高效解硅菌株代谢产物对二氧化硅暴露下CHL细胞的保护作用

Protective Effects of Silicate Bacterial Metabolites to CHL Cells Under Silica Exposures

目的观察高效解硅菌株代谢产物对中国仓鼠肺细胞在二氧化硅暴露下的保护作用。方法采集土壤、沙石标本,分离纯化其中的硅酸盐细菌;用液体培养基对高效解硅菌株进行培养,收集培养细菌12 d后的原液并过滤,滤液作为硅酸盐细菌代谢产物。取对数生长期的中国仓鼠肺细胞（CHL）,用胰酶消化后制成细胞悬液加入2个96孔培养板中培养。分为对照组、二氧化硅暴露组、硅酸盐细菌代谢产物组（根据不同菌株的产物,分为I、II 2个亚组）;二氧化硅＋硅酸盐细菌代谢产物组（根据不同菌株的产物,分为I、II 2个亚组）。对照组正常培养;二氧化硅暴露组仅在正常培养时加入二氧化硅粉尘;硅酸盐细菌代谢产物组正常培养时加入硅酸盐细菌代谢产物;二氧化硅＋硅酸盐细菌代谢产物组,在正常培养时加入二氧化硅粉尘后再加入硅酸盐细菌代谢产物;于给药24 h后采用4一甲偶氮唑蓝（MTT）法检测细胞存活情况。结果与二氧化硅暴露组比较,硅酸盐细菌代谢产物提取液I与SiO2体积比在3/7~9/1（硅酸盐细菌代谢产物提取液I剂量为30~90μl）时,吸光度（OD）值显著增加,差异有统计学意义（P〈0.01）;硅酸盐细菌代谢产物提取液II与SiO2体积比在2/8~9/1（硅酸盐细菌代谢产物提取液II剂量为20~90μl）时,OD值显著增加（P〈0.01）,且在20~50范围内,随菌液II体积增大,OD值递增,50~90范围内,随菌液II体积增大,OD值递减。结论硅酸盐细菌代谢产物对暴露于二氧化硅的CHL细胞有保护作用,并呈剂量依赖关系,且存在最佳保护作用的剂量范围。

Objective To observe the protective effects of silicate bacterial metabolites in Chinese hamster sources of lung（ CHL） cells in silica.Methods With the MTT assay,Chinese hamster lung cells（ CHL） cells in logarithmic growth were taken and were digested with trypsin;afterwards,the cell suspension was into to two 96-well culture plates for culturing.Cells were divided into control group,silica exposure group,silicate bacterial metabolic product group（ according to different strains of the product is divided into I,II,two subgroups） and silica ＆ silicate bacteria metabolite group（ divided into I,II sub-groups based on different products of the strains）.The control group were cultured in a normal way;the silica exposure group were added with the silica dust under normal culturing;silicate bacterial metabolites were added into the silicate bacteria metabolites group under normal culturing;the silica ＆ silicate bacteria metabolite group were added with silica dust in the normal culture before adding with the silicate bacterial metabolites.All the groups were administered 4 methyl thiazolyl tetrazolium（ MTT） assay to check the survival of cells 24h after dosage.Results Compared with the exposed group,when under condition of the volume ratio of silicate bacterial metabolite extract I and SiO2ranging from 3/7 to 9/1（ silicate bacterial metabolites extract dose at 30- 90μl）,the OD values increased significantly（ P ＆lt;0.01）;while under condition of the volume ratio of silicate bacterial metabolite extract II and SiO2ranging from 2/8 to 9/1（ silicate bacterial metabolites extract dose of 20-90μl）,the OD values increased significantly（ P＆lt;0.01）.Moreover,given the dose ranging 20-50μl,with the increasing volume of broth II,the OD values increased;while in 50-90μl range,the OD values decreased along with the increasing of broth II.Conclusion Silicate bacterial metabolites has a protective effect on CHL cells under the exposure of silicate and shows a dose-dependent relationship;meanwhile,a dose range exists related to the best protective effect.