摘要
目的研究罗勒多糖抑制转化生长因子-β(TGF-β)诱导的A549细胞上皮间质转化的作用,揭示罗勒多糖可能治疗特发性肺纤维化的机制。方法 A549细胞分为正常对照组、模型组(TGF-β5 ng·m L^(-1))、罗勒多糖组(TGF-β5 ng·m L^(-1)+罗勒多糖200μg·m L^(-1)),观察各组细胞形态学变化;实时荧光定量PCR(RT-q PCR)法检测各组Ⅰ型胶原蛋白(colⅠ)和上皮间质转化标志物表达;ELISA法检测各组细胞羟脯氨酸(HYP)浓度。结果与正常对照组比较,模型组细胞成纤维样变;与模型组比较,罗勒多糖组细胞维持上皮细胞形态。与正常对照组比较,模型组colⅠ基因表达明显上调(P<0.01),E-cadherin基因表达下调(P<0.01),Vimentin、α-SMA基因表达上调(P<0.05);与模型组相比,罗勒多糖组E-cadherin基因表达上调(P<0.05),Vimentin、α-SMA、colⅠ基因表达下调(P<0.05)。与正常对照组比较,模型组HYP含量增加(P<0.05);与模型组比较,罗勒多糖组HYP含量减少(P<0.05)。结论罗勒多糖能抑制TGF-β诱导的A549细胞上皮间质转化,其作用机制与下调colⅠ、Vimentin、α-SMA基因表达,上调E-cadherin基因表达,降低HYP含量有关,罗勒多糖可用于治疗特发性肺纤维化。
Objective To study the inhibitory effect of basil polysaccharide on epithelial-mesenchymal transition （EMT） induced by TGF- 13 in A549 cells, and to reveal the possible therapeutic mechanism of basil polysaccharide for idiopathic pulmonary fibrosis. Methods Cultured A549 cells were divided into three groups, normal control group, model group （TGF-β, 5 ng·mL-1） and basil polysaccharide group（TGF-β 5 ng ·mL-1 ＋ basil polysaccharide 200 μg· mL-1）. Morphological changes in A549 ceils were analyzed. Real-time quantitative polymerase chain reaction （RT-qPCR） was used to detect the expression of type I collagen （col I ） and EMT markers, and enzyme-linked immunosorbent assay （ELISA） was used for the examination of hydroxyproline content in A549 cells. Results Compared with the normal control group, the fibrosis was present in A549 cells of the model group, type I collagen gene expression level was obviously up-regulated （P 〈 0.01）, and the expression level of epithelial cell marker E-cadherin was down-regulated （P〈 0.01） while the levels of fibroblast cell marker Vimentin and myofibroblast marker ct-SMA were up-regulated （P 〈 0.05）. Compared with the model group, basil polysaccharide group had an effect on maintaining the morphology of the A549 cells, up-regulated E-cadherin gene expression （P〈 0.05）, down-regulated gene expression levels of Vimentin, ct-SMA and type I collagen（P〈0.05）. The content of hydroxyproline（P〈0.05） in the model group was higher than that of the normal control group, while the content of hydroxyproline in basil polysaecharide group cells was significantly lower （P 〈 0.05） than that in the model group. Conclusion Basil polysaccharide inhibited TGF-β-induced mesenchymal transition in the A549 cells, and the mechanism is related with the down-regulation of type I collagen, Vimentin, and α-SMA gene expression, with the up-regulation of E-cadherin expression, and also with the reduction of the content of hydroxyproline. It is indicated that basil polysaccharide may be a candidate drug for the treatment of idiopathic pulmonary fibrosis.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2017年第2期154-159,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家科技重大专项"重大新药创制"子课题(2014ZX09509001-001)
山东省高等学校科技计划项目(J14lK 09)
山东省优秀中青年科学家科研奖励基金(BS2011YY055)
关键词
罗勒多糖
特发性肺纤维化
上皮间质转化
Ⅰ型胶原蛋白
羟脯氨酸
Basil polysaccharide
idiopathic pulmonary fibrosis
elcithelial-mesenchymal transition
type I collagen
hydroxyproline
