新斯的明对百草枯中毒大鼠全身炎症反应和肺损伤的影响

Effect of neostigmine methylsulfate on paraquat induced inflammation and lung injury in rats

目的观察胆碱酯酶抑制剂——新斯的明对百草枯中毒大鼠全身炎症反应和肺损伤的影响。方法 96只SD大鼠随机分为3组(n=32)。NES组:腹腔注射百草枯(paraquat,PQ)溶液20 mg/kg后2 h腹腔注射新斯的明300μg/kg;PQ组:腹腔注射等量PQ溶液后2 h腹腔注射生理盐水300μg/kg;对照组:以等量生理盐水代替PQ溶液腹腔注射。在6、24、72 h 3个时间点观察大鼠血清及肺组织TNF-α、IL-6、IL-10变化;分批处死大鼠进行肺组织病理学观察以及测量肺湿/干质量(W/D);Real-Time PCR检测肺组织SOCS1、SOCS3 mRNA表达。结果 PQ染毒后,大鼠血清和肺组织TNF-α、IL-6、IL-10均显著升高,NES组在染毒后各时间点血清和肺组织TNF-α水平低于PQ组(P<0.05),在染毒后24、72 h血清及肺组织IL-6水平低于PQ组(P<0.05)。染毒后各时间点NES组IL-10水平高于PQ组(P<0.05)。染毒后72 h,病理学观察可见NES组大鼠肺损伤程度较PQ组轻。NES组24、72 h肺湿干质量比低于PQ组(P<0.05)。PQ染毒后各时间点SOCS1、SOCS3 mRNA表达水平高于对照组(P<0.05)。NES组SOCS3 mRNA表达水平高于PQ组,但SOCS1 mRNA表达水平与PQ组差异无统计学意义(P>0.05)。结论新斯的明能减轻PQ中毒大鼠的全身炎症反应和肺损伤程度。

Objective To investigate the effect of neostigmine methylsulfate on paraquat induced systemic inflammation and lung injury in a rat model. Methods Ninety-six rats were randomly divided into paraquat poisoning group （PQ group）, neostigmine group （NES group） and normal control group （NC group）. For the PQ and NES group, the rats were intraperitoneally injected with paraquat （20 mg/kg）, 2 h later followed by neostigmine methylsulfate （300 μg/kg） and saline, respectively. For the NC group, the rats were given equal volume of saline instead of paraquat. All rats were sacrificed at 6, 24 and 72 h after paraquat poisoning. Lung tissues were excised for pathological observation and wet to dry weight （W/D） ratio measurement. The levels of tumor necrosis factor （TNF）-α, interleukin （IL）-6, IL-10 in serum and lung tissues were detected at different time points. mRNA expression of suppressor of cytokine signaling 1 （SOCS1） and SOCS3 was detected by real-time polymerase chain reaction （RT-PCR）. Results Pathological observation showed that the lung injury in the NES group was milder than that in PQ group at 72 h. The lung W/D ratio at 24 and 72 hours were significantly lower in NES group compared to the PQ group （P〈0.05）. The levels of TNF-α, IL-6 and IL-10 in the serum and lung tissues remarkably increased after paraquat injection as compared to the control. The NES group had significantly lower levels of TNF-α （6, 24, 72 h） and IL-6 （24, 72 h） than the PQ group （P〈0.05）. In contrast, IL-10 level （6, 24, 72 h） were significantly higher in the NES versus the PQ group （P〈0.05）. In addition, the mRNA expression of SOCS1 and SOCS3 in lung was significantly increased in the PQ and NES groups compared with the NC group （P〈0.05）. SOCS3 expression was significantly higher in the NES group than the PQ group （P〈0.05）, but there was no significant difference in SOCS1 expression between the 2 groups. Conclusion Neostigmine methylsulfate has a protective effect on systemic inflammation and lung injury after paraquat poisoning, which is likely related to activation of cholinergic anti-inflammatory pathway.