中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2013年
4期
281-286
,共6页
梁海梅%于化鹏%郑燕妮%夏虎%邓火金%樊慧珍%龚雨新%方泽葵%刁建新
樑海梅%于化鵬%鄭燕妮%夏虎%鄧火金%樊慧珍%龔雨新%方澤葵%刁建新
량해매%우화붕%정연니%하호%산화금%번혜진%공우신%방택규%조건신
NLRP3%IL-1β%IL-18%哮喘小鼠
NLRP3%IL-1β%IL-18%哮喘小鼠
NLRP3%IL-1β%IL-18%효천소서
NLRP3%IL-1β%IL-18%Allergic airway disease
目的 探讨NLRP3(nucleotide-binding oligomerization domain-leucine-rich repeats containing pyrin domain 3)/IL-1β、IL-18通路在哮喘小鼠肺支气管炎症的关系.方法 C57BL/6小鼠随机分为正常对照组、NLRP3抑制剂格列本脲组和哮喘组,每组7只；格列本脲组和哮喘组采用卵蛋白致敏、诱导建立哮喘小鼠模型,格列本脲组每次雾化激发前30 min腹腔注射格列本脲500 mg/kg;造模后检测小鼠气道反应性、苏木精-伊红(HE)染色观察小鼠肺组织病理改变、计数支气管肺泡灌洗液(BALF)中白细胞总数及嗜酸细胞百分比、淋巴细胞百分比以了解各组小鼠的支气管炎症程度；采用Western blot和RT-PCR检测小鼠肺组织NLRP3蛋白和mRNA的表达；酶联免疫吸附试验(ELISA)检测小鼠肺组织匀浆上清IL-1β、IL-18含量；并分别对IL-1β、IL-18与BALF中嗜酸性粒细胞百分比进行相关性分析.结果 哮喘组、格列本脲组气道反应性均高于对照组(P＜0.05).哮喘组BALF中白细胞总数、嗜酸粒细胞百分比及淋巴细胞百分比[(29.88±4.97) ×104/ml、(21.67±4.83)％、(31.87±5.31)％]均明显高于格列本脲组[(17.30±1.19) ×104/ml、(12.45±1.12)％、(17.16±0.97)％,P均＜0.05]及对照组[(9.74 ±2.88)×104/ml、(1.15±0.26)％、(10.66±1.83)％,P均＜0.05],而格列本脲组比对照组高(P均＜0.05).哮喘组、格列本脲组肺组织病理检查可见支气管周围较多炎症细胞浸润,但格列本脲组较哮喘组炎症浸润少,而对照组肺组织基本无炎症细胞浸润.哮喘组肺组织NLRP3蛋白及mRNA表达均高于格列本脲组、对照组(P均＜0.05).哮喘组肺组织匀浆上清IL-1β(74.81±17.45) pg/mg及IL-18(426.94±76.05) pg/mg含量均高于对照组[(37.54±5.53) pg/mg,P＜0.05；(249.62±161.20) pg/mg,P＜0.05].IL-1β、IL-18与嗜酸性粒细胞百分比正相关(r=0.833,P=0.02; r=0.856,P=0.014).结论 NLRP3及其下游因子IL-1β、IL-18在哮喘小鼠肺组织中高表达,采用格列本脲进行干预后哮喘小鼠支气管炎症减轻,提示NLRP3/IL-1β、IL-18通路可能在支气管-炎症的形成中发挥着重要作用.
目的 探討NLRP3(nucleotide-binding oligomerization domain-leucine-rich repeats containing pyrin domain 3)/IL-1β、IL-18通路在哮喘小鼠肺支氣管炎癥的關繫.方法 C57BL/6小鼠隨機分為正常對照組、NLRP3抑製劑格列本脲組和哮喘組,每組7隻；格列本脲組和哮喘組採用卵蛋白緻敏、誘導建立哮喘小鼠模型,格列本脲組每次霧化激髮前30 min腹腔註射格列本脲500 mg/kg;造模後檢測小鼠氣道反應性、囌木精-伊紅(HE)染色觀察小鼠肺組織病理改變、計數支氣管肺泡灌洗液(BALF)中白細胞總數及嗜痠細胞百分比、淋巴細胞百分比以瞭解各組小鼠的支氣管炎癥程度；採用Western blot和RT-PCR檢測小鼠肺組織NLRP3蛋白和mRNA的錶達；酶聯免疫吸附試驗(ELISA)檢測小鼠肺組織勻漿上清IL-1β、IL-18含量；併分彆對IL-1β、IL-18與BALF中嗜痠性粒細胞百分比進行相關性分析.結果 哮喘組、格列本脲組氣道反應性均高于對照組(P＜0.05).哮喘組BALF中白細胞總數、嗜痠粒細胞百分比及淋巴細胞百分比[(29.88±4.97) ×104/ml、(21.67±4.83)％、(31.87±5.31)％]均明顯高于格列本脲組[(17.30±1.19) ×104/ml、(12.45±1.12)％、(17.16±0.97)％,P均＜0.05]及對照組[(9.74 ±2.88)×104/ml、(1.15±0.26)％、(10.66±1.83)％,P均＜0.05],而格列本脲組比對照組高(P均＜0.05).哮喘組、格列本脲組肺組織病理檢查可見支氣管週圍較多炎癥細胞浸潤,但格列本脲組較哮喘組炎癥浸潤少,而對照組肺組織基本無炎癥細胞浸潤.哮喘組肺組織NLRP3蛋白及mRNA錶達均高于格列本脲組、對照組(P均＜0.05).哮喘組肺組織勻漿上清IL-1β(74.81±17.45) pg/mg及IL-18(426.94±76.05) pg/mg含量均高于對照組[(37.54±5.53) pg/mg,P＜0.05；(249.62±161.20) pg/mg,P＜0.05].IL-1β、IL-18與嗜痠性粒細胞百分比正相關(r=0.833,P=0.02; r=0.856,P=0.014).結論 NLRP3及其下遊因子IL-1β、IL-18在哮喘小鼠肺組織中高錶達,採用格列本脲進行榦預後哮喘小鼠支氣管炎癥減輕,提示NLRP3/IL-1β、IL-18通路可能在支氣管-炎癥的形成中髮揮著重要作用.
