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
2001年
1期
30-33
,共4页
二氧化硅%间隙连接%肺纤维化%FRAP技术
二氧化硅%間隙連接%肺纖維化%FRAP技術
이양화규%간극련접%폐섬유화%FRAP기술
目的 探索矽肺发病过程中二氧化硅(SiO2)刺激肺纤维化发生与肺成纤维细胞间隙连接通讯功能 (GJIC)下调的关系。方法 不同剂量的SiO2刺激SD大鼠肺泡巨噬细胞(PAM)及具有肺泡巨噬细胞特性的、经12-O-四 葵酰基-佛波-13-乙酸酯(佛波酯,PMA或TPA)诱导分化的人血单核细胞株(pTHP-1)培养 上清液 ,作用于中国仓鼠肺成纤维(CHL)细胞。用四唑盐(MTT)法测定CHL细胞增殖(A570 n m);用激光扫描共聚焦显微镜,按荧光光漂白后再恢复(FRAP)技术测定CHL细胞GJIC功能[ 荧光传递速率K,(×10-3/s)]。结果 SiO2刺激的PAM上清液和pTHP-1上清液均能诱导CHL细胞增殖(PAM:F=3.205, P<0.05;pTHP-1:F=13.779,P<0.01)及抑制GJIC功能(PAM:F=19.948, P<0.01;pTHP-1:F=9.365,P<0.01),并随SiO2浓度增加(50、100、20 0和500 μg/ml),细胞增殖能力渐强,呈剂量-效应关系(PAM:r=-0.803,P<0 .05;pTHP-1:r=-0.914,P<0.01)。PAM和pTHP-1细胞上清液可以增强CHL细胞 的GJIC功能(K均数分别为21.24×10-3/s、18.92×10-3/s)和抑 制细胞增殖(A 570 nm均数分别为0.506、0.218),和空白对照(7.81×10-3/s、7 .81×10-3/s和0.539、0.388)相比,差异均有显著性(P<0.05)。结论 SiO2刺激PAM和pTHP-1培养上清液可以抑制肺成纤维细胞的GJIC功能,并诱导细胞增殖。 推测在矽肺纤维化发生过程中,肺成纤维细胞GJIC功能的下调进而诱导肺成纤维细胞增殖 可能发挥了重要的作用。
目的 探索矽肺髮病過程中二氧化硅(SiO2)刺激肺纖維化髮生與肺成纖維細胞間隙連接通訊功能 (GJIC)下調的關繫。方法 不同劑量的SiO2刺激SD大鼠肺泡巨噬細胞(PAM)及具有肺泡巨噬細胞特性的、經12-O-四 葵酰基-彿波-13-乙痠酯(彿波酯,PMA或TPA)誘導分化的人血單覈細胞株(pTHP-1)培養 上清液 ,作用于中國倉鼠肺成纖維(CHL)細胞。用四唑鹽(MTT)法測定CHL細胞增殖(A570 n m);用激光掃描共聚焦顯微鏡,按熒光光漂白後再恢複(FRAP)技術測定CHL細胞GJIC功能[ 熒光傳遞速率K,(×10-3/s)]。結果 SiO2刺激的PAM上清液和pTHP-1上清液均能誘導CHL細胞增殖(PAM:F=3.205, P<0.05;pTHP-1:F=13.779,P<0.01)及抑製GJIC功能(PAM:F=19.948, P<0.01;pTHP-1:F=9.365,P<0.01),併隨SiO2濃度增加(50、100、20 0和500 μg/ml),細胞增殖能力漸彊,呈劑量-效應關繫(PAM:r=-0.803,P<0 .05;pTHP-1:r=-0.914,P<0.01)。PAM和pTHP-1細胞上清液可以增彊CHL細胞 的GJIC功能(K均數分彆為21.24×10-3/s、18.92×10-3/s)和抑 製細胞增殖(A 570 nm均數分彆為0.506、0.218),和空白對照(7.81×10-3/s、7 .81×10-3/s和0.539、0.388)相比,差異均有顯著性(P<0.05)。結論 SiO2刺激PAM和pTHP-1培養上清液可以抑製肺成纖維細胞的GJIC功能,併誘導細胞增殖。 推測在矽肺纖維化髮生過程中,肺成纖維細胞GJIC功能的下調進而誘導肺成纖維細胞增殖 可能髮揮瞭重要的作用。
목적 탐색석폐발병과정중이양화규(SiO2)자격폐섬유화발생여폐성섬유세포간극련접통신공능 (GJIC)하조적관계。방법 불동제량적SiO2자격SD대서폐포거서세포(PAM)급구유폐포거서세포특성적、경12-O-사 규선기-불파-13-을산지(불파지,PMA혹TPA)유도분화적인혈단핵세포주(pTHP-1)배양 상청액 ,작용우중국창서폐성섬유(CHL)세포。용사서염(MTT)법측정CHL세포증식(A570 n m);용격광소묘공취초현미경,안형광광표백후재회복(FRAP)기술측정CHL세포GJIC공능[ 형광전체속솔K,(×10-3/s)]。결과 SiO2자격적PAM상청액화pTHP-1상청액균능유도CHL세포증식(PAM:F=3.205, P<0.05;pTHP-1:F=13.779,P<0.01)급억제GJIC공능(PAM:F=19.948, P<0.01;pTHP-1:F=9.365,P<0.01),병수SiO2농도증가(50、100、20 0화500 μg/ml),세포증식능력점강,정제량-효응관계(PAM:r=-0.803,P<0 .05;pTHP-1:r=-0.914,P<0.01)。PAM화pTHP-1세포상청액가이증강CHL세포 적GJIC공능(K균수분별위21.24×10-3/s、18.92×10-3/s)화억 제세포증식(A 570 nm균수분별위0.506、0.218),화공백대조(7.81×10-3/s、7 .81×10-3/s화0.539、0.388)상비,차이균유현저성(P<0.05)。결론 SiO2자격PAM화pTHP-1배양상청액가이억제폐성섬유세포적GJIC공능,병유도세포증식。 추측재석폐섬유화발생과정중,폐성섬유세포GJIC공능적하조진이유도폐성섬유세포증식 가능발휘료중요적작용。
