法舒地尔对急性乙酰甲胺磷中毒致心肌损伤的保护作用及机制

Protective effects and mechanisms of Fasudil on acute myocardial injury caused by acephate pesticide poisoning

目的探讨法舒地尔对急性乙酰甲胺磷中毒心肌损伤的保护作用及其机制。方法SD雄性大鼠随机分为4组:生理盐水组(A组)、中毒组(B组)、常规治疗组(C组)和法舒地尔联合常规治疗组(D组)。按时间点依次取全血及左心室部分组织为样本。酶联免疫吸附测定法(enzyme-linked immuno sorbent assay,ELISA)检测血清中肌钙蛋白I(roponin I,cTnI)和肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)的含量。免疫印迹法(western blot,WB)测定TNF-α、Ras同源基因家族A(Ras homolog gene family member A,RhoA)蛋白含量。免疫共沉淀(Co-Immunoprecipitation,Co-IP)和WB检测RhoA-GTP蛋白含量。苏木精-伊红染色法(hema-toxylin-eosin staining,HE染色法)观察心肌细胞组织学变化。结果法舒地尔可以降低中毒大鼠死亡率,与生理盐水组(A组)比较,中毒组(B组)TNF-α、cTnI水平明显升高,差异有统计学意义(P<0.05);D组与C组比较,D12组cTnI水平低于C12组,D12组和D24组TNF-α水平低于C12组和C24组,差异均有统计学意义(P<0.05)。法舒地尔可以显著改善心肌纤维排列,抑制炎性细胞浸润等。结论急性乙酰甲胺磷农药可诱导TNF-α升高,激活RhoA/ROCK信号通路,从而导致心肌损伤,而法舒地尔能够阻断RhoA/ROCK信号通路,起到对心肌损伤的保护作用。

Objective To investigate the protective effect of Fasudil on acute myocardial injury to acephate pesticide poisoning and its possible mechanism.Methods The rats were randomly divided into 4 groups:the normal saline group(Group A),poisoning group(Group B),routine treatment group(Group C),Fasudil combined with routine treatment group(Group D).According to the time,take some blood and left ventricular tissue.The concentration of serum cTnl was detected with ELISA.TNF-α,RhoA protein content was detected with Western blot.The level of RhoA-GTP protein was detected with immunoprecipitation.Results Fasudil can decrease the mortality of poisoning rats.Compared with group A,the levels of TNF-αand cTnl in group B was higher than that in group A.Compared the group D with group C,The levels of cTnl and TNF-α in group DI2 were lower than those in group C12.The levels of TNF-α in group D24 was lower than those in group C24.Fasudil significantly improved myocardial fiber arrangement and inhibited inflammatory cell infiltration.Conclusion AOPP may induce an increase in TNF-α and then further activate the RhoA/ROCK signaling pathway,resulting in myocardial damage.Fasudil as ROCK inhibitors,capable of blocking the RhoA/ROCK signaling pathway plays a protective effect on myocardial injury.