목적 탐토NLRP3(nucleotide-binding oligomerization domain-leucine-rich repeats containing pyrin domain 3)/IL-1β、IL-18통로재효천소서폐지기관염증적관계.방법 C57BL/6소서수궤분위정상대조조、NLRP3억제제격렬본뇨조화효천조,매조7지；격렬본뇨조화효천조채용란단백치민、유도건립효천소서모형,격렬본뇨조매차무화격발전30 min복강주사격렬본뇨500 mg/kg;조모후검측소서기도반응성、소목정-이홍(HE)염색관찰소서폐조직병리개변、계수지기관폐포관세액(BALF)중백세포총수급기산세포백분비、림파세포백분비이료해각조소서적지기관염증정도；채용Western blot화RT-PCR검측소서폐조직NLRP3단백화mRNA적표체；매련면역흡부시험(ELISA)검측소서폐조직균장상청IL-1β、IL-18함량；병분별대IL-1β、IL-18여BALF중기산성립세포백분비진행상관성분석.결과 효천조、격렬본뇨조기도반응성균고우대조조(P＜0.05).효천조BALF중백세포총수、기산립세포백분비급림파세포백분비[(29.88±4.97) ×104/ml、(21.67±4.83)％、(31.87±5.31)％]균명현고우격렬본뇨조[(17.30±1.19) ×104/ml、(12.45±1.12)％、(17.16±0.97)％,P균＜0.05]급대조조[(9.74 ±2.88)×104/ml、(1.15±0.26)％、(10.66±1.83)％,P균＜0.05],이격렬본뇨조비대조조고(P균＜0.05).효천조、격렬본뇨조폐조직병리검사가견지기관주위교다염증세포침윤,단격렬본뇨조교효천조염증침윤소,이대조조폐조직기본무염증세포침윤.효천조폐조직NLRP3단백급mRNA표체균고우격렬본뇨조、대조조(P균＜0.05).효천조폐조직균장상청IL-1β(74.81±17.45) pg/mg급IL-18(426.94±76.05) pg/mg함량균고우대조조[(37.54±5.53) pg/mg,P＜0.05；(249.62±161.20) pg/mg,P＜0.05].IL-1β、IL-18여기산성립세포백분비정상관(r=0.833,P=0.02; r=0.856,P=0.014).결론 NLRP3급기하유인자IL-1β、IL-18재효천소서폐조직중고표체,채용격렬본뇨진행간예후효천소서지기관염증감경,제시NLRP3/IL-1β、IL-18통로가능재지기관-염증적형성중발휘착중요작용.
Objective To verify the hypothesis that NLRP3-dependent IL-1β and IL-18 play a pathogenic role in mice with asthma.Methods A total of 21 C57BL/6 mice were randomly divided into 3 groups:the control group,NLRP3 inhibitor glyburine group and asthma group (n =7).A mouse model of allergic asthma was established by sensitization and challenge with ovalbumin for the mice in glyburine group and asthma group.Glyburine at 0.5 mg/g was given to the mice in glyburine group by intraperitoneal injection 30 minutes before inhaling ovalbumin.After induction of asthma,the airway responsiveness was measured and the pathological change in lung tissues was observed with H&E staining.The counts of total leukocytes and the percentages of eosinophils and lymphocytes in bronchoalveolar lavage fluid (BALF) were calculated for evaluation of airway inflammation.The expressions of NLRP3 and its mRNA were identified by Western blot and RT-PCR,respectively.The concentrations of IL-1β and IL-18 in mouse lung homogenate supernatant were detected by ELISA.Then the correlations between IL-1β/IL-18 and the percentage of eosinophils in BALF was analyzed.Results The airway responsiveness of the mice in asthma and glyburine groups were significantly higher than those in control group (P＜0.05).The total cell numbers of leukocytes and the percentages of eosinophils and lymphocytes in BALF of the mice in asthma group were the highest among three groups (P＜0.05),followed by the glyburine group (P＜0.05).The airway inflammation could be detected in both group B and C,however,be alleviated by glyburine intervention.The expressions of NL-RP3 protein and mRNA in the mice with asthma were higher than those in the control group and the glyburine group (P＜0.05).The concentrations of IL-1β and IL-18 were increased in the mice in asthma group as compared with those in the control group.The correlative analysis showed that there was a positive correlation between IL-1β/IL-18 and the percentage of eosinophils (r=0.833,P=0.02; r=0.856,P=0.014).Conclusion High expressions of NLRP3,IL-1β and IL-18 in the lungs of mice with asthma could be inhibited with glyburine treatment,suggesting NLRP3-dependent IL-1β and IL-18 play a crucial role in the development of allergic airway disease.
