摘要
目的:明确炎调方对于脂多糖(LPS)所致急性肺损伤(ALI)的抗炎和肺保护作用,探究炎调方在体内对花生四烯酸(AA)类代谢途径的影响.方法:30只雄性C57BL/6J小鼠按体质量随机分为正常组、模型组、地塞米松组(2mg·kg-1)、炎调方低剂量组(18g·kg-1)、炎调方高剂量组(36g·kg-1),每组6只.采用腹腔注射LPS建立小鼠ALI模型,给药组每天灌胃1次,连续灌胃7 d,实验结束时收集血清和肺组织.采用酶联免疫吸附测定法(ELISA)检测血清促炎因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)的含量;苏木素-伊红(HE)染色评估肺组织病理学变化;测定肺组织湿/干质量比(W/D);测定肺组织髓过氧化物酶(MPO)活力;采用液质联用技术检测血清与肺组织中AA类代谢物含量.结果:与正常组比较,模型组血清促炎因子TNF-α、IL-1β、IL-6的含量显著升高(P＜0.01);小鼠肺泡结构显著破坏,肺泡壁显著增厚,并伴有大量炎症细胞浸润;肺组织W/D和MPO活力均显著升高(P＜0.01);血清和肺组织中AA类代谢物前列腺素D2(PGD2)、前列腺素E2(PGE2)、11(S)-羟基二十碳四烯酸[11(S)-HETE]、5-羟基二十碳四烯酸(5-HETE)的含量明显升高(P＜0.05),血清11,12-环氧二十碳三烯酸(11,12-EET)、14,15-环氧二十碳三烯酸(14,15-EET)的含量显著降低(P＜0.01).与模型组比较,地塞米松组、炎调方低剂量组、炎调方高剂量组血清促炎因子TNF-α、IL-1β、IL-6的含量均明显降低(P＜0.05);小鼠肺泡壁轻度增厚,散有炎症细胞浸润,组织结构相对完整,肺泡结构有所改善;肺组织W/D和MPO活力均显著降低(P＜0.01);地塞米松组血清中AA类代谢物PGD2、PGE2、11(S)-HETE、5-HETE的含量明显降低(P＜0.05),血清14,15-EET的含量显著升高(P＜0.01),肺组织中5-HETE的含量显著降低(P＜0.01),炎调方低剂量组、炎调方高剂量组血清和肺组织中AA类代谢物PGE2、11(S)-HETE、5-HETE的含量均明显降低(P＜0.05),血清和肺组织中11,12-EET的含量均明显升高(P＜0.05).结论:炎调方对LPS诱导的ALI具有显著的保护作用,该作用与其调节体内AA的代谢途径有关.
Abstract
Objective:To clarify the anti-inflammatory and lung-protective effects of Yantiao prescription on lipopolysaccharide(LPS)-induced acute lung injury(ALI),and to explore the impact of Yantiao prescription on the metabolic pathways of arachidonic acid(AA)in vivo.Methods:Thirty male C57BL/6J mice were randomly divided into the following groups based on body weight:normal group,model group,dexamethasone group(2 mg·kg-1),low-dose Yantiao prescription group(18 g·kg-1),and high-dose Yantiao prescription group(36 g·kg-1),with 6 mice in each group.The ALI mouse model was established by intraperitoneal injection of LPS.The treatment groups received oral gavage once a day for 7 consecutive days,and serum and lung tissue were collected at the end of the experiment.The content of pro-inflammatory cytokines tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in serum was detected by enzyme-linked immunosorbent assay(ELISA).Hematoxylin-eosin(HE)staining was used to assess lung tissue pathology.The wet/dry weight ratio(W/D)and myeloperoxidase(MPO)activity in lung tissue were measured.The content of A A metabolites in serum and lung tissue was measured by liquid chromatography triple quadrupole-mass spectrometry(LC-MS/MS).Results:Compared with the conditions in the normal group,the content of serum pro-inflammatory cytokines TNF-α,IL-1β,and IL-6 in the model group was significantly increased(P＜0.01).The alveolar structure in mice was severely damaged,with markedly thickened alveolar walls and extensive inflammatory cell infiltration.The W/D ratio and MPO activity in lung tissue were significantly increased(P＜0.01).The content of AA metabolites,including prostaglandin D2(PGD2),prostaglandin E2(PGE2),11(S)-hydroxy-eicosatetraenoic acid[11(S)-HETE],and 5-hydroxy-eicosatetraenoic acid(5-HETE)in serum and lung tissue was significantly increased(P＜0.05),while the content of 11,12-epoxyeicosatrienoic acid(11,12-EET)and 14,15-epoxyeicosatrienoic acid(14,15-EET)in serum was significantly decreased(P＜0.01).Compared with the results in the model group,the content of serum pro-inflammatory cytokines TNF-α,IL-1β,and IL-6 in the dexamethasone group,low-dose Yantiao prescription group,and high-dose Yantiao prescription group was significantly reduced(P＜0.05).Mild thickening of alveolar walls,scattered inflammatory cell infiltration,and relatively intact tissue structure with improved alveolar architecture were observed.The W/D ratio and MPO activity in lung tissue were significantly reduced(P＜0.01).The content of AA metabolites PGD2,PGE2,11(S)-HETE,and 5-HETE in serum from the dexamethasone group was significantly decreased(P＜0.05),while the content of 14,15-EET in serum significantly increased(P＜0.01),and the content of 5-HETE in lung tissue significantly decreased(P＜0.01).In the low-dose and high-dose Yantiao prescription groups,the content of AA metabolites PGD2,PGE2,11(S)-HETE,and 5-HETE in serum and lung tissue was significantly decreased(P＜0.05),while the content of 11,12-EET in both serum and lung tissue was significantly increased(P＜0.05).Conclusion:Yantiao prescription has significant protective effects against LPS-induced ALI,which are related to its regulation of AA metabolic pathways in vivo.
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