| 摘要: |
| 目的观察对氧磷酶1（PON1）过表达对敌敌畏中毒小鼠肝损伤的保护作用。方法将96只Balb/c小鼠按随机数字表法分为正常对照组、敌敌畏染毒组、对照慢病毒（Lv-GFP）＋敌敌畏组和PON1过表达重组慢病毒（Lv-PON1）＋敌敌畏组，每组24只。Lv-GFP＋敌敌畏组、Lv-PON1＋敌敌畏组小鼠分别经尾静脉转染Lv-GFP、Lv-PON1第3天，连同敌敌畏染毒组小鼠腹腔注射敌敌畏溶液9mg/kg染毒，同时正常对照组给予等量0.9%氯化钠注射液。染毒后分别于6、12、24、48h各取6只小鼠麻醉处死，化学比色法检测血清乙酰胆碱酯酶（AChE）、ALT和AST活力，ELISA测定肝组织TNF-α和IL-1β水平，Westernblot检测肝组织NF-κBp65表达，HE染色观察肝组织病理学变化。结果与正常对照组比较，6、12、24h敌敌畏染毒组AChE活力下降及6、12、24、48hALT、AST活力升高，差异均有统计学意义（均P＜0.05）；与相应时间点敌敌畏染毒组比较，Lv-PON1＋敌敌畏组AChE活力明显升高，ALT、AST活力明显降低（P＜0.05）。与正常对照组比较，6、12、24、48h敌敌畏染毒组TNF-α、IL-1β水平升高，NF-κBp65表达升高，差异均有统计学意义（均P＜0.05）；与相应时间点敌敌畏染毒组比较，Lv-PON1＋敌敌畏组TNF-α、IL-1β水平明显降低，NF-κBp65表达明显降低，除48hIL-1β外，差异均有统计学意义（均P＜0.05）。光镜下可见，敌敌畏染毒组染毒后12h肝病理损伤明显，Lv-PON1＋敌敌畏组小鼠肝损伤较之有所减轻。结论调控PON1过表达能够通过下调敌敌畏急性中毒小鼠肝组织NF-κBp65表达，减少TNF-α、IL-1β释放，减轻炎症损伤，对敌敌畏急性中毒小鼠肝损伤具有一定的保护作用。 |
| 关键词: 敌敌畏 对氧磷酶 1 肝损伤 炎症 |
| DOI： |
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| 基金项目:浙江省“十二五”重点学科建设计划资助；浙江省中医药重点学科建设计划；浙江省医学创新学科建设计划 |
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| Over-expression of PON1 protects mice from dichlorvos poisoning-induced liver injury |
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CHEN Xiaoyu, GUO Ruijuan, Wang Fei, WU Bin, HONG Guangliang, ZHAO Guangju, LI Mengfang, QIU Qiaomeng, LU Zhongqiu
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Huangyan Hospital of Wenzhou Medical University
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| Abstract: |
| Objective To investigate the effect of over expression of paraoxonase1 gene(PON1) on inflammation and liver injury induced by dichlorvos (DDVP) poisoning in mice. Methods Ninety six Balb/c mice were randomly divided into 4 groups: normal control group, the dichlorvos poisoning group (9 mg/kg dichlorvos), blank lentivirus group (Lv-GFP + dichlorvos) and PON1 recombinant lentivirus group (Lv-PON1+dichlorvos) with 24 mice in each group. The Lv-GFP, Lv-PON1 were transfected to mice via tail vein in Lv-GFP and Lv-PON1 groups and 9 mg/kg dichlorvos solution were intraperitoneal injected to mice 3 days after transfection, while same amount of saline solution was given to normal control group. At 6 h, 12 h, 24 h, 48 h after dichlorvos injection, 6 mice were sacrificed in batch. Serum acetylcholinesterase (AChE), alanine aminotransferase (ALT) and aspartate
transaminase (AST) were measured with chemical colorimetry. The levels of tumor necrosis-α (TNF-α) and interleukin 1β (IL -
1β) in liver tissue were determined by enzyme-linked immunosorbent (ELISA). The expression of NF - κBp65 in liver tissue was detected by Western blot. The pathological changes of liver tissue were observed with HE staining. Results Compared with normal control group, the AChE activity decreased and ALT, AST activity increased at 6 h, 12 h, 24 h after dichlorvos injection (P＜0.05). Compared to dichlorvos group at corresponding time points, the AChE activity of Lv-PON1+dichlorvos group was significantly increased, and ALT, AST activity significantly decreased(P＜0.05). Compared with normal control group, the TNF-α, IL-1β, and NF-κBp656 levels in dichlorvos group at 6 h, 12 h, 24 h, 48 h were elevated (P＜0.05). Compared with dichlorvos group at corresponding time points, the TNF-α, IL-1β levels and NF-κBp65 expression in Lv-PON1+dichlorvos group were significantly decreased (P＜0.05), expect the IL-1β expression in 48h. Compared with the Lv-PON1+dichlorvos group, liver injury in dichlorvos group at 12h were significantly severe. Conclusion Over expression of PON1 can reduce inflammation and protect the liver injury induced by acute dichlorvos poisoning in mice, which is associated with down-regulation of NF-κBp65 expression and reduced TNF-α, IL-1β levels. |
| Key words: Dichlorvos PON1 Liver injury Inflammation |
