CAJ | 학술논문

目的探讨甘草酸对支气管哮喘(哮喘)小鼠气道炎症的影响机制.方法将70只BALB/c小鼠采用随机数字表法分为对照组、哮喘组和低、中、高浓度甘草酸组,每组14只.哮喘组、各甘草酸组分别给予卵清白蛋白(OVA)致敏和激发.每次雾化前30 min,甘草酸组分别给予甘草酸(25、50、100 mg/kg)腹腔注射,对照组以生理盐水代替OVA腹腔注射及雾化吸入.末次激发后收集支气管肺泡灌洗液(BALF),离心后计数细胞总数及白细胞分类数量.酶联免疫吸附(ELISA)法测定BALF上清液中白细胞介素(IL)-12p70、IL-10、IL-4和干扰素(IFN)-γ水平.肺组织HE染色.流式细胞术检测小鼠脾脏CD11c+树突细胞表面CD86、主要组织相容复合物(MHC)-Ⅱ、CD40、OX40配体(OX40L)的表达水平.结果哮喘组BALF中细胞总数、嗜酸粒细胞(Eos)计数均显著高于对照组[(124.3±39.7)×107/L、(26.3±17.3)×107/L比(55.3±22.8)×107/L、(0.6±0.4)× 107/L],IL-10、IL-4水平也均显著高于对照组[(49±12)、(169±29)比(34±4)、(89±37) ng/L,而IFN-γ水平、IFN-γ/IL-4比值均显著低于对照组[(122±56) ng/L、0.7±-0.4比(89±37)ng/L、2.9±0.8](均P＜0.05).哮喘组脾脏树突细胞上CD86、MHC-Ⅱ、CD40、OX40L表达均显著高于对照组[(38.4±15.7)％、(90.4±3.4)％、(25.4±10.2)％、(29.6±9.9)％比(18.8±4.4)％、(73.1±11.3)％、(3.8±2.2)％、(5.0±1.6)％](均P＜0.05).高浓度甘草酸组BALF中细胞总数及Eos计数[(62.1±21.7) ×107/L、(2.2±1.7)×107/L]均显著低于哮喘组,IL-12p70、IL-10、IFN-γ水平和IFN-γ/IL-4比值[(44±14)、(132±13)、(207±66) ng/L和1.8 ±0.6]均显著高于哮喘组,IL-4水平[(122±38) ng/L]显著低于哮喘组,脾脏树突细胞上MHC-Ⅱ、CD40和OX40L表达[(75.8±15.9)％、(11.1±5.9)％、(11.8±3.4)％]显著低于哮喘组(均P ＜0.05).哮喘组小鼠气道黏膜充血水肿,气道壁及管周有大量以Eos为主的炎症细胞浸润,各甘草酸组气道炎症反应较哮喘组轻,在中高浓度甘草酸组尤其明显.结论甘草酸可能通过影响树突细胞表型,抑制哮喘Th2极化反应,减轻哮喘气道炎症. 目的探討甘草痠對支氣管哮喘(哮喘)小鼠氣道炎癥的影響機製.方法將70隻BALB/c小鼠採用隨機數字錶法分為對照組、哮喘組和低、中、高濃度甘草痠組,每組14隻.哮喘組、各甘草痠組分彆給予卵清白蛋白(OVA)緻敏和激髮.每次霧化前30 min,甘草痠組分彆給予甘草痠(25、50、100 mg/kg)腹腔註射,對照組以生理鹽水代替OVA腹腔註射及霧化吸入.末次激髮後收集支氣管肺泡灌洗液(BALF),離心後計數細胞總數及白細胞分類數量.酶聯免疫吸附(ELISA)法測定BALF上清液中白細胞介素(IL)-12p70、IL-10、IL-4和榦擾素(IFN)-γ水平.肺組織HE染色.流式細胞術檢測小鼠脾髒CD11c+樹突細胞錶麵CD86、主要組織相容複閤物(MHC)-Ⅱ、CD40、OX40配體(OX40L)的錶達水平.結果哮喘組BALF中細胞總數、嗜痠粒細胞(Eos)計數均顯著高于對照組[(124.3±39.7)×107/L、(26.3±17.3)×107/L比(55.3±22.8)×107/L、(0.6±0.4)× 107/L],IL-10、IL-4水平也均顯著高于對照組[(49±12)、(169±29)比(34±4)、(89±37) ng/L,而IFN-γ水平、IFN-γ/IL-4比值均顯著低于對照組[(122±56) ng/L、0.7±-0.4比(89±37)ng/L、2.9±0.8](均P＜0.05).