柠檬苦素对脂多糖致小鼠急性肺损伤的作用
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摘要
目的:探讨柠檬苦素(LM)对脂多糖(LPS)致小鼠急性肺损伤的影响。方法:40只ICR雄性小鼠,按体质量随机分为4组:正常组、模型组、地塞米松(Dex)给药组和LM给药组,每组10只。Dex和LM均在滴注LPS前2 h分别小鼠腹腔注射10 mg/kg给药,之后除正常组小鼠只给予腹腔注射等体积的生理盐水外,其余小鼠气管滴注致死量的5 mg/kg LPS。LPS注射6 h后,处死小鼠,解剖小鼠取肺脏,计算小鼠肺指数、湿干比值。采用苏木素-伊红(HE)染色观察肺组织形态。采用ELISA法检测小鼠血清中TNF-α、IL-1β和IL-6含量;比色法检测血清髓过氧化物酶(MPO)含量。实时定量PCR检测TNF-α、IL-1β和IL-6mRNA表达情况。Western blot检测TLR4和NF-κB p65蛋白表达。结果:与正常组比较,模型组小鼠肺指数和湿干比值明显升高,肺组织间质炎性细胞浸润,伴明显的肺泡充血、水肿,血清MPO、TNF-α、IL-1β和IL-6含量及其肺组织mRNA表达量均明显升高(P<0.01);与模型组比较,LM给药组小鼠肺指数和湿干比值明显降低,肺组织病理状态明显改善,血清TNF-α、IL-1β和IL-6水平及肺组织TNF-αmRNA、IL-1βmRNA、IL-6 mRNA、TLR4和NF-κB p65蛋白表达量均明显降低(P<0.01)。结论:LM可通过调控TLR4/NF-κB通路抑制炎症因子的表达从而改善LPS诱导的小鼠肺组织损伤。
AIM: To investigate the effects of limonin( LM) on lipopolysaccharides( LPS)-induced acute lung injury( ALI) in mice.METHODS: Forty ICR male mice were divided randomly by weight into 4 groups: normal group,model group, dexamethasone( Dex)-treated group and LM-treated group. Each group had ten mice. Dex and LM were intraperitoneally injected at the dose of10 mg/kg,respectively. Two hours after drug administration,except the mice in normal group treated with same vehicle,others were intracheally injected with LPS( 5 mg/kg). Six hours after LPS challenge,mice were sacrificed and lung tissues were obtained for calculating the pulmonary index and wet/dry mass( W/D) ratio. The sections were stained with hematoxylin and eosin( HE) for histological analysis. The levels of TNF-α,IL-1β,and IL-6 in serum were determined by ELISA. Myeloperoxidase( MPO) in serum were determined by chromatometry. The mRNA expressions of TNF-α,IL-1β and IL-6 were determined by RT-PCR while TLR4 and NF-κB p65 protein by Western blot analysis. RESULTS: Compared with normal group,mice in model group had obvious increase of lung indexesand W/D ratios, which were accompanied by marked inflammatory cell infiltration,alveolar congestion and edema in lung; the levels of TNF-α,IL-1β,IL-6 and MPO in serum as well as their mRNA expressions in the lung of ALI mice were also increased( P < 0. 01). Compared with model group,lung indexes and W/D ratios were decreased in LMtreated group while histological changes were improved; the levels of TNF-α,IL-1β,and IL-6 in serum( P < 0. 01) and their mRNA expressions,and the expressions of TLR4 and NF-κB p65 protein in lung tissues were also significantly decreased( P< 0. 01). CONCLUSION: LM protects against LPS-induced ALI via regulating TLR4/NF-κB pathway and inhibiting expression of inflammatory cytokines in mice.
AIM: To investigate the effects of limonin( LM) on lipopolysaccharides( LPS)-induced acute lung injury( ALI) in mice.METHODS: Forty ICR male mice were divided randomly by weight into 4 groups: normal group,model group, dexamethasone( Dex)-treated group and LM-treated group. Each group had ten mice. Dex and LM were intraperitoneally injected at the dose of10 mg/kg,respectively. Two hours after drug administration,except the mice in normal group treated with same vehicle,others were intracheally injected with LPS( 5 mg/kg). Six hours after LPS challenge,mice were sacrificed and lung tissues were obtained for calculating the pulmonary index and wet/dry mass( W/D) ratio. The sections were stained with hematoxylin and eosin( HE) for histological analysis. The levels of TNF-α,IL-1β,and IL-6 in serum were determined by ELISA. Myeloperoxidase( MPO) in serum were determined by chromatometry. The mRNA expressions of TNF-α,IL-1β and IL-6 were determined by RT-PCR while TLR4 and NF-κB p65 protein by Western blot analysis. RESULTS: Compared with normal group,mice in model group had obvious increase of lung indexesand W/D ratios, which were accompanied by marked inflammatory cell infiltration,alveolar congestion and edema in lung; the levels of TNF-α,IL-1β,IL-6 and MPO in serum as well as their mRNA expressions in the lung of ALI mice were also increased( P < 0. 01). Compared with model group,lung indexes and W/D ratios were decreased in LMtreated group while histological changes were improved; the levels of TNF-α,IL-1β,and IL-6 in serum( P < 0. 01) and their mRNA expressions,and the expressions of TLR4 and NF-κB p65 protein in lung tissues were also significantly decreased( P< 0. 01). CONCLUSION: LM protects against LPS-induced ALI via regulating TLR4/NF-κB pathway and inhibiting expression of inflammatory cytokines in mice.
