title: [Effect of PON1 overexpression on mouse diaphragmatic muscle cells injury caused by acute dichlorvos poisoning].
abstract: OBJECTIVE
  To investigate the effect of paraoxonase1 (PON1) overexpression on mouse diaphragmatic muscle cells injury caused by acute dichlorvos poisoning.
  
  
  METHODS
  Mouse diaphragmatic muscle cells were cultured routinely and infected with overexpression lentivirus. Cells were divided into normal control group, DDVP group, LV-GFP + DDVP group, LV-PON1 + DDVP group. Cell viability was determined by CCK-8 assay. Flow cytometry was used to detect cell apoptosis. The mRNA and protein expression of PON1 and Nrf2 in mouse diaphragmatic muscle cells was measured by RT-PCR and Western blot. Enzyme-linked immunosorbent assay was used to determine levels of acetyl cholinesterase (AchE), heme oxygenase 1 (HO-1) and quinone oxidoreductase-1 (NQO-1) in mouse diaphragmatic muscle cells. The activity of superoxide dismutase (SOD) and catalase (CAT) as well as malondialdehyde (MDA) content in cells was measured by chemical colorimetry.
  
  
  RESULTS
  After induced by 0, 80, 160, 320, 640 µmol/L DDVP for 24 hours, the viability of mouse diaphragmatic muscle cells was (100 ± 3.82)%, (82.13 ± 2.60)%, (53.57 ± 5.05)%, (30.77 ± 3.30)%, (14.20 ± 2.19)% respectively, changing in a concentration-dependent manner (P < 0.05). After induced by 160 µmol/L DDVP for 0, 6, 12, 24 hours, the viability of mouse diaphragmatic muscle cells was (100.17 ± 2.74)%, (76.13 ± 6.01)%, (66.53 ± 3.55)%, (53.57 ± 5.05)%, changing in a time-dependent manner (P < 0.05). The PON1 protein level in LV-PON1 group was higher than that of blank control group (0.370 ± 0.015 vs 0.232 ± 0.004, 0.197 ± 0.015 vs 0.037 ± 0.003, P < 0.05). The cell viability of LV-PON1 group is higher than that of DDVP group at different time point after induction of DDVP (P < 0.05). After induced by DDVP for 24 hours, the cell apoptosis rate and MDA content in LV-PON1 group were lower than those of DDVP group (P < 0.05). While levels of AchE, PON1 and Nrf2 protein expression, SOD and CAT, HO-1 and NQO-1 were higher than those of DDVP group (P < 0.05).
  
  
  CONCLUSIONS
  The overexpression of PON1 could effectively alleviate AchE inhibition by DDVP and induce Nrf2 expression to exert antioxidant effect, thus protected the mouse diaphragmatic muscle cells.
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Wu
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Jue
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Yuehui
authors.initials: Y
authors.lastname: Hou
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Guangliang
authors.initials: G
authors.lastname: Hong
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Guangju
authors.initials: G
authors.lastname: Zhao
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Ge
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Qiaomeng
authors.initials: Q
authors.lastname: Qiu
authors.affiliation: Wenzhou Medical University, Wenzhou 325000, China; Email: lzq640815@163.com.
authors.forename: Zhongqiu
authors.initials: Z
authors.lastname: Lu
mesh_terms: D000818:Animals; D000975:Antioxidants; D017209:Apoptosis; D043303:Aryldialkylphosphatase; D002374:Catalase; D002478:Cells, Cultured; D003964:Diaphragm; D004006:Dichlorvos; D051547:Heme Oxygenase-1; D008315:Malondialdehyde; D008565:Membrane Proteins; D051379:Mice; D032342:Muscle Cells; D013482:Superoxide Dismutase
chemical_list: D000975:Antioxidants; D008565:Membrane Proteins; D008315:Malondialdehyde; D004006:Dichlorvos; D002374:Catalase; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; D013482:Superoxide Dismutase; D043303:Aryldialkylphosphatase; C560867:PON1 protein, mouse