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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 effect of metabolites of high efficient lysozyme on Chinese hamster lung cells exposed to silica. Methods The samples of soil and gravel were collected and the silicate bacteria were isolated and purified. The highly efficient silicon-hydrolyzate strains were cultured in liquid medium. After 12 days of culture, the stock solution was collected and filtered, and the filtrate was used as the metabolite of silicate bacteria. Logarithmic growth phase of Chinese hamster lung cells (CHL), after digestion with trypsin made of cell suspension was added to two 96-well culture plates. Divided into control group, silica exposure group, silicate bacterial metabolite group (according to the product of different strains, divided into I, II 2 subgroups); silica + silicate bacterial metabolites group (according to different Strain products, divided into I, II 2 subgroups). The control group was cultured normally. Silica exposure group was added with silica dust only during normal culture. Silicate bacterial metabolites group was added with silicate bacterial metabolites during normal culture. Silica + silicate bacterial metabolites group , Adding silica dust in the normal culture and then adding the silicate bacterial metabolites; and measuring the cell viability by MTT assay 24 h after the administration. Results Compared with the silica exposure group, when the volume ratio of silicate bacteria metabolite extract I to SiO2 was in the range of 3/7 to 9/1 (the I dose of silicate bacterial metabolite extract was 30 ~ 90μl), the absorbance ( OD value increased significantly (P <0.01). The volume ratio of silicate bacterial extract II to SiO2 was in the range of 2 / 8-9 / 1 (the dose of silicate bacterial extract II 20 ~ 90μl), the OD value increased significantly (P <0.01), and in the range of 20 ~ 50, the OD value increased with the volume of bacterial liquid II increasing. Within the range of 50-90, OD value decreases. Conclusion Silicate bacterial metabolites have a protective effect on CHL cells exposed to silica in a dose-dependent manner with the optimal range of protection.