PDF(Pubmed)
Abstract：
Cardiac dysfunction is an important component of multiple organ failure caused by sepsis, and an important cause of high mortality in patients with sepsis. Herein, we attempted to determine whether myo-inositol oxygenase (MIOX) has proinflammation enzyme in infection-induced cardiac dysfunction (IICD) and its underlying mechanism.
Patients with IICD were collected by our hospital. A mouse model of IICD was induced into male db/db mice by cecal ligation and puncture (CLP). All mice were injected with 20 μL of LV-MIOX or LV-control short hairpin RNA using a 0.5-mL insulin syringe. On the second day, all mice were induced by CLP. H9C2 cell was also induced with lipopolysaccharide and adenosine triphosphate. Quantitative analysis of messenger RNAs (mRNAs) and gene microarray hybridization was used to analyze the mRNA expression levels. Enzyme-linked immunosorbent assay, immunofluorescence, and Western blot analysis were used to analyze the protein expression levels.
The serum expressions of MIOX mRNA level in patients with IICD were upregulated compared to normal healthy volunteers. MIOX promoted inflammation levels in the in vitro model of IICD. Si-MIOX inhibited inflammation levels in the in vitro model of IICD. MIOX accelerated inflammation and cardiac dysfunction in infection-induced mice. MIOX interacted with NLR family pyrin domain containing 3 (NLRP3) protein to reduce the degradation of NLRP3. The inhibition of MIOX reversed the effects of NLRP3 in the in vitro model of cardiac dysfunction.
Taken together, these findings demonstrate that MIOX accelerates inflammation in the model of IICD, which may be, at least in part, attributable to NLRP3 activity by the suppression of NLRP3 degradation in IICD.
Patients with IICD were collected by our hospital. A mouse model of IICD was induced into male db/db mice by cecal ligation and puncture (CLP). All mice were injected with 20 μL of LV-MIOX or LV-control short hairpin RNA using a 0.5-mL insulin syringe. On the second day, all mice were induced by CLP. H9C2 cell was also induced with lipopolysaccharide and adenosine triphosphate. Quantitative analysis of messenger RNAs (mRNAs) and gene microarray hybridization was used to analyze the mRNA expression levels. Enzyme-linked immunosorbent assay, immunofluorescence, and Western blot analysis were used to analyze the protein expression levels.
The serum expressions of MIOX mRNA level in patients with IICD were upregulated compared to normal healthy volunteers. MIOX promoted inflammation levels in the in vitro model of IICD. Si-MIOX inhibited inflammation levels in the in vitro model of IICD. MIOX accelerated inflammation and cardiac dysfunction in infection-induced mice. MIOX interacted with NLR family pyrin domain containing 3 (NLRP3) protein to reduce the degradation of NLRP3. The inhibition of MIOX reversed the effects of NLRP3 in the in vitro model of cardiac dysfunction.
Taken together, these findings demonstrate that MIOX accelerates inflammation in the model of IICD, which may be, at least in part, attributable to NLRP3 activity by the suppression of NLRP3 degradation in IICD.
摘要：
背景：心功能不全是脓毒症引起的多器官功能衰竭的重要组成部分，和脓毒症患者高死亡率的重要原因。在这里，我们试图确定肌醇加氧酶(MIOX)在感染诱导的心功能不全(IICD)中是否具有促炎酶及其潜在机制.
方法：收集我院诊断为ICD的患者。通过盲肠结扎和穿孔(CLP)将IICD的小鼠模型诱导为雄性db/db小鼠。使用0.5-mL胰岛素注射器向所有小鼠注射20μLLV-MIOX或LV对照短发夹RNA。第二天,所有小鼠均采用CLP诱导。还用脂多糖和三磷酸腺苷诱导了H9C2细胞。使用信使RNA（mRNA）的定量分析和基因微阵列杂交来分析mRNA表达水平。酶联免疫吸附测定,免疫荧光，蛋白质印迹分析用于分析蛋白质表达水平。
结果：与正常健康志愿者相比，CICD患者血清中MIOXmRNA表达水平上调。在IICD的体外模型中,MIOX促进炎症水平。Si-MIOX抑制IICD体外模型中的炎症水平。MIOX加速感染诱导的小鼠的炎症和心脏功能障碍。MIOX与含NLR家族pyrin结构域的3(NLRP3)蛋白相互作用以减少NLRP3的降解。MIOX的抑制逆转了NLRP3在心脏功能障碍的体外模型中的作用。
结论：综合来看，这些发现表明MIOX加速了ICD模型中的炎症，可能是，至少在某种程度上,通过抑制NICD中的NLRP3降解可归因于NLRP3活性。
方法：收集我院诊断为ICD的患者。通过盲肠结扎和穿孔(CLP)将IICD的小鼠模型诱导为雄性db/db小鼠。使用0.5-mL胰岛素注射器向所有小鼠注射20μLLV-MIOX或LV对照短发夹RNA。第二天,所有小鼠均采用CLP诱导。还用脂多糖和三磷酸腺苷诱导了H9C2细胞。使用信使RNA（mRNA）的定量分析和基因微阵列杂交来分析mRNA表达水平。酶联免疫吸附测定,免疫荧光，蛋白质印迹分析用于分析蛋白质表达水平。
结果：与正常健康志愿者相比，CICD患者血清中MIOXmRNA表达水平上调。在IICD的体外模型中,MIOX促进炎症水平。Si-MIOX抑制IICD体外模型中的炎症水平。MIOX加速感染诱导的小鼠的炎症和心脏功能障碍。MIOX与含NLR家族pyrin结构域的3(NLRP3)蛋白相互作用以减少NLRP3的降解。MIOX的抑制逆转了NLRP3在心脏功能障碍的体外模型中的作用。
结论：综合来看，这些发现表明MIOX加速了ICD模型中的炎症，可能是，至少在某种程度上,通过抑制NICD中的NLRP3降解可归因于NLRP3活性。
