纯化兔血清对氧磷酶1与传统方法对敌敌畏所致大鼠肝脏损伤的疗效比较

Use of Purified Rabbit Serum Paraoxonase 1 Compared with the Traditional Method of Curative Effect on Dichlorvos-induced Liver Injury in Rats

目的探讨纯化兔血清对氧磷酶1(PON1)对敌敌畏中毒大鼠肝损伤的疗效是否优于传统方法。方法将30只雄性SD大鼠随机分为5组:空白对照组(A组)、敌敌畏染毒组(B组)、传统治疗组(C组)、PON1治疗组(D组)、联合治疗组(E组),每组6只。B、C、D、E组均给予敌敌畏(9 mg/kg)腹腔注射染毒,C组在染毒后立即(<2 min)给予阿托品(10 mg/kg)+碘解磷定(45mg/kg)腹腔注射,D组在染毒前30 min给予PON1 9 600 U/kg尾静脉注射,E组染毒前30 min给予PON1 9 600 U/kg尾静脉注射,染毒后立即(<2 min)给予阿托品(10 mg/kg)+碘解磷定(45 mg/kg)腹腔注射,A组给予等量的生理盐水对照。各组于造模前(0min)、造模后10 min、30 min、1 h、2 h、4 h、6 h分别颈内动脉取血,运用ELISA方法检测血清乙酰胆碱酯酶活性。各组分别于造模后12 h取肝脏组织,HE染色观察不同组肝脏的病理改变,运用免疫组化及Western blotting检测不同组肝脏4-羟基壬烯醛(4-HNE)的定性及定量表达。结果 B组胆碱酯酶明显下降,光镜下出现肝细胞严重脂肪变、核固缩等病理改变,可见4-HNE高度表达;C、D、E组上述各指标改变均较B组轻,且各组与B组之间均存在统计学差异(P<0.05);D、E组病理改变较C组轻,上述指标与C组之间均有统计学差异(P<0.05);E组较D组病理改变稍轻,上述指标无统计学差异(P>0.05)。结论 PON1对急性敌敌畏中毒大鼠肝损伤发挥保护作用,且效果优于传统治疗。

Objective To explore whether the use of purified rabbit serum paraoxonase 1 （PON1） for the treatment of dichlorvos-induced liver injury in rats is superior to traditional method. Methods Thirty male SD rats were randomly divided into the followint 5 groups, with 6 rats in each group ： control group （A group）, dichlorvos group （B group）, traditional treatment group （C group）, PON1 treatment group （D group）, combined treatment group （E group ）. Rats in groups B, C, D and E were adminstered dichlorvos by intraperitoneal injection 9 mg/kg. In group C, atropine 10 mg/kg and iodine solution 45 mg/kg were injected intrapelitoneally within 2 rain ＇after dichlorvos administration. In group D, PON1 was injected intravenously at a dose of 9 600 U/kg, 30 rain prior to poisoning. In group E, PON1 was injected intravenously at a dose of 9 600 U/kg, 30 min prior to poisoning, followed by in ）ravenous injection of atropine 10 mg/kg and iodine solution 45 mg/kg within 2 rain after poisoning. Rats in A group received normal saline. Blood was collected at different time points to examine the acetyl cholinesterase （ AChE ） -levels by ELISA method. Liver tissue were collected at 12 hours after model establishment to observe the pathological changes. The expression of 4 hydroxy 2-nonenal （4-HNE） in the liver tissue was detected by immunohistochemistry and Western blotting. Results In group B, AChE levels decreased significandy, liver cells showed severe fatty degeneration, karyopyknosis and other pathological changes, and 4-HNE expression increased. The pathological changes of group D and group E were less obvious than those of group C, and the 4-HNE expression in the group D and group E were significantly different from that in the group C （P 〈 0.05）. Conclusion PON 1 plays a protective role in dichlorvos-induced liver injury in rats, and this protection is better than that offered by traditional treatment.