中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2012年
7期
497-502
,共6页
李洪涛%张天托%陈壮桂%黄静%刘慧%朱家馨
李洪濤%張天託%陳壯桂%黃靜%劉慧%硃傢馨
리홍도%장천탁%진장계%황정%류혜%주가형
布地奈德%胸腺基质淋巴生成素%哮喘%树突细胞%细胞因子
佈地奈德%胸腺基質淋巴生成素%哮喘%樹突細胞%細胞因子
포지내덕%흉선기질림파생성소%효천%수돌세포%세포인자
Budesonide% Thymic Stromal Lymphopoetin% Asthma% Dendritic cells%Cytokine
目的 探讨布地奈德对支气管哮喘(简称哮喘)小鼠胸腺基质淋巴生成素(TSLP)-树突细胞(DC)途径和Th2极化的影响及其机制.方法 将18只清洁级雌性6～8周龄BALB/c小鼠按随机数字表法分为哮喘组[卵清白蛋白(OVA)腹腔致敏,OVA吸入激发]、布地奈德(BUD)干预组(OVA腹腔致敏,气道内吸入OVA和布地奈德)和对照组(以生理盐水代替OVA混悬液腹腔注射及雾化吸入),每组6只.观察各组小鼠生物学及肺组织病理学改变,收集BALF,并分离培养小鼠脾脏树突细胞,酶联免疫吸附试验( ELISA)检测BALF中TSLP水平、树突细胞上清液中TSLP受体(TSLPR)水平和DC-T细胞共培养上清液中细胞因子.流式细胞仪检测树突细胞表型.多组间比较采用单因素方差分析,组间两两比较采用t检验.结果 布地奈德干预组支气管及血管周围炎症细胞浸润轻于哮喘组.哮喘组BALF中TSLP水平[(44.0±5.1)ng/L]和树突细胞培养上清液中TSLPR水平[(19.7±2.2) ng/L]均高于对照组[分别为(14.2±3.6) ng/L和(10.4±1.2) ng/L,t值分别为11.74和9.13,均P＜0.01],BALF中TSLP水平与IL-5水平和嗜酸粒细胞呈正相关(r值分别为0.89和0.82,均P＜0.05)；布地奈德干预组BALF中TSLP水平[(19.2±5.8) ng/L]及树突细胞表达TSLPR水平[(12.5 ±2.8) ng/L]均低于哮喘组[分别为(44.0±5.1)ng/L和(19.7±2.2)ng/L],差异有统计学意义(t值分别为-7.86和-4.97,均P＜0.01)；布地奈德干预组BALF及DC-T细胞共培养上清液中IL-5[分别为(41±4)ng/L和(28 ±9) ng/L]低于哮喘组[分别为(81 ±8) ng/L和(51±14) ng/L],差异有统计学意义(t值分别为-11.00和-3.35,均P＜0.01),3组间IFN-γ差异无统计学意义(F值分别为0.97和0.82,均P＞0.05)；布地奈德干预组与哮喘组相比CD40、CD80和CD86等树突细胞表型下降.结论 布地奈德影响树突细胞表型,下调BALF中TSLP水平和树突细胞表达TSLPR水平,可能通过影响TSLP-DC途径而抑制哮喘Th2极化反应.
目的 探討佈地奈德對支氣管哮喘(簡稱哮喘)小鼠胸腺基質淋巴生成素(TSLP)-樹突細胞(DC)途徑和Th2極化的影響及其機製.方法 將18隻清潔級雌性6～8週齡BALB/c小鼠按隨機數字錶法分為哮喘組[卵清白蛋白(OVA)腹腔緻敏,OVA吸入激髮]、佈地奈德(BUD)榦預組(OVA腹腔緻敏,氣道內吸入OVA和佈地奈德)和對照組(以生理鹽水代替OVA混懸液腹腔註射及霧化吸入),每組6隻.觀察各組小鼠生物學及肺組織病理學改變,收集BALF,併分離培養小鼠脾髒樹突細胞,酶聯免疫吸附試驗( ELISA)檢測BALF中TSLP水平、樹突細胞上清液中TSLP受體(TSLPR)水平和DC-T細胞共培養上清液中細胞因子.流式細胞儀檢測樹突細胞錶型.多組間比較採用單因素方差分析,組間兩兩比較採用t檢驗.結果 佈地奈德榦預組支氣管及血管週圍炎癥細胞浸潤輕于哮喘組.哮喘組BALF中TSLP水平[(44.0±5.1)ng/L]和樹突細胞培養上清液中TSLPR水平[(19.7±2.2) ng/L]均高于對照組[分彆為(14.2±3.6) ng/L和(10.4±1.2) ng/L,t值分彆為11.74和9.13,均P＜0.01],BALF中TSLP水平與IL-5水平和嗜痠粒細胞呈正相關(r值分彆為0.89和0.82,均P＜0.05)；佈地奈德榦預組BALF中TSLP水平[(19.2±5.8) ng/L]及樹突細胞錶達TSLPR水平[(12.5 ±2.8) ng/L]均低于哮喘組[分彆為(44.0±5.1)ng/L和(19.7±2.2)ng/L],差異有統計學意義(t值分彆為-7.86和-4.97,均P＜0.01)；佈地奈德榦預組BALF及DC-T細胞共培養上清液中IL-5[分彆為(41±4)ng/L和(28 ±9) ng/L]低于哮喘組[分彆為(81 ±8) ng/L和(51±14) ng/L],差異有統計學意義(t值分彆為-11.00和-3.35,均P＜0.01),3組間IFN-γ差異無統計學意義(F值分彆為0.97和0.82,均P＞0.05)；佈地奈德榦預組與哮喘組相比CD40、CD80和CD86等樹突細胞錶型下降.結論 佈地奈德影響樹突細胞錶型,下調BALF中TSLP水平和樹突細胞錶達TSLPR水平,可能通過影響TSLP-DC途徑而抑製哮喘Th2極化反應.
