国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2012年
18期
1373-1377
,共5页
应延风%陈浩浩%金耀建%屠平光%胡野
應延風%陳浩浩%金耀建%屠平光%鬍野
응연풍%진호호%금요건%도평광%호야
气道平滑肌细胞%前列腺素E2%白介素%γ-干扰素%凋亡
氣道平滑肌細胞%前列腺素E2%白介素%γ-榦擾素%凋亡
기도평활기세포%전렬선소E2%백개소%γ-간우소%조망
Airway smooth muscle cells%Prostaglandin E2%Interleukin%Interferon-γ%Apoptosis
目的 前列腺素E2(prostaglandin E2,PGE2)通过调控气道平滑肌细胞(airway smooth muscle cells,ASMCs)的辽移、分泌、增殖功能,对哮喘气道重塑有关键性影响.研究PGE2对ASMCs凋广及其分泌白介索4(interleukin 4,IL -4)、白介素10(interleukin 10,IL-10)和γ-干扰索(interferon-γ,IFN -γ)的影响,对阐明PGE2通过调控ASMCs分泌Th1/Th2细胞因子对气道炎症发挥作用机制有重要意义.方法 用大鼠气道平滑肌细胞株与终浓度为10 -9～10-6的PGE2共培养24 h,分6组,共4个浓度梯度.用Annexin V EGFP/PI双染细胞凋亡检测试剂盒在流式细胞仪测定凋亡率,同时用IL-4、IL- 10和IFN-γ ELISA试剂盒在酶标仪检测其浓度.每4个复孔为1组,实验重复2次,数据为8孔的平均值.结果 应用不同浓度的PGE2后ASMCs各组细胞凋亡率与对照组相比无明显变化.但IL分泌与PGE2有显著的量效关系,IL-4和IL-10随PGE2浓度下降而分泌量显著下降,但IFN-γ随PGE2浓度下降而分泌量呈升高趋势,两者呈分离现象(空白对照、阴性对照、10-9 PGE2、10-8 PGE2、10 -7 PGE2、10 -6 PGE2组,IL- 4分别为6.77 ± 1.58、9.24±2.45、38.17±11.33、29.27±11.12、26.05±9.49、21.11±7.21;IL-10为4.8±1.06、6.35±2.11、71.89±12.03、43.68 ±11.25、28.89± 8.7与、24.31±6.67；IFN-γ为2.17±1.97、3.25±1.48、10.63±4.75、13.4±5.57、15.47±6.16、19.54±6.35).结论 PGE2能调控ASMCs分泌细胞因子,随着浓度梯度增加能促进Th1型细胞因子IFN-γ分泌,同时抑制Th2型细胞因子 IL-4、IL-10分泌,但对ASMCs凋亡无显善影响.认为在正常状态下 PGE2的分泌与维持气道内Th1/Th2的平衡有关.
目的 前列腺素E2(prostaglandin E2,PGE2)通過調控氣道平滑肌細胞(airway smooth muscle cells,ASMCs)的遼移、分泌、增殖功能,對哮喘氣道重塑有關鍵性影響.研究PGE2對ASMCs凋廣及其分泌白介索4(interleukin 4,IL -4)、白介素10(interleukin 10,IL-10)和γ-榦擾索(interferon-γ,IFN -γ)的影響,對闡明PGE2通過調控ASMCs分泌Th1/Th2細胞因子對氣道炎癥髮揮作用機製有重要意義.方法 用大鼠氣道平滑肌細胞株與終濃度為10 -9～10-6的PGE2共培養24 h,分6組,共4箇濃度梯度.用Annexin V EGFP/PI雙染細胞凋亡檢測試劑盒在流式細胞儀測定凋亡率,同時用IL-4、IL- 10和IFN-γ ELISA試劑盒在酶標儀檢測其濃度.每4箇複孔為1組,實驗重複2次,數據為8孔的平均值.結果 應用不同濃度的PGE2後ASMCs各組細胞凋亡率與對照組相比無明顯變化.但IL分泌與PGE2有顯著的量效關繫,IL-4和IL-10隨PGE2濃度下降而分泌量顯著下降,但IFN-γ隨PGE2濃度下降而分泌量呈升高趨勢,兩者呈分離現象(空白對照、陰性對照、10-9 PGE2、10-8 PGE2、10 -7 PGE2、10 -6 PGE2組,IL- 4分彆為6.77 ± 1.58、9.24±2.45、38.17±11.33、29.27±11.12、26.05±9.49、21.11±7.21;IL-10為4.8±1.06、6.35±2.11、71.89±12.03、43.68 ±11.25、28.89± 8.7與、24.31±6.67；IFN-γ為2.17±1.97、3.25±1.48、10.63±4.75、13.4±5.57、15.47±6.16、19.54±6.35).結論 PGE2能調控ASMCs分泌細胞因子,隨著濃度梯度增加能促進Th1型細胞因子IFN-γ分泌,同時抑製Th2型細胞因子 IL-4、IL-10分泌,但對ASMCs凋亡無顯善影響.認為在正常狀態下 PGE2的分泌與維持氣道內Th1/Th2的平衡有關.
