中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2015年
2期
145-150
,共6页
万慧娟%苏红霞%吴玉瑛%赵玉林%周明辉
萬慧娟%囌紅霞%吳玉瑛%趙玉林%週明輝
만혜연%소홍하%오옥영%조옥림%주명휘
鼻炎,变应性%受体,胞质和核%半胱氨酸天冬氨酸蛋白酶1%白细胞介素1β%白细胞介素18%疾病模型,动物
鼻炎,變應性%受體,胞質和覈%半胱氨痠天鼕氨痠蛋白酶1%白細胞介素1β%白細胞介素18%疾病模型,動物
비염,변응성%수체,포질화핵%반광안산천동안산단백매1%백세포개소1β%백세포개소18%질병모형,동물
Rhinitis,allergic%Receptors,cytoplasmic and nuclear%Caspase 1%Interleukin-1 beta%Interleukin-18%Disease models,animal
目的 检测天然免疫因子NLRP3炎性小体及下游因子白细胞介素1β(IL-1β)/IL-18在变应性鼻炎(AR)大鼠模型中的表达情况及其意义.方法 40只健康SD大鼠,雌雄不限,随机数字表法分为对照组(A组)、AR模型1组(B组)、AR模型2组(C组)、AR模型3组(D组),每组10只.A组不加卵清蛋白,B、C、D组用卵清蛋白及氢氧化铝基础致敏后,用5％卵清蛋白生理盐水溶液分别滴鼻10、20、30 d,1次/d,激发完成记录大鼠鼻部症状并处死大鼠.苏木精-伊红(HE)染色观察嗜酸粒细胞(EOS)计数、免疫组化观察NLRP3、半胱氨酸天冬酶1(Caspase-1)在各组大鼠鼻黏膜的表达情况,酶联免疫吸附实验(ELISA)检测大鼠卵清蛋白特异性IgE (OVA-sIgE)、IL-18、IL-1β在各组大鼠外周血中的相对表达量及IL-1β、IL-18在各组大鼠鼻腔冲洗液中的含量.以SPSS 17.0软件进行统计学分析.结果 嗜酸粒细胞(EOS)计数、行为学评分、OVA-sIgE浓度在AR模型组高于对照组,差异有统计学意义(P值均＜0.05),AR模型组之间比较差异亦有统计学意义(P值均＜0.05).NLRP3在B、C、D组鼻黏膜中的相对表达量分别为48.80±10.75、71.80±16.98、100.32±13.91,高于对照组的17.47±5.59,差异有统计学意义(F=78.399,P＜0.05);Caspase-1在B、C、D组鼻黏膜中的相对表达量分别为36.33 ±4.71、50.87±11.18、73.10±14.77,高于对照组的11.48±2.70,差异有统计学意义(F=71.727,P＜0.05);IL-1β在B、C、D组外周血中的含量分别为(56.46±10.13)、(82.37±11.93)、(112.01±22.91) pg/ml,高于对照组的(38.26±4.66)pg/ml,差异有统计学意义(F =51.981,P＜0.05);IL-18在B、C、D组外周血中的含量分别为(177.92±23.63)、(194.33±20.78)、(234.06±31.70) pg/ml,高于对照组的(89.71±5.56)pg/ml,差异有统计学意义(F =73.295,P＜0.05).且AR模型组之间比较差异亦有统计学意义(P值均＜0.05).NLRP3表达量与行为学评分、OVA-sIgE浓度、EOS细胞计数呈正相关(r值分别为0.833、0.873、0.868,P值均＜0.01).结论 NLRP3炎性小体及下游因子IL-1β/IL-18参与了AR的发病过程,可能与AR的炎性反应程度有关.
