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
2009年
7期
395-399
,共5页
朱兰%梁永强%马小兵%王献华%孙树勋
硃蘭%樑永彊%馬小兵%王獻華%孫樹勛
주란%량영강%마소병%왕헌화%손수훈
二氧化硅%巨噬细胞%成纤维细胞%基质金属蛋白酶%胶原
二氧化硅%巨噬細胞%成纖維細胞%基質金屬蛋白酶%膠原
이양화규%거서세포%성섬유세포%기질금속단백매%효원
Silicon dioxide%Macrophages%Fibroblasts%Matrix metalloproteinases%Collagen
目的 探讨经SiO2刺激的肺泡巨噬细胞(AM)通过人胚肺成纤维细胞(HELF)对基质金属蛋白酶-1(MMP-1)及其抑制因子(TIMP-1)和Ⅲ型胶原表达的影响.方法 收集矽肺患者AM,将其分为加入SiO2粉尘悬液的处理组和仅加入无血清培养液的对照组.培养18 h后吸出培养上清.将原代培养的HELF分为4组:(1)处理组:加入经SiO2刺激的AM上清;(2)对照组:加人未经SiO2刺激的AM上清;(3)10%血清组:仅加入含体积分数为10%胎牛血清的DMEM;(4)空白对照组:仅加入含体积分数为3%胎牛血清的DMEM.4组均分别于6、12、18、24、36、48 h取出细胞爬片,吸出上清,用免疫细胞化学方法 检测HELF中MMP-1和TIMP-1的表达,用Western blot法检测HELF条件培养上清中Ⅲ型胶原的表达.结果 处理组18、24、36、48 h MMP-1表达分别为0.0605±0.0201,0.0519±0.0117,0.0412±0.0105,0.0213±0.0106,较对照组和空白对照组明显降低,差异均有统计学意义(P<0.05,P<0.01).与对照组和空白对照组相比,处理组各时间点TIMP-1表达和Ⅲ型胶原含量明显增高,差异均有统计学意义(P<0.05,P<0.01),TIMP-1/MMP-1与Ⅲ型胶原呈正相关(r=0.88,P<0.01).结论 SiO2经AM的介导可促进FB表达TIMP-1和Ⅲ型胶原,抑制HELF表达TIMP-1,TIMP-1/MMP-1的失衡与Ⅲ型胶原的病理性累积有关.
目的 探討經SiO2刺激的肺泡巨噬細胞(AM)通過人胚肺成纖維細胞(HELF)對基質金屬蛋白酶-1(MMP-1)及其抑製因子(TIMP-1)和Ⅲ型膠原錶達的影響.方法 收集矽肺患者AM,將其分為加入SiO2粉塵懸液的處理組和僅加入無血清培養液的對照組.培養18 h後吸齣培養上清.將原代培養的HELF分為4組:(1)處理組:加入經SiO2刺激的AM上清;(2)對照組:加人未經SiO2刺激的AM上清;(3)10%血清組:僅加入含體積分數為10%胎牛血清的DMEM;(4)空白對照組:僅加入含體積分數為3%胎牛血清的DMEM.4組均分彆于6、12、18、24、36、48 h取齣細胞爬片,吸齣上清,用免疫細胞化學方法 檢測HELF中MMP-1和TIMP-1的錶達,用Western blot法檢測HELF條件培養上清中Ⅲ型膠原的錶達.結果 處理組18、24、36、48 h MMP-1錶達分彆為0.0605±0.0201,0.0519±0.0117,0.0412±0.0105,0.0213±0.0106,較對照組和空白對照組明顯降低,差異均有統計學意義(P<0.05,P<0.01).與對照組和空白對照組相比,處理組各時間點TIMP-1錶達和Ⅲ型膠原含量明顯增高,差異均有統計學意義(P<0.05,P<0.01),TIMP-1/MMP-1與Ⅲ型膠原呈正相關(r=0.88,P<0.01).結論 SiO2經AM的介導可促進FB錶達TIMP-1和Ⅲ型膠原,抑製HELF錶達TIMP-1,TIMP-1/MMP-1的失衡與Ⅲ型膠原的病理性纍積有關.
목적 탐토경SiO2자격적폐포거서세포(AM)통과인배폐성섬유세포(HELF)대기질금속단백매-1(MMP-1)급기억제인자(TIMP-1)화Ⅲ형효원표체적영향.방법 수집석폐환자AM,장기분위가입SiO2분진현액적처리조화부가입무혈청배양액적대조조.배양18 h후흡출배양상청.장원대배양적HELF분위4조:(1)처리조:가입경SiO2자격적AM상청;(2)대조조:가인미경SiO2자격적AM상청;(3)10%혈청조:부가입함체적분수위10%태우혈청적DMEM;(4)공백대조조:부가입함체적분수위3%태우혈청적DMEM.4조균분별우6、12、18、24、36、48 h취출세포파편,흡출상청,용면역세포화학방법 검측HELF중MMP-1화TIMP-1적표체,용Western blot법검측HELF조건배양상청중Ⅲ형효원적표체.결과 처리조18、24、36、48 h MMP-1표체분별위0.0605±0.0201,0.0519±0.0117,0.0412±0.0105,0.0213±0.0106,교대조조화공백대조조명현강저,차이균유통계학의의(P<0.05,P<0.01).여대조조화공백대조조상비,처리조각시간점TIMP-1표체화Ⅲ형효원함량명현증고,차이균유통계학의의(P<0.05,P<0.01),TIMP-1/MMP-1여Ⅲ형효원정정상관(r=0.88,P<0.01).결론 SiO2경AM적개도가촉진FB표체TIMP-1화Ⅲ형효원,억제HELF표체TIMP-1,TIMP-1/MMP-1적실형여Ⅲ형효원적병이성루적유관.
Objective To study the effect of culture supernatant of alveolar macrophage alveolar macrophages(AM) stimulated by SiO2 on the expression of matrix metalloproteinases(MMP-1),tissue inhibitor of metalloproteinase-1 (TIMP-1) and collagen of fibroblast human embroyonie lung fibroblasts (HELF) in the de-velopment of silicosis fibrosis. Methods AMs were collected from a silicotic patient by bronehoalveolar lavage and exposed to SiO2, cultured human embryo lung fibroblast were allocated into a treated group, a con-trol group, a positive group, and a blank group. HELF was incubated with the cultured supernatant of AMs for 6, 12, 18, 24, 36, 48 h. Immunocytochemical and Western blot technology were used to detect MMP-1 and TIMP-1 expressions in HELF and collagen expression in supernatant of HELF respectively. Results The su-pernatant of AM exposed to SiO2 significantly decreased the expressions of MMP-1 (0.0605±0.0201, 0.0519±0.0117, 0.0412±0.0105 and 0.0213±0.0106 in the treated group at 18, 24, 36 and 48 h) compared with the control group and the blank group (P<0.05, P<0.01) but stimulated expressions of TIMP-1 and collagen (P< 0.05 ,P<0.01). The ratio of TIMP-1 to MMP-1 increased. The ratio of TIMP-1 to MMP-1 was positively corre-lated with the expression of collagen Ⅲ (r=0.88, P<0.01). Conclusion Through AM mediation SiO2 can ac-celerate the expression of TIMP-1 and collagen, and inhibit the expression of MMP-1. The imbalance between the expression of TIMP-1 and that of MMP-1 is related with the abnormal inerease in collagen Ⅲ.
