2-十一烷酮通过调控重组与合成蛋白/硫氧还蛋白互作蛋白/硫氧化还原蛋白信号改善小鼠哮喘的研究

Research of 2-undecanone on improving asthmatic mice by regulating recombinant versus syntaxin/thioredoxin interacting protein/thioredoxin signal

目的研究2-十一烷酮对实验性哮喘小鼠重组与合成蛋白/硫氧还蛋白互作蛋白/硫氧化还原蛋白(Nrf2/TXNIP/TrX)信号的影响。方法将Balb/c小鼠随机分为正常组、模型组、对照组、实验组和联合组,每组6只。正常组未行造模,其余4组给予抗原混合液200μL致敏+1%OVA激发建模。在此基础上,对照组给予400 mg·kg^(-1)2-十一烷酮;实验组给予30 mg·kg^(-1)ML385;联合组给予400 mg·kg^(-1)2-十一烷酮+30 mg·kg^(-1)ML385;正常组和模型组均给予等量0.9%NaCl。用苏木精-伊红染色法观察肺组织炎性细胞的浸润情况,用酶联免疫吸附试验法检测血清免疫球蛋白E(IgE)和支气管肺泡灌洗液(BALF)中白细胞介素1β(IL-1β)水平,用蛋白质印迹法检测Nrf2、TXNIP、TrX和NOD样受体热蛋白结构域相关蛋白3(NLRP3)的表达水平。结果实验组、对照组、联合组、模型组和正常组的炎症评分分别为(4.33±0.65)、(1.00±0.41)、(3.00±1.21)、(3.33±0.32)和(0.67±0.31)分,血清IgE分别为(24.17±4.59)、(87.51±9.91)、(49.43±8.49)、(129.38±13.75)和(78.00±11.35)ng·mL^(-1),BALF中IL-1β分别为(44.17±7.11)、(154.45±17.75)、(87.04±15.72)、(216.29±19.64)和(134.87±23.83)pg·mL^(-1),Nrf2蛋白相对表达水平分别为1.00±0.00、0.46±0.02、0.71±0.03、0.30±0.02和0.48±0.02,TXNIP蛋白相对表达水平分别为1.00±0.00、4.16±0.57、1.55±0.21、5.25±0.18和3.59±0.24,TrX蛋白相对表达水平分别为1.00±0.00、0.38±0.02、0.79±0.01、0.24±0.03和0.42±0.02,NLRP3蛋白相对表达水平分别为1.00±0.00、4.33±0.42、2.26±0.31、5.21±0.44和3.67±0.29。实验组的上述指标与模型组比较,差异均有统计学意义(均P<0.05)。结论2-十一烷酮对OVA诱导的小鼠哮喘具有明显的治疗作用,其机制可能与调节Nrf2/TXNIP/TrX信号,抑制NLRP3炎症小体活化有关。

Objective To study the effect of 2-undecanone on the signal of recombinant and synthetic protein/thioredoxin interacting protein/thioredoxin(Nrf2/TXNIP/Tr X)in experimental asthma mice.Methods The Balb/c mice were randomly divided into normal,model,control,experimental and combined groups,with 6 mice in each group.The normal group did not make the model,and the other four groups were given antigen mixture 200μL sensitization+1%OVA excitation was used for modeling.On this basis,the control group was given 400 mg·kg^(-1)2-undecanone;the experimental group was given 30 mg·kg^(-1)ML385;the combined group was given 400 mg·kg^(-1)2-undecanone+30 mg·kg^(-1)ML385;the normal and model groups were given equal amounts of 0.9%of Na Cl.Hematoxylin eosin staining was used to observe the infiltration of inflammatory cells in lung tissue,and enzyme-linked immunosorbent assay was used to detect serum immunoglobulin E(Ig E)and interleukin-1β(IL-1β)in bronchoalveolar lavage fluid(BALF).The expression levels of Nrf2,TXNIP,Tr X and NOD-like receptor thermal protein domain associated protein 3(NLRP3)were detected by Western blot.Results The inflammatory scores of the experimental,control,combined,model and normal groups were(4.33±0.65),(1.00±0.41),(3.00±1.21),(3.33±0.32)and(0.67±0.31)points;serum Ig E levels were(24.17±4.59),(87.51±9.91),(49.43±8.49),(129.38±13.75)and(78.00±11.35)ng·mL^(-1);IL-1βlevels in BALFwere(44.17±7.11),(154.45±17.75),(87.04±15.72),(216.29±19.64)and(134.87±23.83)pg·mL^(-1);the relative expression levels of Nrf2 protein were 1.00±0.00,0.46±0.02,0.71±0.03,0.30±0.02and 0.48±0.02;the relative expression levels of TXNIP protein were 1.00±0.00,4.16±0.57,1.55±0.21,5.25±0.18 and 3.59±0.24;the relative expression levels of Tr X protein were 1.00±0.00,0.38±0.02,0.79±0.01,0.24±0.03 and 0.42±0.02;the relative expression levels of NLRP3 protein were 1.00±0.00,4.33±0.42,2.26±0.31,5.21±0.44 and 3.67±0.29,respectively.Comparing with the model group,the above indexes in the experimental group were statistically significant(all P<0.05).Conclusion 2-undecanone has obvious therapeutic effect on OVA induced asthma in mice,and its mechanism may be related to regulating Nrf2/TXNIP/Tr X signal and inhibiting the activation of NLRP3 inflammatory bodies.