哮喘組脾髒樹突細胞上CD86、MHC-Ⅱ、CD40、OX40L錶達均顯著高于對照組[(38.4±15.7)％、(90.4±3.4)％、(25.4±10.2)％、(29.6±9.9)％比(18.8±4.4)％、(73.1±11.3)％、(3.8±2.2)％、(5.0±1.6)％](均P＜0.05).高濃度甘草痠組BALF中細胞總數及Eos計數[(62.1±21.7) ×107/L、(2.2±1.7)×107/L]均顯著低于哮喘組,IL-12p70、IL-10、IFN-γ水平和IFN-γ/IL-4比值[(44±14)、(132±13)、(207±66) ng/L和1.8 ±0.6]均顯著高于哮喘組,IL-4水平[(122±38) ng/L]顯著低于哮喘組,脾髒樹突細胞上MHC-Ⅱ、CD40和OX40L錶達[(75.8±15.9)％、(11.1±5.9)％、(11.8±3.4)％]顯著低于哮喘組(均P ＜0.05).哮喘組小鼠氣道黏膜充血水腫,氣道壁及管週有大量以Eos為主的炎癥細胞浸潤,各甘草痠組氣道炎癥反應較哮喘組輕,在中高濃度甘草痠組尤其明顯.結論甘草痠可能通過影響樹突細胞錶型,抑製哮喘Th2極化反應,減輕哮喘氣道炎癥.
목적 탐토감초산대지기관효천(효천)소서기도염증적영향궤제.방법 장70지BALB/c소서채용수궤수자표법분위대조조、효천조화저、중、고농도감초산조,매조14지.효천조、각감초산조분별급여란청백단백(OVA)치민화격발.매차무화전30 min,감초산조분별급여감초산(25、50、100 mg/kg)복강주사,대조조이생리염수대체OVA복강주사급무화흡입.말차격발후수집지기관폐포관세액(BALF),리심후계수세포총수급백세포분류수량.매련면역흡부(ELISA)법측정BALF상청액중백세포개소(IL)-12p70、IL-10、IL-4화간우소(IFN)-γ수평.폐조직HE염색.류식세포술검측소서비장CD11c+수돌세포표면CD86、주요조직상용복합물(MHC)-Ⅱ、CD40、OX40배체(OX40L)적표체수평.결과 효천조BALF중세포총수、기산립세포(Eos)계수균현저고우대조조[(124.3±39.7)×107/L、(26.3±17.3)×107/L비(55.3±22.8)×107/L、(0.6±0.4)× 107/L],IL-10、IL-4수평야균현저고우대조조[(49±12)、(169±29)비(34±4)、(89±37) ng/L,이IFN-γ수평、IFN-γ/IL-4비치균현저저우대조조[(122±56) ng/L、0.7±-0.4비(89±37)ng/L、2.9±0.8](균P＜0.05).효천조비장수돌세포상CD86、MHC-Ⅱ、CD40、OX40L표체균현저고우대조조[(38.4±15.7)％、(90.4±3.4)％、(25.4±10.2)％、(29.6±9.9)％비(18.8±4.4)％、(73.1±11.3)％、(3.8±2.2)％、(5.0±1.6)％](균P＜0.05).고농도감초산조BALF중세포총수급Eos계수[(62.1±21.7) ×107/L、(2.2±1.7)×107/L]균현저저우효천조,IL-12p70、IL-10、IFN-γ수평화IFN-γ/IL-4비치[(44±14)、(132±13)、(207±66) ng/L화1.8 ±0.6]균현저고우효천조,IL-4수평[(122±38) ng/L]현저저우효천조,비장수돌세포상MHC-Ⅱ、CD40화OX40L표체[(75.8±15.9)％、(11.1±5.9)％、(11.8±3.4)％]현저저우효천조(균P ＜0.05).효천조소서기도점막충혈수종,기도벽급관주유대량이Eos위주적염증세포침윤,각감초산조기도염증반응교효천조경,재중고농도감초산조우기명현.결론 감초산가능통과영향수돌세포표형,억제효천Th2겁화반응,감경효천기도염증.