引文
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[11]郭长满,陈丹,宋东,等.右美托咪定对小鼠肺血-再灌注损伤的保护作用[J].中国临床药理学与治疗学,2016,21(2):145-149.
[12]Akbarshahi H,Rosendahl AH,Westergren-Thorsson G,et al.Acute lung injury in acute pancreatitis-Awaiting the big leap[J].Res Med,2012,106:1199-1210.
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[14]Samanta S,Rajasingh S,Cao T,et al.Epigenetic dysfunctional diseases and therapy for infection and inflammation[J].Biochim Biophys Acta,2017,1863(2):518-528.
[15]张利鹏,赵焱,刘国娟,等.乌司他丁通过干预p38MAPK/ERK信号通路减轻脓毒症性肺损伤[J].中国药理学通报,2016,32(9):1311-1316.
[16]钟海潮,范芳华.IL-32在重症肺炎患者血浆中表达的临床意义及其机制探讨[J].中国临床药理学与治疗学,2016,21(9):1024-1027.
[17]王檀,袁成波.温肺通痹颗粒治疗间质性肺疾病机理研究[J].长春中医药大学学报,2015,31(3):547-548.
[18]常秀娟,张帅,江益平,等.从细胞因子风暴探讨热毒宁注射液抗大鼠急性肺损伤作用机制[J].中草药,2015,46(2):236-239.
[2]Liu H,Yu X,Yu S,et al.Molecular mechanisms in lipopolysaccharide-induced pulmonary endothelial barrier dysfunction[J].Int Immunopharmacol,2015,29(2):937-946.
[3]冯燕妮,潘红飞,梁玉美.呼吸窘迫综合征呼吸机相关性肺损伤应用肺保护性通气策略的疗效分析[J].中国医药导报,2013,10(18):57-59.
[4]张红梅,王长虹,王峥涛.基于大鼠体内吸收分布的吴茱萸指标成分选择及测定[J].中成药,2016,38(7):1538-1544.
[5]张栋健,李薇,何庆文,等.UHPLC-Q-TOF-MS分析枳壳炮制前后成分变化[J].中国中药杂志,2016,41(11):2070-2080.
[6]Shimizu S,Miyamoto S,Fujii G,et al.Suppression of intestinal carcinogenesis in Apc-mutant mice by limonin[J].J Clin Biochem Nutr,2015,57(1):39-43.
[7]Yang N,Wang J,Liu C,et al.Berberine and limonin suppress Ig E production by human B cells and peripheral blood mononuclear cells from food-allergic patients[J].Ann Allergy Asthma Immunol,2014,113(5):556-564.
[8]胡毅翔,蔡伟,张欢欢,等.基于肺部苦味受体抗哮喘天然活性成分筛选模型的建立与应用[J].中草药,2016,47(5):775-780.
[9]周晓霞,张建情,刘春晓,等.白花前胡有效成分PdIa对急性肺损伤的作用及机制研究[J].中国药理学通报,2016,32(8):1165-1170.
[10]刘衍伶,邓林强,胡龙华,等.耐碳青霉烯类肺炎克雷伯菌耐药机制及呼吸机相关性感染的流行病学分析[J].中国临床药理学与治疗学,2015,20(10):1137-1143.
[11]郭长满,陈丹,宋东,等.右美托咪定对小鼠肺血-再灌注损伤的保护作用[J].中国临床药理学与治疗学,2016,21(2):145-149.
[12]Akbarshahi H,Rosendahl AH,Westergren-Thorsson G,et al.Acute lung injury in acute pancreatitis-Awaiting the big leap[J].Res Med,2012,106:1199-1210.
[13]畅毅平,彭鹏.炎症细胞因子与急性呼吸窘迫综合征的研究进展[J].临床肺科杂志,2015,20(2):315-318.
[14]Samanta S,Rajasingh S,Cao T,et al.Epigenetic dysfunctional diseases and therapy for infection and inflammation[J].Biochim Biophys Acta,2017,1863(2):518-528.
[15]张利鹏,赵焱,刘国娟,等.乌司他丁通过干预p38MAPK/ERK信号通路减轻脓毒症性肺损伤[J].中国药理学通报,2016,32(9):1311-1316.
[16]钟海潮,范芳华.IL-32在重症肺炎患者血浆中表达的临床意义及其机制探讨[J].中国临床药理学与治疗学,2016,21(9):1024-1027.
[17]王檀,袁成波.温肺通痹颗粒治疗间质性肺疾病机理研究[J].长春中医药大学学报,2015,31(3):547-548.
[18]常秀娟,张帅,江益平,等.从细胞因子风暴探讨热毒宁注射液抗大鼠急性肺损伤作用机制[J].中草药,2015,46(2):236-239.