阿托伐他汀钙对脂多糖诱导急性肺损伤小鼠核因子E2相关因子2/血红素加氧酶-1通路的影响

Effect of atorvastatin calcium on Nrf2/HO-1 pathway in mice with lipopolysaccharide-induced acute lung injury

目的探讨阿托伐他汀钙(AVT)对脂多糖(LPS)诱导急性肺损伤(ALI)小鼠核因子E2相关因子2/血红素加氧酶-1(Nrf2/HO-1)通路的影响。方法2021年9―12月,采用随机数字表法把15只BALB/c雌性小鼠分为NS组(尾静脉注射生理盐水)、LPS组(尾静脉注射LPS诱导ALI模型)、LPS+AVT组(经AVT灌胃治疗的ALI模型),每组各5只。收集动脉血,测定血氧分压(PaO_(2))、氧合指数(PaO_(2)/FiO_(2));处死后取出双肺,HE染色观察肺组织,测定湿干重比(W/D),酶联免疫吸附(ELISA)法测肺泡灌洗液(BALF)中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量,蛋白质印迹法(Western blotting)测肺组织中Nrf2、HO-1的表达。结果NS组、LPS组及LPS+AVT组PaO_(2)[(89.66±8.54)、(50.88±6.26)、(67.84±5.76)mmHg]、PaO_(2)/FiO_(2)[(426.95±40.67)、(242.29±29.79)、(323.05±27.42)mmHg]、W/D(3.12±0.54、4.95±0.60、3.85±0.46)、IL-1β[(37.95±6.18)、(115.09±12.42)、(75.61±7.89)ng/L]、TNF-α[(63.66±20.71)、(330.99±41.71)、(220.87±28.89)ng/L]、Nrf2(0.43±0.03、0.08±0.02、0.22±0.03)及HO-1(0.59±0.07、0.09±0.03、0.22±0.03)。与NS组相比,LPS组肺泡结构混乱不清,肺泡壁增厚,腔内炎性细胞大量渗出堆积,PaO_(2)及PaO_(2)/FiO_(2)降低,W/D比值增加,BALF IL-1β、TNF-α含量增多,肺组织Nrf2、HO-1蛋白表达相对降低(均P<0.05)。与LPS组相比,LPS+AVT组肺泡结构清晰度高,肺泡壁厚度及腔内炎性渗出改善,PaO_(2)及PaO_(2)/FiO_(2)升高,W/D比值降低,BALF IL-1β、TNF-α含量减少,肺组织Nrf2、HO-1蛋白含量增多(均P<0.05)。结论AVT可减轻LPS诱导ALI小鼠的炎症反应,这可能与激活Nrf2/HO-1通路有关。

Objective To investigate the effect of atorvastatin calcium(AVT)on nuclear factor E2-related factor 2/heme oxygenase-1(Nrf2/HO-1)pathway in mice with lipopolysaccharide(LPS)-induced acute lung injury(ALI).Methods From September to December 2021,fifteen female BALB/c mice were assigned randomly into NS group(injection with normal saline),LPS group(ALI model inducted by intra-tail injection with LPS)and LPS+AVT group(ALI model treated by intragastric administration with AVT),with 5 mice in each group.The arterial blood was taken for detecting the partial pressure of oxygen(PaO_(2))and the oxygenation index(PaO_(2)/FiO_(2)).The lung tissues were observed by HE staining and detected for wet/dry ratio(W/D).Enzyme linked immunosorbent assay(ELISA)was used to measure the expression of Interleukin-1β(IL-1β),Tumor necrosis factor-α(TNF-α)in bronchoalveolar lavage fluid(BALF)and western blot was used to proteins of Nrf2 and HO-1 in lung tissues.Results PaO_(2)inNS group,LPS group and LPS+AVT group was respectively[(89.66±8.54),(50.88±6.26),(67.84±5.76)mmHg];PaO_(2)/FiO_(2)was[(426.95±40.67),(242.29±29.79),(323.05±27.42)mmHg];W/D was(3.12±0.54,4.95±0.60,3.85±0.46);IL-1βwas[(37.95±6.18),(115.09±12.42),(75.61±7.89)ng/L];TNF-αwas[(63.66±20.71),(330.99±41.71),(220.87±28.89)ng/L];Nrf2 was(0.43±0.03,0.08±0.02,0.22±0.03)and HO-1 was(0.59±0.07,0.09±0.03,0.22±0.03).Compared with NS group,alveolar structure in LPS group was distorted,alveolar septa was thickened,alveolar spaces were accumulated with inflammatory cells;the levels of PaO_(2),PaO_(2)/FiO_(2),and proteins of Nrf2 and HO-1 in pulmonary tissue were decresed(P<0.05),while W/D ratio,IL-1βand TNF-αin BALF were increased(P<0.05).Compared with the LPS group,structure distortion,alveolar septa thickening and inflammatory cells accumulation in LPS+AVT group were reduced;the levels of PaO_(2),PaO_(2)/FiO_(2),and proteins of Nrf2 and HO-1 in lung tissues were increased(P<0.05),while W/D ratio,IL-1βand TNF-αin BALF were decreased(P<0.05).Conclusion AVT can reduce the inflammatory response in LPS-induced ALI mice,which may be related to the activation of Nrf2/HO-1 pathway.