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
2014年
1期
24-29
,共6页
李雯曦%邹威凤%李冰%冉丕鑫
李雯晞%鄒威鳳%李冰%冉丕鑫
리문희%추위봉%리빙%염비흠
肺疾病,慢性阻塞性%木材%烟雾%上皮细胞
肺疾病,慢性阻塞性%木材%煙霧%上皮細胞
폐질병,만성조새성%목재%연무%상피세포
Pulmonary disease,chronic obstructive%Wood%Smog%Epithelial cells
目的 观察木材烟雾凝集物(WSC)对人支气管上皮细胞(HBEC)间充质转分化(EMT)样改变的影响.方法 WSC组和烟草烟雾凝集物(CSC)组分别用5、10、20、40和50 mg/L的WSC和CSC刺激HBEC 7 d,空白对照组仅用细胞培养基培养刺激HBEC 7 d,每组细胞数为1.2×106/ml.采用细胞计数试剂盒-8检测细胞总体活性.用WSC最大影响活性剂量(10 mg/L)刺激HBEC 7 d,观察细胞形态,用Western blot法和细胞免疫荧光技术检测EMT标记物E-钙黏蛋白、Ⅰ型胶原、波形蛋白和基质金属蛋白酶-9(MMP-9)的表达水平.多组间比较采用单因素方差分析,两组间比较采用t检验.结果 10 mg/L的WSC吸光度值最高(2.36 ±0.07),部分细胞形态由鹅卵石状向长梭形、多角形或星形改变,E-钙黏蛋白表达吸光度值(0.30 ±0.05)显著低于空白对照组(0.59±0.08),差异有统计学意义(F=22.07,P＜0.05),但与10 mg/L的CSC组吸光度值(0.30±0.05)比较,差异无统计学意义(F=22.07,P＞0.05)；10 mg/L的WSC组Ⅰ型胶原相对含量吸光度值(0.58±0.04)显著高于空白对照组(0.26±0.02),差异有统计学意义(F=119.72,P＜0.05),与CSC组(0.60±0.02)比较,差异无统计学意义(F=119.72,P＞0.05)；WSC组和CSC组MMP-9表达吸光度值(0.56±0.08和0.54±0.10)均显著高于空白对照组(0.19±0.03),差异均有统计学意义(F=21.79,均P＜0.05).结论 WSC能够刺激HBEC出现转分化样改变.
目的 觀察木材煙霧凝集物(WSC)對人支氣管上皮細胞(HBEC)間充質轉分化(EMT)樣改變的影響.方法 WSC組和煙草煙霧凝集物(CSC)組分彆用5、10、20、40和50 mg/L的WSC和CSC刺激HBEC 7 d,空白對照組僅用細胞培養基培養刺激HBEC 7 d,每組細胞數為1.2×106/ml.採用細胞計數試劑盒-8檢測細胞總體活性.用WSC最大影響活性劑量(10 mg/L)刺激HBEC 7 d,觀察細胞形態,用Western blot法和細胞免疫熒光技術檢測EMT標記物E-鈣黏蛋白、Ⅰ型膠原、波形蛋白和基質金屬蛋白酶-9(MMP-9)的錶達水平.多組間比較採用單因素方差分析,兩組間比較採用t檢驗.結果 10 mg/L的WSC吸光度值最高(2.36 ±0.07),部分細胞形態由鵝卵石狀嚮長梭形、多角形或星形改變,E-鈣黏蛋白錶達吸光度值(0.30 ±0.05)顯著低于空白對照組(0.59±0.08),差異有統計學意義(F=22.07,P＜0.05),但與10 mg/L的CSC組吸光度值(0.30±0.05)比較,差異無統計學意義(F=22.07,P＞0.05)；10 mg/L的WSC組Ⅰ型膠原相對含量吸光度值(0.58±0.04)顯著高于空白對照組(0.26±0.02),差異有統計學意義(F=119.72,P＜0.05),與CSC組(0.60±0.02)比較,差異無統計學意義(F=119.72,P＞0.05)；WSC組和CSC組MMP-9錶達吸光度值(0.56±0.08和0.54±0.10)均顯著高于空白對照組(0.19±0.03),差異均有統計學意義(F=21.79,均P＜0.05).結論 WSC能夠刺激HBEC齣現轉分化樣改變.