목적 전렬선소E2(prostaglandin E2,PGE2)통과조공기도평활기세포(airway smooth muscle cells,ASMCs)적료이、분비、증식공능,대효천기도중소유관건성영향.연구PGE2대ASMCs조엄급기분비백개색4(interleukin 4,IL -4)、백개소10(interleukin 10,IL-10)화γ-간우색(interferon-γ,IFN -γ)적영향,대천명PGE2통과조공ASMCs분비Th1/Th2세포인자대기도염증발휘작용궤제유중요의의.방법 용대서기도평활기세포주여종농도위10 -9～10-6적PGE2공배양24 h,분6조,공4개농도제도.용Annexin V EGFP/PI쌍염세포조망검측시제합재류식세포의측정조망솔,동시용IL-4、IL- 10화IFN-γ ELISA시제합재매표의검측기농도.매4개복공위1조,실험중복2차,수거위8공적평균치.결과 응용불동농도적PGE2후ASMCs각조세포조망솔여대조조상비무명현변화.단IL분비여PGE2유현저적량효관계,IL-4화IL-10수PGE2농도하강이분비량현저하강,단IFN-γ수PGE2농도하강이분비량정승고추세,량자정분리현상(공백대조、음성대조、10-9 PGE2、10-8 PGE2、10 -7 PGE2、10 -6 PGE2조,IL- 4분별위6.77 ± 1.58、9.24±2.45、38.17±11.33、29.27±11.12、26.05±9.49、21.11±7.21;IL-10위4.8±1.06、6.35±2.11、71.89±12.03、43.68 ±11.25、28.89± 8.7여、24.31±6.67；IFN-γ위2.17±1.97、3.25±1.48、10.63±4.75、13.4±5.57、15.47±6.16、19.54±6.35).결론 PGE2능조공ASMCs분비세포인자,수착농도제도증가능촉진Th1형세포인자IFN-γ분비,동시억제Th2형세포인자 IL-4、IL-10분비,단대ASMCs조망무현선영향.인위재정상상태하 PGE2적분비여유지기도내Th1/Th2적평형유관.
Objective Airway remodeling is closely related to migration,proliferation,secretion function of airway smooth muscle cells ( ASMCs ). Prostaglandin E2 (PGE2) have a regulatory role in these functions.To culture with different concentration gradient of PGE2 and ASMCs in vitro and to study ASMCs secretion functions of interleukin 4 (IL -4) and interleukin 10 (IL -10) and Interferon-γ (IFN- γ),and apoptosis of ASMCs,this is favorable to understand ASMCs immune regulation mechanism in asthma,and the regulatiun role of PGE2.Methods The rat airway smooth muscle cells and the final concentration of 10 -9～ 10 -6 PGE2 were cultured for 24 hours,divided into 6 groups,and 4 concentration gradient. Flow cytometric were used determination of apoptosis with Annexin V-EGFP/PI double staining apoptosis detection kit,United States Bio-Rad 680 enzyme mark instrument were used detection of its concentration with IL- 10,IL- 4,IFN- γ ELISA kit.4 complex holes into 1 groups,the experiment was repeaied 2 times,average data for 8 holes.Results There were no significant changes in ASMCs cell apoptosis rate after application of the different concentrations of PGE2 compared with the control group.But interleukin secretion and PGE2 has significant dose-effect relationship.When PGE2 concentration were decreased,the secretion of IL- 4 and IL- 10 decreased significantly,but the IFN- γ secretion was increased.(Control,negative,10- 9 PGE2,10- 8 PGE2,10 -7 PGE2,10 -6 PCE2 group,IL- 4 was respectively 6.77 ±1.58,9.24 ± 2.45,38.17 ±11.33,29.27 ±11.12,26.05 ±9.49,21.11 ± 7.21;IL-10 was 4.8 ± 1.06,6.35±2.11,71.89 ±12.03,43.68 ±11.25,28.89 ±8.75,24.31 ± 6.67; 2.17 ± 1.97,IFN-γ 3.25± 1.48,10.63 ± 4.75,13.4 ± 5.57,15.47 ± 6.16,19.54 ± 6.35).Conclusions PGE2 can regulate the secretion of ASMCs cytokines,increased with concentration gradient can promote Th1 cytokines IFN-γ increased secretion,while inhibition of type Th2 cytokine secretion of IL-10,IL-4,but no significant effect on the apoptosis of ASMCs. We think the key role of the normal state of PGE2 secretion were related to maintaining Th1/Th2 balance in the airway.
