三七总皂苷对巨噬细胞分泌NO、TGF-β_1、MMP-9的影响研究  

Study on the effect of Panax Notoginseng Saponins on The expression of NO,TGF-β_1,MMP-9 in macrophage cells

作　　者：祁燕[1] 袁嘉丽[2] 万春平[1] 赵文娟[2] 陈一纾[2] 邢海晶[2] 

机构地区：[1]云南中医学院第一附属医院,云南昆明650000 [2]云南中医学院,云南昆明650500

出　　处：《现代中西医结合杂志》2015年第23期2509-2512,共4页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：云南省教育厅科学研究基金项目(2012C033);云南白药;云南中医学院中医药创新基金项目(YB2011J08);国家自然科学基金资助项目(81460684);江苏省应用基础研究重点项目(2013FA041)

摘　　要：目的研究三七总皂苷对脂多糖（LPS）诱导RAW264.7巨噬细胞分泌一氧化氮（NO）、转化生长因子-β1（TGF-β1）、基质金属蛋白酶-9（MMP-9）及TGF-β1、MMP-9 mRNA表达的影响,在细胞水平探讨三七总皂苷治疗慢性阻塞性肺疾病（COPD）发病过程中气道炎症及气道重建的可能作用机制。方法以LPS激活巨噬细胞构建炎症细胞模型。采用MTT法测定不同浓度三七总皂苷对巨噬细胞活力的影响。分别用10,30,100,300μg/m L浓度三七总皂苷与各组细胞共同孵育,Griess法测定NO释放量,ELISA法检测细胞培养液中TGF-β1、MMP-9的含量,实时荧光定量法检测巨噬细胞TGF-β1、MMP-9 mRNA的表达量。结果 10～300μg/m L的三七总皂苷作用于巨噬细胞24 h后,细胞活性无显著变化;100μg/m L的三七总皂苷可显著抑制LPS诱导巨噬细胞NO、TGF-β1的分泌（P〈0.01或P〈0.05）,明显下调TGF-β1mRNA表达（P〈0.05）;300μg/m L的三七总皂苷可显著抑制LPS诱导巨噬细胞NO、TGF-β1、MMP-9的分泌（P〈0.01或P〈0.05）,明显下调MMP-9 mRNA表达（P〈0.05）。结论三七总皂苷在10～300μg/m L范围内对巨噬细胞无毒性作用,且可抑制巨噬细胞NO、TGF-β1、MMP-9的分泌,下调TGF-β1、MMP-9 mRNA的表达。Objective It is to investigate the effect of Panax Notoginseng Saponins on the Lipopolysaccharide-induced production of Nitric Oxide,TGF-β1,MMP-9,and on the expression of TGF-β1,MMP-9 in RAW264. 7 cell,and discuss its possible mechanism of airway inflammation and airway remodelling at the cellular level in treating with COPD. Methods Inflammatory cell model was constructed by LPS activating macrophages. The cytotoxic activity of different concentration Panax Notoginseng Saponins on macrophages was examined by using MTT methods. Inflammatory cell models were treated with Panax Notoginseng Saponins of different concentration（ 10,30,100,300 μg / m L）,the production of NO was determined by the Griess test,the TGF-β1,MMP-9 secretion in supernatants of cultured macrophages were measured by ELISA method,and the mRNA expression of TGF-β1,MMP-9 were determined by Real-Time RT-PCR. Results There was no cytotoxic on macrophages in concentration ranges（ 10-300 μg / m L） of Panax Notoginseng Saponins compared with normal control after 24 hours;Panax Notoginseng Saponins（ 100 μg / m L） significantly inhibited LPS-induced production of NO,TGF-β1,decreased the mRNA expression of TGF-β1; In addition,Panax Notoginseng Saponins（ 300 μg/m L） also inhibited LPS-induced synthesis and secretion of NO,TGF-β1and MMP-9,decreased the mRNA expression of MMP-9. Conclusion There is no cytotoxic on macrophages in concentration ranges（ 10-300 μg / m L） of Panax Notoginseng Saponins Panax and it can inhibit macrophages production of NO,TGF-β1,MMP-9 in,decreased the mRNA expression of TGF-β1,MMP-9 in macrophage cells.

关 键 词：三七总皂苷 巨噬细胞 TGF-Β1 MMP-9 NO 慢性阻塞性肺疾病 

分 类 号：R33[医药卫生—人体生理学]