Park Doil, Ko Hyun Min, Jee Wona, Park So Mi, Park Ye Rin, Jung Ji Hoon, Kim Hyung Suk, Chung Won Seok, Kim Sang Ki, Chung Jong Sup, Jang Hyeung Jin
College of Korean Medicine, Kyung Hee University, 24 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Republic of Korea.
Department of Science in Korean Medicine, Graduate School, Kyung Hee University, Seoul 02447, Republic of Korea.
Life (Basel). 2023 Feb 20;13(2):595. doi: 10.3390/life13020595.
Inflammation causes a protective immune response, which can be observed by examining the inflammatory responses of macrophages. Macrophages release various immunostimulatory factors when destroying external pathogens. We induced lipopolysaccharides (LPS) in RAW 264.7 cells, a macrophage cell line, to determine whether Helixor-M can cause immuno-suppression. Helixor-M is known to have anticancer and immune effects. However, an indicator that regulates immunity has not been clearly confirmed. To this end, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was conducted to confirm Helixor-M was not cytotoxic. Western blotting and real-time polymerase chain reaction (RT-PCR) confirmed the anti-inflammatory effects. Additionally, immunofluorescence assay confirmed the translocation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65, a representative inflammatory pathway. Helixor-M was found to be non-cytotoxic, induce the NF-κB pathway, and reduce the levels of pro-inflammatory cytokine and mitogen-activated protein kinase (MAPK). We found Helixor-M affected the PI3K/AKT/JNK pathway. Therefore, we confirmed Helixor-M acts as an anti-inflammatory agent through NF-κB, TLR4 and PI3K inhibition and that it could be an effective immunosuppressive drug.
炎症引发保护性免疫反应，这可通过检测巨噬细胞的炎症反应来观察。巨噬细胞在破坏外部病原体时会释放各种免疫刺激因子。我们在巨噬细胞系RAW 264.7细胞中诱导脂多糖（LPS），以确定Helixor-M是否会引起免疫抑制。已知Helixor-M具有抗癌和免疫作用。然而，尚未明确确认一种调节免疫的指标。为此，进行了3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）试验，以确认Helixor-M无细胞毒性。蛋白质免疫印迹法和实时聚合酶链反应（RT-PCR）证实了其抗炎作用。此外，免疫荧光试验证实了活化B细胞核因子κB（NF-κB）p65（一种代表性炎症途径）的易位。结果发现Helixor-M无细胞毒性，可诱导NF-κB途径，并降低促炎细胞因子和丝裂原活化蛋白激酶（MAPK）的水平。我们发现Helixor-M影响PI3K/AKT/JNK途径。因此，我们证实Helixor-M通过抑制NF-κB、TLR4和PI3K发挥抗炎剂的作用，并且它可能是一种有效的免疫抑制药物。