목적 관찰목재연무응집물(WSC)대인지기관상피세포(HBEC)간충질전분화(EMT)양개변적영향.방법 WSC조화연초연무응집물(CSC)조분별용5、10、20、40화50 mg/L적WSC화CSC자격HBEC 7 d,공백대조조부용세포배양기배양자격HBEC 7 d,매조세포수위1.2×106/ml.채용세포계수시제합-8검측세포총체활성.용WSC최대영향활성제량(10 mg/L)자격HBEC 7 d,관찰세포형태,용Western blot법화세포면역형광기술검측EMT표기물E-개점단백、Ⅰ형효원、파형단백화기질금속단백매-9(MMP-9)적표체수평.다조간비교채용단인소방차분석,량조간비교채용t검험.결과 10 mg/L적WSC흡광도치최고(2.36 ±0.07),부분세포형태유아란석상향장사형、다각형혹성형개변,E-개점단백표체흡광도치(0.30 ±0.05)현저저우공백대조조(0.59±0.08),차이유통계학의의(F=22.07,P＜0.05),단여10 mg/L적CSC조흡광도치(0.30±0.05)비교,차이무통계학의의(F=22.07,P＞0.05)；10 mg/L적WSC조Ⅰ형효원상대함량흡광도치(0.58±0.04)현저고우공백대조조(0.26±0.02),차이유통계학의의(F=119.72,P＜0.05),여CSC조(0.60±0.02)비교,차이무통계학의의(F=119.72,P＞0.05)；WSC조화CSC조MMP-9표체흡광도치(0.56±0.08화0.54±0.10)균현저고우공백대조조(0.19±0.03),차이균유통계학의의(F=21.79,균P＜0.05).결론 WSC능구자격HBEC출현전분화양개변.
Objective To observe the detrimental effects of wood smoke condensate (WSC) exposure on human bronchial epithelial cells (HBEC),and to explore the expression of epithelialmesenchymal transition (EMT) markers in HBEC exposed to WSC.Methods HBEC were exposed respectively to 5,10,20,40 and 50 mg/L of WSC/CSC for 7 days,with control groups only in cell culture medium at the same time,then the total cytoactivity was detected by cell counting kit-8.After observing the cellular morphology of WSC-stimulated HBEC.Western blot and immunofluorescence method were used to evaluate the expression levels of type Ⅰ collagen,vimentin,E-cad and MMP-9 in HBEC exposed to WSC (10 mg/L) and cigarette smoke condensate (CSC) (10 mg/L) for 7 days.Statistical evaluation of the continuous data was performed by ANOVA.Independent-Samples t-test for between-group comparisons.Results After 7 days of exposure to WSC,HBEC manifested a morphological characteristic of loss of cellcell contact and elongated shape.The level of E-cad was decreased in WSC exposure groups (Western blot:0.30 ± 0.05,F =22.07,P ＜ 0.05) compared with the groups without WSC exposure (Western blot:0.59 ± 0.08,F =22.07,P ＜ 0.05).In contrast,an upregulation in expression of type Ⅰ collagen (Western blot:0.58 ± 0.04 vs 0.26 ± 0.02,F =119.72,P ＜ 0.05) and MMP-9 (0.56 ± 0.08 vs 0.19 ± 0.03,F =21.79,P ＜ 0.05) was observed in the presence of WSC,compared with the control groups.Immunofluorescence analysis showed that after a 7-day exposure to WSC in these cells,the E-cad protein was lost whereas type Ⅰ collagen,vimentin and MMP-9 were acquired.Both Western blot and immunofluorescence analysis showed no difference in expression levels of E-cad,type Ⅰ collagen,vimentin and MMP-9 between WSC and CSC exposure groups.Conclusion WSC exposure could induce EMT-like process in human airway epithelial cells.
