非布索坦灌胃对百草枯诱导的大鼠肺损伤改善作用及其机制

Effect of intragastric administration of febuxostat on paraquat-induced lung injury in rats

目的观察非布索坦灌胃对百草枯诱导的大鼠肺损伤改善作用,并探讨其机制。方法21只大鼠随机分为3组,分别为对照组、百草枯组、百草枯+非布索坦组,百草枯组和百草枯+非布索坦组给予百草枯一次性灌胃染毒,百草枯+非布索坦组在染毒24 h后进行每天非布索坦灌胃治疗,对照组、百草枯组每天灌胃等体积生理盐水。各组大鼠灌胃14 d后处死,分离肺组织并心脏取血。采用HE染色法观察各组大鼠肺组织病理变化。采用比色法检测血清乳酸脱氢酶(LDH),采用ELISA法检测血清可溶性糖基化终产物受体(sRAGE)。采用比色法检测肺组织中氧化应激标志物黄嘌呤氧化酶(XO)、超氧化物歧化酶(SOD)、总抗氧化能力(TAC)、丙二醛(MDA)、羟脯氨酸。采用Western blotting法检测大鼠肺组织中PI3K、Akt、β-catenin蛋白,采用双抗体夹心ELISA法检测肺组织中基质金属蛋白酶-9(MMP-9)、白介素-8(IL-8)、血管内皮生长因子(VEGF)。结果对照组大鼠肺泡结构正常,百草枯组大鼠肺组织出现重度纤维组织增生、混合炎性细胞浸润和肺水肿情况,百草枯+非布索坦组大鼠肺组织中呈现中度纤维组织增生和炎性细胞浸润情况。与对照组相比,百草枯组血清LDH水平显著升高、sRAGE水平显著降低(P均<0.05);与百草枯组相比,百草枯+非布索坦组LDH水平显著降低、sRAGE水平显著升高(P均<0.05)。与对照组相比,百草枯组XO、MDA、羟脯氨酸水平均显著升高(P均<0.05),SOD、TAC水平均显著降低(P均<0.05);与百草枯组相比,百草枯+非布索坦组XO、MDA、羟脯氨酸水平均显著降低(P均<0.05),SOD、TAC水平均显著升高(P均<0.05)。与对照组相比,百草枯组大鼠肺组织中PI3K、Akt、β-catenin蛋白相对表达量和MMP-9、IL-8、VEGF水平均显著升高(P均<0.05);与百草枯组相比,百草枯+非布索坦组大鼠肺组织中PI3K、Akt、β-catenin蛋白相对表达量和MMP-9、IL-8、VEGF水平均显著降低(P均<0.05)。结论非布索坦灌胃可改善百草枯对大鼠的肺损伤,其机制可能与抑制XO和相关的氧化应激、下调PI3K/Akt通路蛋白表达、抑制β-catenin蛋白表达有关。

Objective To observe the effect of intragastric administration of febuxostat on paraquat-induced lung in⁃jury in rats and to explore its mechanism.Methods Twenty-one rats were randomly divided into three groups:the con⁃trol group,paraquat group,and paraquat+febuxostat group.The rats in the paraquat group and paraquat+febuxostat group were given paraquat by gavage;in the paraquat+febuxostat group,febuxostat was given by gavage for 24 h,but rats in the control group and paraquat group were given the same volume of normal saline every day.Rats in each group were sacri⁃ficed after the intragastric administration for 14 d,the lung tissues were separated,and blood were taken from the heart.HE staining was used to observe the pathological changes of the lung tissues of rats in each group.The colorimetric method was used to detect the serum lactate dehydrogenase(LDH),and ELISA was used to detect the serum soluble receptor for glycation end products(sRAGE).Colorimetric method was used to detect oxidative stress markers xanthine oxidase(XO),superoxide dismutase(SOD),total antioxidant capacity(TAC),malondialdehyde(MDA),and hydroxyproline in the lung tissues.Western blotting was used to detect PI3K,Akt,β-catenin protein in rat lung tissues,and double anti body sandwich ELISA method was used to detect matrix metalloproteinase-9(MMP-9),interleukin-8(IL-8),and vascu⁃lar endothelial growth factor(VEGF)in the lung tissues.Results The alveolar structure of rats was normal in the control group,but severe fibrous tissue hyperplasia,mixed inflammatory cell infiltration and pulmonary edema were found in rats of the paraquat group,snd the lung tissues of rats in the paraquat+febuxostat group showed moderate fibrous tissue hyper⁃plasia and inflammatory cell infiltration.Compared with the control group,the serum LDH level and sRAGE level signifi⁃cantly increased in the paraquat group(both P<0.05).Compared with the paraquat group,the LDH level in the paraquat+febuxostat group decreased significantly,and sRAGE level increased(both P<0.05).Compared with the control group,the levels of XO,MDA,and hydroxyproline significantly increased in the paraquat group(all P<0.05),and the levels of SOD and TAC were significantly reduced(both P<0.05).Compare with the paraquat group,the XO,MDA,and hydroxy⁃proline levels were significantly reduced(all P<0.05),and SOD and TAC levels increased significantly in the paraquat+febuxostat group(both P<0.05).Compared with the control group,the relative expression of PI3K,Akt,β-catenin pro⁃tein and the levels of MMP-9,IL-8 and VEGF in the lung tissues of rats in the paraquat group significantly increased(all P<0.05).Compared with the paraquat group,the relative expression of PI3K,Akt,β-catenin protein and the levels of MMP-9,IL-8,and VEGF in the lung tissues of rats were significantly reduced in the paraquat-febuxostat group(all P<0.05).Conclusion Intragastric administration of febuxostat can alleviate the paraquat-induced lung injury in rats,and the mechanism may be related to the inhibition of XO and related oxidative stress,down-regulation of PI3K/Akt pathway protein expression,and inhibition ofβ-catenin protein expression.