to detect the gene expression of Caspase-12 mRNA in liver, kidney and lung. In vitro experiments: Two groups of human serum induced human umbilical vein endothelial cells (HUVECs) in vitro to establish ER stress-induced apoptosis model. Caspase-12 mRNA expression, protein expression and endothelial cell apoptosis rate were determined before and after si RNA interference. Results In vivo, the expression of Caspase-12 mRNA in kidney and lung tissues of HVHF group was significantly lower than that of MODS group (P <0.05), while the expression of Caspase-12 mRNA in liver tissue was not significantly different (P> 0.05). In vitro, Caspase-12 mRNA expression, Caspase-12 protein expression and apoptosis rate in HVHF group were significantly lower than those in MODS group (P <0.01) . Caspase-12 mRNA expression, Caspase-12 protein expression and apoptosis rate of HUVECs in two groups after interference were significantly lower than those before interference (P <0.01). Conclusion Endoplasmic reticulum stress apoptosis signaling pathway plays an important role in the development of MODS. After HVHF treatment, the expressions of Caspase-12, Caspase-12 and Caspase-12 in ERS apoptotic signal pathway Levels and apoptosis rates were significantly decreased, this effect helps to reduce the degree of apoptosis in the body, effective treatment of MODS. RNA interference (RNAi) technology is expected to become a new therapeutic target for the treatment of MODS.