Ethyl Caffeate (ETHYL 3,4-DIHYDROXYCINNAMATE) suppressed the differentiation of naive CD4+ T cells into Th1 in vitro. Furthermore, Ethyl Caffeate intensely blocked the transcriptional expression in interferon-γ-related signaling, including IFN-γ, T-bet, STAT1, and STAT4.

体内实验中，ETHYL CAFFEATE(ECF)处理显著减轻了胶原诱导性关节炎(CIA)的严重程度，抑制了IFN-γ和IL-6的分泌，并降低了CD11b+Gr-1+脾脏嗜中性白细胞的比例。同时，ECF处理在不明显影响Th17细胞和调节性T细胞发展的情况下，显著抑制了CD4+T细胞中IFN-γ的表达。

Shikui X、Aixue Z、Zengjun G 等人研究显示，Ethyl Caffeate 通过抑制Th1免疫应答，有效缓解胶原诱导的关节炎[J]。

The proliferation of splenocytes or T cells in response to ConA, LPS, and anti-CD3/28 was measured by CCK-8 Kit.?Briefly, BALB/c splenocyte suspension (5?×?105 cells/well) was cultured with ConA (5?μg/ml), LPS (10?μg/ml), and anti-CD3 (5?μg/ml;?145-2C11) in the presence of ECF at indicated concentrations.?The cultures were incubated for 48?h, 20?μl of CCK-8 was then added to each well before the end of culture, and OD value was read at 450?nm.?The MTT method was used to measure the cytotoxicity of the sample.?Splenocytes (5?×?105 cells/well) were cultured in triplicates in the absence or presence of ECF in a 96-well flat-bottomed plate (Costar) for 48?h.?MTT (5?mg/ml) was pulsed for 4?h prior to the end of the culture.?Upon removal of MTT/medium, 150?μl of DMSO was added to each well, and the plate was agitated on an oscillator for 5?min to dissolve the precipitate.?The assay plate was read at 570?nm using a microplate reader.

Collagen was dissolved in 0.1 M acetic acid at 4°C overnight.?Male DBA/1 mice were immunized at the tail base with 125?μg of collagen emulsified in complete Freund's adjuvant (CFA) containing Mycobacterium tuberculosis strain H37Rv.?Each mouse was then boosted with the same amount of collagen plus CFA 21 days later (taken as day 0). Starting from day 9 for 10 consecutive days, the mice were administered daily with ECF (50?mg/kg) or methotrexate (2?mg/kg)