中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
Chinese Journal of Industrial Hygiene and Occupational Diseases
2015年
9期
652-657
,共6页
魏明%涂玲%梁颖红%刘佳%龚艳杰%张宜花%杨璐
魏明%塗玲%樑穎紅%劉佳%龔豔傑%張宜花%楊璐
위명%도령%량영홍%류가%공염걸%장의화%양로
丙烯醛%白细胞介素-17%T淋巴细胞
丙烯醛%白細胞介素-17%T淋巴細胞
병희철%백세포개소-17%T림파세포
Acrolein%Intcerleukin-17%T-lymphocytes
目的 观察丙烯醛染毒大鼠外周血与支气管肺泡灌洗液(BALF)中CD4+白细胞介素(IL)-17表达阳性T细胞(Th17细胞)与CD4+Foxp3表达阳性调节性T细胞(Treg细胞)及相关因子水平变化,探讨Th17细胞和Treg细胞在丙烯醛诱导气道炎症发生中的作用.方法 将40只健康清洁级雄性Wistar大鼠分为染毒2、4周组和对照2、4周组,每组10只.染毒2、4周组雾化吸入4×10-6 kg/L的丙烯醛,雾化时间为每天3次,每次2h,分别持续2和4周;对照2、4周组雾化吸入生理盐水,雾化时间为每天3次,每次2h,分别持续2和4周.收集BALF进行细胞学计数和分类,采用酶联免疫吸附法检测大鼠血清和BALF上清液中IL-17和IL-6水平,用流式细胞术检测Th17细胞和Treg细胞比例,用实时荧光定量PCR法检测IL-17mRNA和Foxp3 mRNA的表达.多组间比较采用单因素方差分析,组内两两比较采用SNK法和Games-Howell法.结果 染毒2和4周组大鼠外周血中IL-17浓度分别为[(52.64±1.89)和(76.73±5.57) ng/L],BALF中IL-17浓度分别为[(79.07±5.67)和(96.61±6.44) ng/L],均明显高于对照2周组[(40.05±3.12)和(56.75 ±4.37) ng/L]及4周组[(38.75±3.23)和(53.27±4.48) ng/L];染毒4周组与其他3组比较,差异均有统计学意义(均P＜0.01).染毒4周组大鼠外周血中IL-6浓度为(31.57±2.25)ng/L,明显高于对照4周组(10.98±0.49) ng/L;染毒2和4周组大鼠BALF中IL-6浓度分别为[(33.28±2.27)和(46.24±3.16) ng/L],明显高于对照2和4周组[(16.37±].49)和(17.02±1.43) ng/L].染毒2和4周组大鼠外周血中Th17细胞比例分别为[(1.82±0.18)％和(3.75±0.48)％],BALF中Th17细胞比例分别为[(7.23±0.27)％和(8.12±0.38)％],均明显高于对照2周组[(0.96±0.07)％和(5.64±0.63)％]及4周组[(1.01±0.08)％和(5.86±0.57)％];染毒4周组与其他3组比较,差异均有统计学意义(均P＜0.01).染毒2和4周组大鼠BALF中Treg细胞比例分别为[(8.83±0.52)％和(12.05±0.74)％],均明显高于对照2和4周组[(4.37±0.27)％和(5.01±0.37)％].染毒2和4周组大鼠外周血中IL-17 mRNA表达量分别为[(25.78±2.31)和(34.69±2.01)],BALF中IL-17 mRNA表达量分别为[(23.04±1.78)和(34.56±3.12)],均明显高于对照2周组[(11.04±2.53)和(11.08±2.05)]及4周组[(12.03±2.34)和(12.69±2.69)];染毒2和4周组大鼠BALF中Foxp3 mRNA表达量分别为[(26.37±3.24)和(33.19±2.98)],明显高于对照2和4周组[(12.37±2.56)和(13.12±3.08)].染毒组大鼠BALF中Th17细胞与其BALF中细胞总数和巨噬细胞数呈正相关(r值分别为0.5126和0.5437,均P＜0.01).结论 丙烯醛暴露可导致大鼠Th17细胞和Treg细胞及相关炎症因子水平发生变化,Treg细胞可能参与气道炎症的免疫调节,Th17细胞异常升高可能与大鼠气道炎症反应的发生及持续进展有关.
