温石棉对大鼠肺泡巨噬细胞膜脂质过氧化的研究

温石棉对大鼠肺泡巨噬细胞膜脂质过氧化的研究肖国兵，马藻骅，邱华士宁波市卫生防疫站３１５０１０众所周知，长期接触石棉尘可引起石棉肺、肺癌等，但其致病机制尚不明了。已有试验表明石棉纤维可增加巨噬细胞脂质样化［１］。肺泡巨噬细胞作为肺纤维化的主要反应细胞，...     

二氧化硅和青石棉对小鼠肺泡巨噬细胞的影响

肺泡巨噬细胞(PAM),在炎症及免疫反应中起着重要作用,是肺的第一道防御屏障。目前,对PAM的吞噬功能方面研究较多,而对PAM体外集落形成能力方面报道较少。本文通过对小鼠经气管注入致纤维化粉尘(SiO_2和青石棉)及非致纤维化粉尘(TiO_2)后,着重观察其PAM体外集落形成能力的变化,以阐明吸入物质对PAM的增殖与分化的影响。 材料和方法 1.动物 C_3H/HeN纯系、健康雌性小鼠,6～7周龄。由日本SLC实验动物场提供。 2.实验粉尘 SiO_2(Min-u-sil;美国宾夕     

SiO2对人肺泡巨噬细胞核因子-κB表达的意义

目的：探讨二氧化硅（SiO2）对人肺泡巨噬细胞核因子-κB（nuclear factor-κB,NF-κB）表达的影响。方法：收集硅肺患者肺泡灌洗液中的肺泡巨噬细胞（AM），在体外再次染尘和无血清培养基：培养3、6、12、18、24、36h，用免疫组化检测AM中NF-κB的表达。结果：处理组（AM二次染尘，S组）NF-κB的表达明显上调，并且随刺激时间不同表达量不同，于18h达高峰。处理组NF-κB的表达在3、6、12、18、24、36h均高于对照组（ 矽肺患者AM，C组）。结论：SiO2可诱导AM的NF-κB的高表达，提示其在肺纤维化过程中起重要作用。     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

目的 探讨SiO2对矽肺患者的肺泡巨噬细胞(AM)基质金属蛋白酶-9(MMP-9)及金属蛋白酶组织抑制因子(TIMP)-1表达的影响.方法 收集矽肺患者肺泡AM,体外经SiO2刺激3、6、12、18、24、36 h,分别用明胶酶谱法和免疫细胞化学方法检测AM中TIMP-1、MMP-9蛋白表达和MMP-9活力.结果 实验组AM中6、12、18、24 h MMP-9的蛋白表达增强,在18 h时最高(积分吸光度值为0.386±0.037),与同期对照组相比,差异均有统计学意义(P＜0.05).实验组12、18、24、36h AM中MMP-9活力的表达增强,在24 h最强(吸光度值3.061±0.153),与同期对照组相比,差异均有统计学意义(P＜0.05).实验组与对照组各时间点TIMP-1蛋白表达比较,差异无统计学意义(P＞0.05).结论 体外SiO2刺激矽肺肺泡AM可影响MMP-9蛋白及其活力的表达.     

维生素C对肺泡巨噬细胞脂质过氧化及抗氧化酶的影响

目的 探讨维生素C拮抗镍所致细胞毒作用及作用机理。方法 采用体外细胞培养方法，观察维生素C对染镍肺泡巨噬细胞包膜脂质过氧化产物丙二醛（MDA）及超氧化物歧化酶（SOD）活性的影响。结果 在体外染镍肺泡巨噬细胞培养过程中加入不同浓度维生素C（25，50和100μmol/L）可提高肺泡噬细胞间SOD活性，具剂量反应关系。维生素C能显著抑制MDA生成量，其中以50和100μmol/L浓度组作用最为明显。结论 镍可致细胞脂质过氧化，维生素C具有拮抗镍所致细胞毒性的作用，可能与其抗氧化功能有关。     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

