游离二氧化硅诱导肺泡Ⅱ型上皮细胞转化机制

发布时间：2018-05-06 22:39

本文选题：游离二氧化硅 + 肺泡上皮细胞　；参考：《中国职业医学》2017年02期

【摘要】：目的观察游离二氧化硅(SiO_2)直接或间接作用于大鼠肺泡Ⅱ型上皮细胞株RLE-6TN而诱导发生上皮间质转化(EMT)的情况,及其对RLE-6TN表面标志蛋白表达影响情况。方法 (1)采用支气管肺泡灌洗术提取无特定病原体级SD大鼠肺泡巨噬细胞(AM)。以质量浓度为0~140 mg/L的SiO_2混悬液处理AM和RLE-6TN并培养24、48和72 h后,采用CCK-8实验检测细胞存活率,筛选后续实验染尘剂量。(2)取AM接种于Transwell的小室层与RLE-6TN共培养。设直接对照组(单独RLE-6TN,不作染尘处理)、直接染尘组(单独RLE-6TN,予质量浓度为100 mg/L的SiO_2混悬液处理)、间接对照组(AM与RLE-6TN共培养,不作染尘处理)和间接染尘组(AM与RLE-6TN共培养,予质量浓度为100 mg/L的SiO_2混悬液处理AM),分别于染尘0、24、48和72 h后收获细胞,用免疫印迹法测定RLE-6TN中的E-黏钙蛋白(E-cad)和α-平滑肌肌动蛋白(α-SMA)的相对表达水平。结果 (1)根据CCK-8结果,后续实验的SiO_2染尘剂量设为100 mg/L。(2)RLE-6TN中E-cad和α-SMA相对表达水平在染尘处理和染尘时间的交互效应上均有统计学意义(P0.01)。其中,直接染尘组和间接染尘组RLE-6TN在48和72 h的E-cad相对表达水平均低于同时间点直接对照组(P0.05),直接染尘组RLE-6TN在72 h时间点的E-cad相对表达水平分别低于同组0和24 h时间点(P0.05),间接染尘组RLE-6TN在48和72 h时间点的E-cad相对表达水平均低于同组0 h时间点(P0.05);在48和72 h时间点,直接染尘组和间接染尘组α-SMA相对表达水平分别高于同时间点的相应对照组(P0.05),且间接染尘组RLE-6TN中α-SMA相对表达水平均高于直接染尘组(P0.05);2个染尘组RLE-6TN中α-SMA相对表达水平均呈随着染尘时间的增加而增加的时间-效应关系(P0.05)。结论游离SiO_2直接或间接染尘均可诱导RLE-6TN发生EMT,导致E-cad表达下调,α-SMA表达呈现时间-效应性上调;间接染尘可能更易于诱导RLE-6TN发生EMT。
[Abstract]:Objective to observe the effect of free SiO-2) on the induction of epithelial mesenchymal transformation (EMTT) in rat alveolar epithelial cell line RLE-6TN directly or indirectly and its effect on the expression of RLE-6TN surface marker protein. Methods 1) the alveolar macrophages of SD rats were isolated by bronchoalveolar lavage. AM and RLE-6TN were treated with SiO_2 suspension of 0 ~ 140 mg/L and cultured for 24 h and 72 h. The survival rate of cells was detected by CCK-8 assay. The cell viability was detected by CCK-8 assay. The cells were inoculated in the compartment layer of Transwell and co-cultured with RLE-6TN. Direct control group (RLE-6TNN alone, without dust treatment, direct dust-exposed group (RLE-6TNN alone, treated with 100 mg/L SiO_2 suspension), indirect control group (RLE-6TNN) co-cultured with RLE-6TN and without Dust treatment) and indirect dust-exposed group (RLE-6TNN) were co-cultured with RLE-6TN. The cells were harvested after treatment with 100 mg/L SiO_2 suspension for 48 h and 72 h, respectively. The relative expression levels of E-cadand and 伪 -SMAin in RLE-6TN were determined by Western blot. Results (1) according to the CCK-8 results, the relative expression of E-cad and 伪 -SMA in the following experiment was set as 100 mg/L.(2)RLE-6TN. There was significant difference in the interaction between the time of dust exposure and the treatment time of E-cad and 伪 -SMA (P 0.01). Of which, The relative expression level of RLE-6TN in direct exposure group and indirect exposure group at 48 h and 72 h was lower than that in direct control group at the same time point (P0.05). The relative expression level of RLE-6TN in direct dust-exposed group at 72 h was lower than that in the same group at 0 h and 24 h, respectively. The relative expression level of RLE-6TN in the indirect dust-exposed group at 48 and 72 h was lower than that in the same group at 0 h and 72 h, and at 48 h and 72 h, the relative expression level of E-cad was lower than that in the same group at 0 h and 72 h, respectively. The relative expression level of 伪 -SMA in direct dust group and indirect dust group was higher than that in control group at the same time point, and the relative expression level of 伪 -SMA in RLE-6TN of direct dust group was higher than that of direct dust group, and the relative expression level of 伪 -SMA in RLE-6TN of two dust-exposed groups was higher than that of direct dust group. The relationship between time and effect increased with the increase of dust exposure time (P 0.05). Conclusion both direct and indirect exposure to free SiO_2 can induce the occurrence of EMTs in RLE-6TN, leading to down-regulation of E-cad expression and a time-dependent up-regulation of 伪 -SMA expression, and indirect dust exposure may be more likely to induce EMT-induced RLE-6TN.
【作者单位】：郑州大学公共卫生学院;郑州大学第三附属医院超声科;
【基金】：国家自然科学基金(81472954)
【分类号】：R135.2

【相似文献】