目的 檢測天然免疫因子NLRP3炎性小體及下遊因子白細胞介素1β(IL-1β)/IL-18在變應性鼻炎(AR)大鼠模型中的錶達情況及其意義.方法 40隻健康SD大鼠,雌雄不限,隨機數字錶法分為對照組(A組)、AR模型1組(B組)、AR模型2組(C組)、AR模型3組(D組),每組10隻.A組不加卵清蛋白,B、C、D組用卵清蛋白及氫氧化鋁基礎緻敏後,用5％卵清蛋白生理鹽水溶液分彆滴鼻10、20、30 d,1次/d,激髮完成記錄大鼠鼻部癥狀併處死大鼠.囌木精-伊紅(HE)染色觀察嗜痠粒細胞(EOS)計數、免疫組化觀察NLRP3、半胱氨痠天鼕酶1(Caspase-1)在各組大鼠鼻黏膜的錶達情況,酶聯免疫吸附實驗(ELISA)檢測大鼠卵清蛋白特異性IgE (OVA-sIgE)、IL-18、IL-1β在各組大鼠外週血中的相對錶達量及IL-1β、IL-18在各組大鼠鼻腔遲洗液中的含量.以SPSS 17.0軟件進行統計學分析.結果 嗜痠粒細胞(EOS)計數、行為學評分、OVA-sIgE濃度在AR模型組高于對照組,差異有統計學意義(P值均＜0.05),AR模型組之間比較差異亦有統計學意義(P值均＜0.05).NLRP3在B、C、D組鼻黏膜中的相對錶達量分彆為48.80±10.75、71.80±16.98、100.32±13.91,高于對照組的17.47±5.59,差異有統計學意義(F=78.399,P＜0.05);Caspase-1在B、C、D組鼻黏膜中的相對錶達量分彆為36.33 ±4.71、50.87±11.18、73.10±14.77,高于對照組的11.48±2.70,差異有統計學意義(F=71.727,P＜0.05);IL-1β在B、C、D組外週血中的含量分彆為(56.46±10.13)、(82.37±11.93)、(112.01±22.91) pg/ml,高于對照組的(38.26±4.66)pg/ml,差異有統計學意義(F =51.981,P＜0.05);IL-18在B、C、D組外週血中的含量分彆為(177.92±23.63)、(194.33±20.78)、(234.06±31.70) pg/ml,高于對照組的(89.71±5.56)pg/ml,差異有統計學意義(F =73.295,P＜0.05).且AR模型組之間比較差異亦有統計學意義(P值均＜0.05).NLRP3錶達量與行為學評分、OVA-sIgE濃度、EOS細胞計數呈正相關(r值分彆為0.833、0.873、0.868,P值均＜0.01).結論 NLRP3炎性小體及下遊因子IL-1β/IL-18參與瞭AR的髮病過程,可能與AR的炎性反應程度有關.
목적 검측천연면역인자NLRP3염성소체급하유인자백세포개소1β(IL-1β)/IL-18재변응성비염(AR)대서모형중적표체정황급기의의.방법 40지건강SD대서,자웅불한,수궤수자표법분위대조조(A조)、AR모형1조(B조)、AR모형2조(C조)、AR모형3조(D조),매조10지.A조불가란청단백,B、C、D조용란청단백급경양화려기출치민후,용5％란청단백생리염수용액분별적비10、20、30 d,1차/d,격발완성기록대서비부증상병처사대서.소목정-이홍(HE)염색관찰기산립세포(EOS)계수、면역조화관찰NLRP3、반광안산천동매1(Caspase-1)재각조대서비점막적표체정황,매련면역흡부실험(ELISA)검측대서란청단백특이성IgE (OVA-sIgE)、IL-18、IL-1β재각조대서외주혈중적상대표체량급IL-1β、IL-18재각조대서비강충세액중적함량.이SPSS 17.0연건진행통계학분석.결과 기산립세포(EOS)계수、행위학평분、OVA-sIgE농도재AR모형조고우대조조,차이유통계학의의(P치균＜0.05),AR모형조지간비교차이역유통계학의의(P치균＜0.05).NLRP3재B、C、D조비점막중적상대표체량분별위48.80±10.75、71.80±16.98、100.32±13.91,고우대조조적17.47±5.59,차이유통계학의의(F=78.399,P＜0.05);Caspase-1재B、C、D조비점막중적상대표체량분별위36.33 ±4.71、50.87±11.18、73.10±14.77,고우대조조적11.48±2.70,차이유통계학의의(F=71.727,P＜0.05);IL-1β재B、C、D조외주혈중적함량분별위(56.46±10.13)、(82.37±11.93)、(112.01±22.91) pg/ml,고우대조조적(38.26±4.66)pg/ml,차이유통계학의의(F =51.981,P＜0.05);IL-18재B、C、D조외주혈중적함량분별위(177.92±23.63)、(194.33±20.78)、(234.06±31.70) pg/ml,고우대조조적(89.71±5.56)pg/ml,차이유통계학의의(F =73.295,P＜0.05).차AR모형조지간비교차이역유통계학의의(P치균＜0.05).NLRP3표체량여행위학평분、OVA-sIgE농도、EOS세포계수정정상관(r치분별위0.833、0.873、0.868,P치균＜0.01).결론 NLRP3염성소체급하유인자IL-1β/IL-18삼여료AR적발병과정,가능여AR적염성반응정도유관.
