CAJ | 학술논문

目的探讨虫草酸和虫草素对肝星状细胞(HSC)的炎症表型和纤维化发生特性的影响.方法以内毒素(LPS,100 ng/ml)刺激传代培养的小鼠HSC株JS1的炎症反应,药物处理组在LPS刺激同时用不同浓度(10、50或200 μmol/L)的虫草酸和虫草素处理,采用实时定量逆转录PCR法检测炎症趋化因子单核细胞趋化蛋白-1(MCP-1)mRNA的表达；酶联免疫吸附法检测细胞培养上清液中MCP-1蛋白的分泌.用转化生长因子p 1(TGF β 1,10 ng/ml)刺激HSC纤维化反应,药物处理组在TGF β1刺激同时用不同浓度虫草酸和虫草素药物处理,采用Western blot法检测I型胶原和α-平滑肌肌动蛋白的表达.采用双侧t检验作统计学分析. 结果 200 μ mol/L虫草酸和虫草素处理可显著抑制LPS刺激下JS1细胞MCP-1的mRNA和蛋白表达的增加,[mRNA相对表达量:(2.07±0.29)比(3.35±0.26),t=15.90,(1.15±0.23)比(4.17±0.61),t=8.93；蛋白表达量:(1.88±0.06)比(2.33±0.06),t=10.39,(1.47±0.25)比(1.97±0.04),t=4.6,P值均＜0.05]；此外各不同浓度药物组可抑制TGF β1刺激下JS1细胞的Ⅰ型胶原和α-平滑肌肌动蛋白蛋白表达的上调,以200μmol/L药物组抑制作用最为明显. 结论虫草酸和虫草素可减轻LPS刺激下的HSC的炎症表型以及TGF β1刺激下HSC的纤维化反应,这可能是其对肝纤维化治疗作用的主要机制.
목적 탐토충초산화충초소대간성상세포(HSC)적염증표형화섬유화발생특성적영향.방법 이내독소(LPS,100 ng/ml)자격전대배양적소서HSC주JS1적염증반응,약물처리조재LPS자격동시용불동농도(10、50혹200 μmol/L)적충초산화충초소처리,채용실시정량역전록PCR법검측염증추화인자단핵세포추화단백-1(MCP-1)mRNA적표체；매련면역흡부법검측세포배양상청액중MCP-1단백적분비.용전화생장인자p 1(TGF β 1,10 ng/ml)자격HSC섬유화반응,약물처리조재TGF β1자격동시용불동농도충초산화충초소약물처리,채용Western blot법검측I형효원화α-평활기기동단백적표체.채용쌍측t검험작통계학분석. 결과 200 μ mol/L충초산화충초소처리가현저억제LPS자격하JS1세포MCP-1적mRNA화단백표체적증가,[mRNA상대표체량:(2.07±0.29)비(3.35±0.26),t=15.90,(1.15±0.23)비(4.17±0.61),t=8.93；단백표체량:(1.88±0.06)비(2.33±0.06),t=10.39,(1.47±0.25)비(1.97±0.04),t=4.6,P치균＜0.05]；차외각불동농도약물조가억제TGF β1자격하JS1세포적Ⅰ형효원화α-평활기기동단백단백표체적상조,이200μmol/L약물조억제작용최위명현. 결론 충초산화충초소가감경LPS자격하적HSC적염증표형이급TGF β1자격하HSC적섬유화반응,저가능시기대간섬유화치료작용적주요궤제.
Objective To investigate the effects of cordyceps acid and cordycepin on the inflammatory phenotype and fibrogenic property of hepatic stellate cells (HSCs).Methods An immortalized mouse HSC line (JS1) was stimulated with lippolysaccharide (LPS; 100 ng/ml) to induce an inflammatory response with or without co-administration of cordyceps acid or cordycepin in various concentrations (10,50,or 200 μmol/L).Effects of the treatments on the chemokine monocyte chemotactic protein-1 (MCP-1) mRNA expression in the cells and the protein secretion in the cell culture supematants were determined by reverse transcription and realtime quantitative PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA),respectively.In addition,JS1 cells were treated with transforming growth factor-β1 (TGFβ1; 10 ng/ml) to induce a fibrogenic response with or without co-administration of cordyceps acid or cordycepin in various concentrations (10,50,or 200 μmol/L).Effects on the expression of fibrogenic proteins including collagen type I and a-smooth muscle actin (α-SMA),were investigated by Western blot.Results High-concentration (200 μmol/L) treatments of both cordyceps acid and cordycepin significantly inhibited the LPS-induced up-regulation of MCP-1 transcription and secretion (mRNA:2.07 ± 0.29 vs.3.35 ± 0.26,t =15.90 and 1.15 ± 0.23 vs.4.17 ± 0.61,t =8.93; protein:1.88 ± 0.06 vs.2.33 ± 0.06,t =10.39 and 1.47 ± 0.25 vs.1.97 ± 0.04,t =4.60; allP ＜ 0.05).All concentrations of cordyceps acid and cordycepin inhibited the TGFβ1-induced up-regulation of collagen type Ⅰ and α-SMA protein expression.However,the effects were more robust with the 200 μmol/L concentrations (P ＜ 0.05).Conclusion Cordyceps acid and cordycepin ameliorate the LPS-induced inflammatory phenotype and TGFβ1-induced fibrogenic response of cultured HSCs.These effects may contribute significantly to the drugs' therapeutic mechanisms to inhibit and resolve liver fibrosis.