CAJ | 학술논문

目的：探讨参麦开肺散对NIH/3T3成纤维细胞(FB)增殖及体外纤维化细胞模型胶原合成的影响。方法 NIH/3T3 FB常规培养后分为正常对照组、模型组和处理组,正常对照组加入含1%胎牛血清(FBS)的细胞培养液500μl,模型组加入使用含1% FBS细胞培养液配制的5 ng/mlβ型转化生长因子( TGF-β)溶液500μl,处理组加入含5 ng/ml的TGF-β+1 mg/ml的参麦开肺散混合溶液500μl,培养24 h。分别采用实时定量聚合酶链反应( PCR)、蛋白免疫印迹( Western blot)和Sircol assay检测细胞内胶原及细胞外基质( ECM)相关基因表达水平、Ⅰ型胶原表达情况以及分泌到细胞上清中胶原蛋白的含量。结果参麦开肺散显著降低胶原及其相关基因、Ⅰ型胶原蛋白的表达以及分泌到细胞上清中胶原蛋白的含量(P<0．05,P<0．01)。结论参麦开肺散可明显抑制TGF-β诱导的NIH/3T3 FB胶原的合成,其发挥抗纤维化作用可能与其抑制FB增殖及胶原合成相关。
목적：탐토삼맥개폐산대NIH/3T3성섬유세포(FB)증식급체외섬유화세포모형효원합성적영향。방법 NIH/3T3 FB상규배양후분위정상대조조、모형조화처리조,정상대조조가입함1%태우혈청(FBS)적세포배양액500μl,모형조가입사용함1% FBS세포배양액배제적5 ng/mlβ형전화생장인자( TGF-β)용액500μl,처리조가입함5 ng/ml적TGF-β+1 mg/ml적삼맥개폐산혼합용액500μl,배양24 h。분별채용실시정량취합매련반응( PCR)、단백면역인적( Western blot)화Sircol assay검측세포내효원급세포외기질( ECM)상관기인표체수평、Ⅰ형효원표체정황이급분비도세포상청중효원단백적함량。결과삼맥개폐산현저강저효원급기상관기인、Ⅰ형효원단백적표체이급분비도세포상청중효원단백적함량(P<0．05,P<0．01)。결론삼맥개폐산가명현억제TGF-β유도적NIH/3T3 FB효원적합성,기발휘항섬유화작용가능여기억제FB증식급효원합성상관。
Objective To investigate the effect of Shenmaikaifei powder on the proliferation and collagen synthe-sis of NIH/3T3 fibroblasts models in vitro. Methods NIH/3T3 fibroblasts underwent conventional culture, and then were divided into control group, model group and treatment group. The control group was added with 1% FBS cell culture fluid (500 μl);model group was added with 1% FBS cell culture fluid and 5 ng/ml transforming growth factor-β(TGF-β) solution ( 500 μl ); treatment group was added with 5 ng/ml TGF-βand 1 mg/ml Shenmaikaifei powder solution (500 μl);the culture time was 24 h. The real-time quantitative PCR (polymerase chain reaction) was used to examine the transcript expression levels of intra-cellular collagen and extracellular matrix ( ECM ) genes, and western blot was conducted to determine the protein expressions of type Ⅰ collagen, and Sircol assay was used to measure the collagen protein contents secreted to the cell supernatant. Results Shenmaikaifei powder significantly decreased the expressions of collagen and related genes, type I collagen and the content secreted to the cell supernatant (P<0. 05, P<0. 01). Conclusions Shenmaikaifei powder can inhibited the synthesis of collagen in TGF-β-induced NIH/3T3 fibroblasts, and its anti-fibrotic effect may relate to inhibition of fibroblast proliferation and collagen biosynthesis.