Objective To detect the expression and explore the role of the innate immune NLRP3 inflammasome and its downstream factors interleukin-1 β (IL-1β)/ interleukin-18 (IL-18) in rat model of allergic rhinitis (AR).Methods Forty Sprague Dawley (SD) rats were randomly divided into control group (A group),AR model group 1 (B group),AR model group 2 (C group),AR model group 3 (D group).Every group contained 10 rats.After the rats in the model group were sensitized by ovalbumin (OVA) and alum,B,C and D groups were separately stimulated with 5％ OVA for 10 days,20 days and 30 days (once a day).The control group did not add OVA in the process of sensitization and excitation.All rats were executed after excitation.Eosinophil granulocyte (EOS) infiltration were observed in nasal mucosa by hematoxylin-eosin (HE) staining,the expression of NLRP3 and cysteinyl aspartate-specific protease-1 (Caspase-1) were observed in nasal mucosa by immunohistochemical staining.The concentrations of ovalbumin specific IgE (OVA-sIgE),IL-18 and IL-1 β in peripheral blood and the concentrations of IL-18 and I L-1 β in nasal fluid were tested by enzyme-linked immunosorbent assay (ELISA).The data were processed by SPSS 17.0 software.Results EOS cell counted,the behavioral score and the concentrations of OVA-sIgE in AR model group were obviously higher than those in control group (P ＜ 0.05),and the difference of which had statistical significance between the AR model groups (P ＜ 0.05).The expression of NLRP3 in AR model group (The expression of NLRP3 in group of B,C and D were 48.80 ± 10.75,71.80 ± 16.98 and 100.32 ± 13.91,respectively)were obviously higher than those in control group (17.47 ± 5.59),the difference of which had statistical significance (F =78.399,P ＜0.05).The expression of Caspase-1 in AR model group (The expression of Caspase-1 in group of B,C and D were 36.33 ± 4.71,50.87 ± 11.18 and 73.10 ± 14.77,respectively) were obviously higher than those in control group (11.48 ±2.70),the difference of which had statistical significance (F =71.727,P ＜0.05).The concentrations of IL-1β in AR model group [The concentrations of IL-1β in group of B,C and D were (56.46 ± 10.13),(82.37 ± 11.93),(112.01 ± 22.91) pg/ml,respectively] were obviously higher than those in control group [(38.26 ±4.66) pg/ml],the difference of which had statistical significance (F =51.981,P ＜0.05).The concentrations of IL-18 in AR model group [The concentrations of IL-18 in group of B,C and D were (177.92 ± 23.63),(194.33 ± 20.78),(234.06 ± 31.70) pg/ml,respectively] were obviously higher than those in control group [(89.71 ± 5.56) pg/ml],the difference of which had statistical significance (F =73.295,P ＜ 0.05).And the difference of which had statistical significance between the AR model groups (P ＜ 0.05).The expression of NLRP3 was significantly positively correlated with the behavioral score,the concentrations of OVA-sIgE and EOS cell counted in rat model of allergic rhinitis (r value were 0.833,0.873 and 0.868,respectively,all P ＜0.01).Conclusion NLRP3 inflammasome and its downstream factors IL-1β/IL-18 play a role in the pathogenesis of allergic rhinitis,which may be correlated with the degree of inflammation.