目的 觀察丙烯醛染毒大鼠外週血與支氣管肺泡灌洗液(BALF)中CD4+白細胞介素(IL)-17錶達暘性T細胞(Th17細胞)與CD4+Foxp3錶達暘性調節性T細胞(Treg細胞)及相關因子水平變化,探討Th17細胞和Treg細胞在丙烯醛誘導氣道炎癥髮生中的作用.方法 將40隻健康清潔級雄性Wistar大鼠分為染毒2、4週組和對照2、4週組,每組10隻.染毒2、4週組霧化吸入4×10-6 kg/L的丙烯醛,霧化時間為每天3次,每次2h,分彆持續2和4週;對照2、4週組霧化吸入生理鹽水,霧化時間為每天3次,每次2h,分彆持續2和4週.收集BALF進行細胞學計數和分類,採用酶聯免疫吸附法檢測大鼠血清和BALF上清液中IL-17和IL-6水平,用流式細胞術檢測Th17細胞和Treg細胞比例,用實時熒光定量PCR法檢測IL-17mRNA和Foxp3 mRNA的錶達.多組間比較採用單因素方差分析,組內兩兩比較採用SNK法和Games-Howell法.結果 染毒2和4週組大鼠外週血中IL-17濃度分彆為[(52.64±1.89)和(76.73±5.57) ng/L],BALF中IL-17濃度分彆為[(79.07±5.67)和(96.61±6.44) ng/L],均明顯高于對照2週組[(40.05±3.12)和(56.75 ±4.37) ng/L]及4週組[(38.75±3.23)和(53.27±4.48) ng/L];染毒4週組與其他3組比較,差異均有統計學意義(均P＜0.01).染毒4週組大鼠外週血中IL-6濃度為(31.57±2.25)ng/L,明顯高于對照4週組(10.98±0.49) ng/L;染毒2和4週組大鼠BALF中IL-6濃度分彆為[(33.28±2.27)和(46.24±3.16) ng/L],明顯高于對照2和4週組[(16.37±].49)和(17.02±1.43) ng/L].染毒2和4週組大鼠外週血中Th17細胞比例分彆為[(1.82±0.18)％和(3.75±0.48)％],BALF中Th17細胞比例分彆為[(7.23±0.27)％和(8.12±0.38)％],均明顯高于對照2週組[(0.96±0.07)％和(5.64±0.63)％]及4週組[(1.01±0.08)％和(5.86±0.57)％];染毒4週組與其他3組比較,差異均有統計學意義(均P＜0.01).染毒2和4週組大鼠BALF中Treg細胞比例分彆為[(8.83±0.52)％和(12.05±0.74)％],均明顯高于對照2和4週組[(4.37±0.27)％和(5.01±0.37)％].染毒2和4週組大鼠外週血中IL-17 mRNA錶達量分彆為[(25.78±2.31)和(34.69±2.01)],BALF中IL-17 mRNA錶達量分彆為[(23.04±1.78)和(34.56±3.12)],均明顯高于對照2週組[(11.04±2.53)和(11.08±2.05)]及4週組[(12.03±2.34)和(12.69±2.69)];染毒2和4週組大鼠BALF中Foxp3 mRNA錶達量分彆為[(26.37±3.24)和(33.19±2.98)],明顯高于對照2和4週組[(12.37±2.56)和(13.12±3.08)].染毒組大鼠BALF中Th17細胞與其BALF中細胞總數和巨噬細胞數呈正相關(r值分彆為0.5126和0.5437,均P＜0.01).結論 丙烯醛暴露可導緻大鼠Th17細胞和Treg細胞及相關炎癥因子水平髮生變化,Treg細胞可能參與氣道炎癥的免疫調節,Th17細胞異常升高可能與大鼠氣道炎癥反應的髮生及持續進展有關.
목적 관찰병희철염독대서외주혈여지기관폐포관세액(BALF)중CD4+백세포개소(IL)-17표체양성T세포(Th17세포)여CD4+Foxp3표체양성조절성T세포(Treg세포)급상관인자수평변화,탐토Th17세포화Treg세포재병희철유도기도염증발생중적작용.방법 장40지건강청길급웅성Wistar대서분위염독2、4주조화대조2、4주조,매조10지.염독2、4주조무화흡입4×10-6 kg/L적병희철,무화시간위매천3차,매차2h,분별지속2화4주;대조2、4주조무화흡입생리염수,무화시간위매천3차,매차2h,분별지속2화4주.수집BALF진행세포학계수화분류,채용매련면역흡부법검측대서혈청화BALF상청액중IL-17화IL-6수평,용류식세포술검측Th17세포화Treg세포비례,용실시형광정량PCR법검측IL-17mRNA화Foxp3 mRNA적표체.다조간비교채용단인소방차분석,조내량량비교채용SNK법화Games-Howell법.결과 염독2화4주조대서외주혈중IL-17농도분별위[(52.64±1.89)화(76.73±5.57) ng/L],BALF중IL-17농도분별위[(79.07±5.67)화(96.61±6.44) ng/L],균명현고우대조2주조[(40.05±3.12)화(56.75 ±4.37) ng/L]급4주조[(38.75±3.23)화(53.27±4.48) ng/L];염독4주조여기타3조비교,차이균유통계학의의(균P＜0.01).염독4주조대서외주혈중IL-6농도위(31.57±2.25)ng/L,명현고우대조4주조(10.98±0.49) ng/L;염독2화4주조대서BALF중IL-6농도분별위[(33.28±2.27)화(46.24±3.16) ng/L],명현고우대조2화4주조[(16.37±].49)화(17.02±1.43) ng/L].염독2화4주조대서외주혈중Th17세포비례분별위[(1.82±0.18)％화(3.75±0.48)％],BALF중Th17세포비례분별위[(7.23±0.27)％화(8.12±0.38)％],균명현고우대조2주조[(0.96±0.07)％화(5.64±0.63)％]급4주조[(1.01±0.08)％화(5.86±0.57)％];염독4주조여기타3조비교,차이균유통계학의의(균P＜0.01).염독2화4주조대서BALF중Treg세포비례분별위[(8.83±0.52)％화(12.05±0.74)％],균명현고우대조2화4주조[(4.37±0.27)％화(5.01±0.37)％].염독2화4주조대서외주혈중IL-17 mRNA표체량분별위[(25.78±2.31)화(34.69±2.01)],BALF중IL-17 mRNA표체량분별위[(23.04±1.78)화(34.56±3.12)],균명현고우대조2주조[(11.04±2.53)화(11.08±2.05)]급4주조[(12.03±2.34)화(12.69±2.69)];염독2화4주조대서BALF중Foxp3 mRNA표체량분별위[(26.37±3.24)화(33.19±2.98)],명현고우대조2화4주조[(12.37±2.56)화(13.12±3.08)].염독조대서BALF중Th17세포여기BALF중세포총수화거서세포수정정상관(r치분별위0.5126화0.5437,균P＜0.01).결론 병희철폭로가도치대서Th17세포화Treg세포급상관염증인자수평발생변화,Treg세포가능삼여기도염증적면역조절,Th17세포이상승고가능여대서기도염증반응적발생급지속진전유관.