Objective To explore the therapeutic effects and mechanism of glycyrrhizic acid (GA) on airway inflammation in a murine model of asthma.Methods A total of 70 female BALB/c mice were randomly divided into 5 groups (n =14 each) of control,asthmatic and three treatments with low,medium and high doses of GA.Mice in the asthmatic and the treatment groups were sensitized and challenged by ovalbumin (OVA).Mice of the treatment groups were injected intraperitoneally with GA (25,50,100 mg/kg) 30 min before each OVA challenge.The control mice received an aerosol inhalation of normal saline instead of OVA.Within 24 hours after the last OVA challenge,bronchoalveolar lavage fluid (BALF) was collected for counting total cells and eosinophils (Eos) in other 8 mice in each group.Interleukin (IL)-12p70,IL-10,IL-4 and interferon gamma (IFN-γ) were measured in BALF by enzyme-linked immunosorbent assay (ELISA).Histological studies of lung were conducted with hematoxylin and eosin staining (HE) and the expressions of CD86,major histocompatibility complex (MHC)-Ⅱ,CD40,OX40 ligand (OX40L) on CD1 1c + dendritic cells (DCs) in spleens were evaluated with flow cytometry (FCM).Results Compared with the control group,the number of total cells and eosinophils was higher in BALF in asthmatic group ((124.3 ±39.7) ×107/L,(26.3 ±17.2) × 107/L vs (55.3 ±22.8) × 107/L,(0.6 ± 0.4) × 107/L),the expressions of IL-10 and IL-4 increased ((49 ± 12,169 ±29) ng/L vs (34 ±4,89 ± 37) ng/L) and the levels of IFN-γand IFN-γ/IL-4 ratio decreased in BALF ((122 ±56) ng/L,(0.7 ± 0.4) vs (89 ± 37) ng/L,2.9 ± 0.8)),the expressions of CD86,MHC-Ⅱ,CD40,OX40L on CD1 1 c + DCs increased in spleens ((38.4 ± 15.7) ％,(90.4 ± 3.4) ％,(25.4 ± 10.2) ％,(29.6 ± 9.9) ％ vs (18.8±4.4)％,(73.1±11.3)％,(3.8 ±2.2)％,(5.0 ±1.6)％) (allP＜0.05).Compared with the asthmatic group,total cell counts and the eosinophil numbers significantly decreased by the treatment of GA at a dose of 100 mg/kg ((62.1 ±21.7) × 107/L,(2.2 ± 1.7) × 107/L),the levels of IL-12p70,IL-10,IFN-γand the ratio of IFN-γ/IL-4 increased ((44 ± 14,132 ± 13,208 ± 66) ng/L,(1.8 ± 0.6)) and the level IL-4 decreased (122 ±38) ng/L.The expressions of MHC-Ⅱ,CD40,OX40L on CD11c+ DCs decreased ((75.8 ± 15.9) ％,(11.1 ± 5.9) ％,(11.8 ± 3.4) ％)) (all P ＜ 0.05).The asthmatic mice induced an infiltration of inflammatory cells around airways and blood vessels.Administration of GA significantly reduced the infiltration of inflammatory cells in peribronchial areas compared with asthmatic mice especially at a dose of 50 mg/kg 100 mg/kg.Conclusion GA effectively ameliorates the airway inflammation of asthma via inhibiting the Th2 responses though modulating the expressions of CD86,MHC-Ⅱ,CD40,OX40L on CD11c+ DCs.