Objective To explore the relationships betw een pulmonary fibrosis and downregulation of gap junctional intercellular comm unication(GJIC) of the fibroblasts after stimulation by SiO2. Metho ds The pulmonary alveolar macrophage(PAM) and the phorbol 12-myrist ate 13-acetate(PMA or TPA) primed THP-1(pTHP-1),a monocyte-like cell line wi th the properties of PAM,were incubated in the serum-free RPMI?1640 containing SiO2 at various concentrations.The proliferation of Chinese hamster lung(CHL) fibroblast induced by cultured PAM or pTHP supernatant was detected by using MT T assay(to show as the a bsorbency,A570 nm),and GJIC between those cells was measured by using the fluorescence redis tribution after photobleaching(FRAP) assay(to show as the transfer rate of the f luorescence,K×10-3/s) performed with a laser scanning confocal micr oscope(LSCM,Carl Zeiss LSM 510,release 2.01). Results Bot h SiO2 exposed PAM and pTHP-1 supern atants could induce the proliferation(PAM:F=3.205,P<0.05;pTHP-1:F=1 3.779,P<0.01) and the downregulation of GJIC(PAM:F=19.948,P<0. 01;pTHP-1:F=9.365,P<0.01) of the CHL cells.In the range of 0,50,1 00,200 and 500 μg/ml SiO2 concentrations,the proliferation(A570 nm values) and GJIC(the transfer rate,K)were fitted well in the dose-effect re lationship(PAM:r=-0.803,P<0.05;pTHP-1:r=-0.914,P<0.01). Compared with the blank control,both PAM and pTHP-1 supernatants could upregu late GJIC(K:21.24×10-3/s,18.92×10-3/s vs. 7.81×10 -3/s,7.81×10-3/s respectively,P<0.05) and inh ibit the proliferation of CHL cell(A570 nm:0.506,0.218 vs. 0.5 39,0.388 respectively,P<0.05). Conclusion T hrough the way of stimulating PAM,SiO2 could inhibit GJIC of fibroblasts,and induce fibroblast proliferation.In the pathoge nesis of silicotic fibrosis,the downregulation of GJIC of the pulmonary fibrobla sts may play an important role.