结核杆菌耐热抗原对A549细胞分泌SP-B和凋亡的影响

发布时间：2018-06-07 02:51

本文选题：结核杆菌耐热抗原 + A细胞　；参考：《安徽医科大学学报》2017年09期

【摘要】：目的探讨结核杆菌耐热抗原(Mtb-HAg)刺激诱导人肺腺癌细胞系(A549)细胞后对其肺泡表面活性物质相关蛋白B(SP-B)的分泌和凋亡的影响。方法用不同浓度的Mtb-HAg分别刺激诱导A549细胞,相同条件分别培养24、48 h,同时设置空白对照组(对照组1和对照组2)和阳性对照组[脂多糖(LPS)组和姜黄素组]。运用ELISA法检测对照组1、LPS组和实验组1(2、3μg/ml Mtb-HAg分别诱导A549细胞培养24 h和48 h)培养上清液中的SP-B表达量;使用实时荧光定量PCR法检测对照组1、LPS组和实验组1中基因SFTPB的相对表达量;采用流式细胞技术检测对照组2、姜黄素组和实验组2(2、3μg/ml Mtb-HAg分别诱导A549细胞培养24 h)中A549细胞的凋亡率。结果刺激诱导相同时间,实验组1的SP-B表达量低于对照组1,差异有统计学意义(P0.05);相同浓度刺激诱导随时间的延长,实验组1中SP-B表达量降低的不显著。相同培养时间,随着Mtb-HAg浓度的增加实验组1表达SP-B的基因SFTPB的相对表达量显著低于对照组1,差异有统计学意义(P0.05);而在相同有效刺激浓度下,其相对表达量随作用时间变化不显著。姜黄素组和实验组2中的A549细胞凋亡率显著高于对照组2(P0.05)。结论 Mtb-HAg对A549细胞表达SP-B的抑制作用明显,并能诱导A549细胞发生凋亡。
[Abstract]:Objective to investigate the effects of Mtb-HAg on the secretion and apoptosis of human lung adenocarcinoma cell line (A549) induced by Mtb-HAg. Methods A549 cells were stimulated with different concentrations of Mtb-HAg and cultured for 24 h under the same conditions. The control group (control group 1 and control group 2) and the positive control group [lipopolysaccharide (LPS) group and curcumin group] were set up at the same time. ELISA assay was used to detect the expression of SP-B in the supernatant of A549 cells cultured for 24 h and 48 h, respectively, and the relative expression of gene SFTPB in control group 1 and experimental group 1 was detected by real-time fluorescence quantitative PCR. Flow cytometry was used to detect the apoptosis rate of A549 cells induced by 2 渭 g/ml Mtb-HAg of curcumin group and 2 渭 g/ml Mtb-HAg of curcumin group for 24 h. Results the expression of SP-B in experimental group 1 was lower than that in control group 1 at the same time, the difference was statistically significant (P 0.05), but the expression of SP-B in experimental group 1 was not significantly decreased with the prolongation of stimulation at the same concentration. At the same culture time, the relative expression of SP-B gene SFTPB in experimental group 1 was significantly lower than that in control group 1 with the increase of Mtb-HAg concentration, and the difference was statistically significant (P 0.05). The relative expression level did not change significantly with the time of action. The apoptosis rate of A549 cells in curcumin group and experimental group 2 was significantly higher than that in control group 2 (P 0.05). Conclusion Mtb-HAg can inhibit the expression of SP-B in A549 cells and induce the apoptosis of A549 cells.
【作者单位】：安徽医科大学基础医学院免疫学教研室及过敏与免疫研究中心;
【基金】：国家自然科学基金面上项目(编号:81273245)
【分类号】：R378.911

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