SiO2刺激矽肺肺泡巨噬细胞对MMP-9和TIMP-1表达的影响

Objective To investigate the effect of SiO2 on the expression of matrix metalloproteinase-9 (MMP-9)and tissue inhibitor of metalloproteinase-1 (TIMP-1)in human silicofic alveolar macrophages (AM).Methods Human alveolar macrophages were collected from a silicotic patient by bronchoalveolar lavage and exposed to silicon dioxide for 3h,6h,12h,18h,24h and 36h.The expression of the MMP-9 in the AM were detected with zymography and immunological method and the expression of the TIMP-1 in the AM with immunological method.Results The expressions of MMP-9 in the AM increased clearly along with the time,reached peak at 24 h when detected with zymography (average optical density:3.061±0.153 vs 2.851±0.164,P＜0.05);and reached peak at 18h when detected with immunological method (average optical density:0.386±0.037 vs 0.322±0.034,P＜0.05).The expression of the TIMP-1 in the AM did not vary when detected with immunological method (P＞0.05).Conclusion SiO2 may affect the expression of MMP-9 and MMP-9 activity in the cultured AM.     

硒对染尘肺泡巨噬细胞脂质过氧化和抗氧化酶影响的实验研究

目的探讨硒(Se)抗石英尘的细胞毒作用及其机理.方法采用体外细胞培养法,观察在不同浓度硒作用下,染石英尘的肺泡巨噬细胞(AM)膜脂质过氧化物丙二醛(MDA)生成量和AM内抗氧化酶-超氧化物歧化酶(SOD)活性.结果在体外染石英尘中加入硒有效抑制AM膜丙二醛生成量,其中以1.0×10-4mo/L硒作用最为明显,并可提高AM内超氧化物歧化酶活性,且有一定的剂量反应关系.结论硒具有抗石英尘对肺泡巨噬细胞脂质过氧化的作用.     

二氧化硅对矽肺患者肺泡巨噬细胞表皮生长因子表达的影响

目的探讨二氧化硅(SiO2)对矽肺患者肺泡巨噬细胞(AM)中表皮生长因子(EGF)表达的影响,以研究EGF对肺纤维化发生发展的影响。方法收集矽肺患者AM,加入含或不含SiO2的培养基离体培养3、6、12、18、24和36h,用免疫细胞化学法和酶联免疫吸附分析(ELISA)法检测AM细胞内和上清中EGF的表达。结果经SiO2粉尘刺激AM(处理组:S组)的EGF的表达较未经SiO2粉尘刺激AM(对照组:C组)的EGF表达量明显升高,差异具有统计学意义(P<0.05)。结论SiO可以促进矽肺患者AM表达EGF。     

二氧化硅对肺泡巨噬细胞膜K^＋通透性和LDH影响的研究

采用体外培养方法，检测了不同剂量的ＳｉＯ２和同一剂量，不同培养时间对肺泡巨噬细胞（ＡＭ）膜Ｋ含量和ＬＤＨ活性的影响。结果表明，低剂量的ＳｉＯ２，ＬＤＨ即有较明显的变化，且Ｋ含量和ＬＤＨ活性的剂量－反应关系和时间－效应关系。Ｋ含量和ＬＤＨ活性为反映膜损伤较简便和敏感的指标，为及时保护扫尘工人的健康提供实验依据。     

锌对SiO2致大鼠肺泡巨噬细胞氧化损伤的保护作用

目的 观察锌对SiO2致肺泡巨噬细胞（alveolar macrophages,AMs）产生氧自由基的动态变化和超微结构、膜酶的影响。方法 对AMs产生的O2^-和H2O2进行原位显示、定量测定并做相关趋势分析;以ATPase作为膜功能酶改变的反映;在亚细胞水平观察细胞形态结构。结果 与单纯SiO2组比较,锌保护组AMs O2^-和H2O2反应强度降低,超微结构及膜酶损伤减弱。结论 锌能减弱SiO2致AMs氧自由基的反应强度,降低超微结构和膜酶的损伤,保护AMs。其作用机制可能与锌增强AMs抗氧化作用有关,给临床用药提供了理论指导。