卡介苗对人肺腺上皮细胞表达Fall-39 mRNA及抗菌活性的影响

Effects of Fall-39 mRNA Expression and Antibacterial Activity of Human Pulmonary Gland Epithelial Cells Induced by BCG

目的 探讨卡介苗能否增强 Fall- 39在人肺腺上皮 SPC- A- 1细胞中的表达 ,分离鉴定卡介苗刺激作用的有效胞壁蛋白组份 ,并检测其抗菌活性。方法 根据 Gen Bank中人 Fall- 39的 c DNA序列资料 ,设计特异性引物 ,应用 RT- PCR法检测 SPC- A - 1细胞 Fall- 39m RNA的表达 ,采用超声破碎细胞、蔗糖密度梯度离心、SephadexG- 75柱层析等方法分离卡介苗胞壁蛋白质组份 ;采用琼脂糖弥散杀菌法检测 SPC- A- 1细胞培养上清的抗菌活性。结果 卡介苗刺激 SPC- A - 1细胞 Fall- 39m RNA的表达明显增强 ,而且这种诱导增强表达在一定范围内具有刺激物剂量和时间依赖性。卡介苗胞壁蛋白层析所得 6个组份中 ,组份 4 (相对分子质量范围约 2 1～ 2 9× 10 3)诱导 SPC-A- 1细胞 Fall- 39m RNA的表达增强约 8.5倍 ,其培养上清的抗菌活性显著增强。结论 BCG胞壁蛋白是 Fall- 39基因的激活因子 。

Objective To disclose whether BCG can enhance Fall 39 gene expression in human pulmonary gland epithelial cells, and to isolate and identify the bioactive proteins that stimulate Fall 39 gene expression in human pulmonary gland epithelial cells. Methods Fall 39 mRNA expression in SPC A 1 cells was detected by using RT PCR. The cell wall proteins were isolated by sucrose density gradient centrifugation and fractionated by Sephadex G 75 column chromatography. The antibacterial activity of the cell culture supernatant was determined by agarose radial diffusion assay. Results The enhanced expression of Fall 39 mRNA was dose and time dependent. BCG cell wall proteins fraction 4 had an activity that remarkably enhanced Fall 39 mRNA expression in SPC A 1 cells and augmented antibacterial activity of the cell culture supernatant. Conclusion These results indicated that BCG could stimulate Fall 39 mRNA expression in human pulmonary gland epithelial cells, thus providing an evidence base for shaping well a new strategy to enhance mucosa antibiotic peptide expression for the prevention and treatment of mucosal infections.