Objective To evaluate the changes of CD4+ IL-17+T (Th17)and CD4+Foxp3+regulatory T (Treg) cells in peripheral blood and bronchoalveolar lavage fluid (BALF),and therefore to explore the role of Th 17 and Treg in acrolein exposure airway inflammation in rats.Methods Forty male Wistar rats were randomly divided into 4 groups:a 2 wk acrolein exposure group,a 4 wk acrolein exposure group,a 2 wk control group and a 4 wk control group (n=10 each).Cells in BALF were collected and analyzed by absolute and differential cell counts.IL-17 and IL-6 levels in serum and BALF were tested by enzyme linked immunosorbent assay(ELISA).The proportion of CD4+IL-17+T and CD4+ Foxp3+Treg in peripheral blood and BALF were determined by flow cytometry.The mRNA expressions of IL-17 and Foxp3 were measured by real-time PCR.Comparisons of the data between different groups were performed using one-way ANOVA,and SNK and Games-Howell test were used for comparison between 2 groups.Results Levels of IL-17 were remarkable increased in the 2 wk acrolein exposure group and the 4 wk acrolein exposure group in serum [(52.64±1.89) ng/L,(76.73±5.57) ng/L],and BALF [(79.07 ±5.67) ng/L,(96.61±6.44) ng/L] compared with the 2 wk control group [(40.05±3.12) ng/L,(56.75±4.37) ng/L] and the 4 wk control group [(38.75±3.23) ng/L,(53.27±4.48)ng/L],all P＜0.01.IL-6 was increased in the 2 wk and the 4 wk acrolein exposure group [(33.28±2.27) ng/L,(46.24±3.16) ng/L] compared with the 2 wk and the 4 wk control group [(16.37±1.49) ng/L,(17.02±1.43) ng/L] in BALF.Ratio of Th 17 was higher in the 2 wk and the 4 wk acrolein exposure groups in peripheral blood(1.82±0.18)％,(3.75±0.48)％and BALF [(7.23±0.27)％,(8.12±0.38)％] compared with the 2 wk [(0.96±0.07)％,(5.64±0.63)％] and the 4 wk control group [(1.01 ±0.08)％,(5.86±0.57)％].Ratio of Treg in BALF was higher in the acrolein exposure groups [(8.83±0.52)％,(12.05±0.74)％] compared with the control groups [(4.37±0.27)％,(5.01±0.37)％].The level of IL-17 mRNA was increased in the 2 wk and the 4 wk acrolein exposure group in peripheral blood [(25.78±2.31),(34.69±2.01)] and in BALF [(23.04±1.78),(34.56±3.12)] compared with the 2 wk [(11.04±2.53),(11.08±2.05)] and the 4 wk [(12.03±2.34),(12.69±2.69)] control groups.Foxp3 mRNA was increased in the acrolein exposure groups [(26.37±3.24),(33.19±2.98)] (24.4±2.7),(30.3±2.7) compared with the control groups [(12.37±2.56),(13.12±3.08)].Th 17 in acrolein exposure groups was positively correlated with counts of total cells and macrophages (r=0.5126,0.5437,all P＜0.01).Conclusions A changed expression of Th17 and Treg cells and an vary of inflammatory cytokines were evident in airway inflammation of acrolein exposed rats,suggesting that Treg was involved in the immunological regulation and Th17 was associated with the persistent inflammation in acrolein induced airway inflammation in rats.
