虫草酸和虫草素对肝星状细胞炎症及纤维化发生特性的影响

Effects of cordyceps acid and cordycepin on the inflammatory and fibrogenic response of hepatic stellate cells

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，100ng／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200μmol／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D1（TGFD1，10ng／m1）刺激HSC纤维化反应，药物处理组在TGFβ1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200umol／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TGFβ1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TGFp1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。

Objective To investigate the effects ofcordyceps 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 for without co-administration of cordyceps acid or cordycepin in various concentrations （10, 50, or 200μmol/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 real- time 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 Gt-smooth muscle actin （ct-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; all P〈 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.