中国中西医结合外科杂志2017，Vol.23Issue(6)：633-637,5.DOI:10.3969/j.issn.1007-6948.2017.06.014
重型胰腺炎大鼠血浆线粒体DNA变化与器官损伤的关系
Relationship between Plasma Mitochondrial DNA and Organ Damage in Rat Model of Severe Acute Pancreatitis
摘要
Abstract
Objective To detect the concentration of plasma mitochondrial DNA (mtDNA) in rat model of severe acute pancreatitis (SAP) and investigate the relationship between mtDNA level and organ damage.Methods Thirty healthy adult male Wistar rats were randomly divided into Sham operation group and SAP model group with 15 rats in each group.The rat SAP model was induced by retrogradely injection of 5 ％sodium taurocholate through the biliopancreatic duct.After 24 h,the whole blood was taken and the plasma was separated to measure the levels of amylase and lipase.The dissociative DNA was extracted from plasma and amplified by fluorescent quantitative PCR amplification using mitochondrial cytochrome B and cytochrome oxidase subunit Ⅲ primers.The level of mtDNA in circulating blood was represented by the PCR amplification product.The lung,liver and kidney tissues were collected.One part was used for HE staining,histopathological examination and scoring,and the other part was homogenized to extract the supematant and used to measure the levels of TNF-α,IL-1 and IL-6 by ELISA method.The two groups were compared using the index single factor variance analysis.Correlation analysis between the mtDNA level and pathological score was performed by Spearman rank correlation analysis.Results Compared with Sham operation group,the levels of amylase [(10444.5 ± 1863.2)U/L] and lipase [(1751.3 ± 121.3)U/L] in SAP model rats were significantly increased (P ＜ 0.01).The pathological scores and levels of TNF-α [(88.4 ± 15.4) pg/mg in lung,(79.3 ± 10.3) pg/mg in liver,(69.5 ± 9.6) pg/mg in kidney],IL-1 [(0.68 ± 0.11) μg/g in lung,(0.72 ± 0.17) μg/g in liver,(0.85 ± 0.17) μg/g in kidney] and IL-6 [(252.1 ± 18.7) pg/mg in lung,(107.5 ± 12.3) pg/mg in liver,(118.5 ± 15.2) pg/mg in kidney] were significantly increased (P ＜ 0.01).The circulating mtDNA level was also increased significantly and had a significant positive correlation with organ injury score (P ＜ 0.05).The relative expression of cytochrome B (0.4891 ± 0.0921) and cytochrome oxidase subunit Ⅲ (0.6082 ± 0.1142)in mitochondria of model group was increased significantly compared with that in Sham operation group (0.2856 ± 0.0878) and (0.1985 ± 0.0854),respectively) (P ＜ 0.01).The expression levels were positively correlated with the organ damage score.Conclusion The increase of mtDNA concentration in circulating blood of SAP model rats is related to the degree of organ damage.关键词
重型急性胰腺炎/线粒体DNA/损伤相关分子模式/细胞色素B/细胞色素C氧化酶Key words
Severe acute pancreatitis/mitochondrial DNA/damage associated molecular pattern/cytochrome B/cytochrome C oxidase分类
生物科学引用本文复制引用
刘韦鋆,余剑波,蔡旺,李东华,张一,聂卫,刘大卫,刘洪斌..重型胰腺炎大鼠血浆线粒体DNA变化与器官损伤的关系[J].中国中西医结合外科杂志,2017,23(6):633-637,5.基金项目
国家自然科学基金资助项目(81373854) （81373854）