목적 탐토포지내덕대지기관효천(간칭효천)소서흉선기질림파생성소(TSLP)-수돌세포(DC)도경화Th2겁화적영향급기궤제.방법 장18지청길급자성6～8주령BALB/c소서안수궤수자표법분위효천조[란청백단백(OVA)복강치민,OVA흡입격발]、포지내덕(BUD)간예조(OVA복강치민,기도내흡입OVA화포지내덕)화대조조(이생리염수대체OVA혼현액복강주사급무화흡입),매조6지.관찰각조소서생물학급폐조직병이학개변,수집BALF,병분리배양소서비장수돌세포,매련면역흡부시험( ELISA)검측BALF중TSLP수평、수돌세포상청액중TSLP수체(TSLPR)수평화DC-T세포공배양상청액중세포인자.류식세포의검측수돌세포표형.다조간비교채용단인소방차분석,조간량량비교채용t검험.결과 포지내덕간예조지기관급혈관주위염증세포침윤경우효천조.효천조BALF중TSLP수평[(44.0±5.1)ng/L]화수돌세포배양상청액중TSLPR수평[(19.7±2.2) ng/L]균고우대조조[분별위(14.2±3.6) ng/L화(10.4±1.2) ng/L,t치분별위11.74화9.13,균P＜0.01],BALF중TSLP수평여IL-5수평화기산립세포정정상관(r치분별위0.89화0.82,균P＜0.05)；포지내덕간예조BALF중TSLP수평[(19.2±5.8) ng/L]급수돌세포표체TSLPR수평[(12.5 ±2.8) ng/L]균저우효천조[분별위(44.0±5.1)ng/L화(19.7±2.2)ng/L],차이유통계학의의(t치분별위-7.86화-4.97,균P＜0.01)；포지내덕간예조BALF급DC-T세포공배양상청액중IL-5[분별위(41±4)ng/L화(28 ±9) ng/L]저우효천조[분별위(81 ±8) ng/L화(51±14) ng/L],차이유통계학의의(t치분별위-11.00화-3.35,균P＜0.01),3조간IFN-γ차이무통계학의의(F치분별위0.97화0.82,균P＞0.05)；포지내덕간예조여효천조상비CD40、CD80화CD86등수돌세포표형하강.결론 포지내덕영향수돌세포표형,하조BALF중TSLP수평화수돌세포표체TSLPR수평,가능통과영향TSLP-DC도경이억제효천Th2겁화반응.
Objective To investigate the effect of budesonide (BUD) on thymic stromal lymphopoetin receptor (TSLPR) of dendritic cells (DCs) in OVA-induced mouse asthma models and to explore the mechanisms by studying the effect of BUD on function of DCs from the model Methods Eighteen BALB/c female mice were randomly divided into a control group,an asthma group and a BUD intervention group,with 6 mice in each group.Mice were sensitized and challenged with ovalbumin (OVA) to establish the asthmatic model.The bronchoalveolar lavage fluid (BALF) and DCs of spleen from the 3 groups were harvested and the supematants of BALF and DCs were analyzed for levels of TSLP and TSLPR,respectively,by commercially available ELISA kit.The percentage of eosinophils ( EOS ) in BALF was counted.The expression of CD40,CD80 and CD86 in DCs was detected by FACS. The DCs were then washed,and co-cultured in vitro with autologous T cells purified by a nylon cotton column.The supernatants of DC-T co-culture were collected after 72 h incubation,and analyzed for levels of interleukin-5 ( IL-5 ) and interferon-γ (IFN-γ) by ELISA.Results The levels of TSLP in BALF and TSLPR in DCs from the asthma group were significantly increased compared with the control group [ (44.0 ± 5.1 ) ng/L vs ( 14.2 ± 3.6 )ng/L,P ＜0.01 and ( 19.7 ±2.2) ng/L vs ( 10.4 ± 1.2) ng/L,P ＜0.05,respectively].The expressionof CD40,CD80 and CD86 of DCs and IL-5 in the culture supernatants of DC-T co-culture was significant upregulated in the asthma group compared with the control group ( P ＜ 0.05 ).Furthermore,the addition of BUD reduced the expression of CD40,CD80,CD86,TSLPR in DCs,IL-5 in the culture supematants of DCT co-culture,TSLP and EOS in BALF.The level of INF-γ in the DC-T co-culture supernatants of the 3 groups did not achieve statistical significance ( F =0.82,P ＞ 0.05 ). Conclusion These results demonstrate that the therapeutic activity of BUD in asthmatic mice may be related to modulation of Th1 and Th2 cell functions and this effect is probably mediated through the TSLP-DC pathway.
