Wang Dan-Dan, Wu Xin-Yue, Dong Ji-Yang, Cheng Xiu-Ping, Gu Shao-Fei, Olatunji Opeyemi Joshua, Li Yan, Zuo Jian
Xin'an Medicine Research Center, The First Affiliated Hospital of Wannan Medical College (Yijishan Hospital), Wuhu, 241000, People's Republic of China.
Research Center of Integration of Traditional Chinese and Western Medicine, Wannan Medical College, Wuhu, 241000, People's Republic of China.
J Inflamm Res. 2021 Dec 30;14:7467-7486. doi: 10.2147/JIR.S346365. eCollection 2021.
Qing-Luo-Yin (QLY) is an anti-rheumatic herbal formula. Despite the well-investigated therapeutic efficacy of QLY, its immune regulatory properties are largely unknown. CD4 T cells and monocytes are two key parameters in rheumatoid arthritis (RA). This study investigated the changes in these cells in QLY-treated RA animal models.
RA models were induced in male SD rats and were orally treated with QLY. Dynamic metabolic changes in collagen-induced arthritis (CIA) rats were monitored by H NMR approach. The immunity profiles of CIA and adjuvant-induced arthritis (AIA) rats were evaluated using immunohistochemical, PCR, ELISA, cytokine chip, flow cytometry, and immunofluorescence experiments. The bioactive components in QLY were identified by bioinformatic-guided LC-MS analyses. The compounds with high abundance in QLY decoction and easily absorbed were taken as key anti-rheumatic components and used to treat blood-derived immune cells using in vitro experiments.
The results indicated that QLY decreased Th17 cells frequency and T cells-released IL-6, IL-17 and GM-CSF in CIA rats, which was attributed to the impaired lymphocyte maturation and altered differentiation. QLY inhibited lactic acid production and inflammatory polarization in the monocytes during the peak period of AIA and CIA. AIA monocytes elicited significant increase in Th17 cells counts, IL-6 and IL-1β secretion in co-cultured splenocytes, which was abrogated by QLY. QLY-containing serum suppressed the phosphorylation of JNK and p65 in AIA lymphocyte-stimulated normal monocytes and consequently inhibited iNOS and IL-1β expression as well as IL-6 and IL-1β production. Matrine, sinomenine and sophocarpine were identified as major bioactive compounds in QLY. These identified compounds effectively inhibited the development of inflammatory T cells using concentrations detected in QLY-treated rats. At higher concentrations (20-fold increase), the chemical stimuli significantly suppressed the production of IL-1β in AIA monocytes by inhibiting JNK and p65 pathways.
By targeting inflammatory T cells and monocytes as well as disrupting their interplay, QLY improved immune environment in RA models especially during the active stages of disease.
清络饮（QLY）是一种抗风湿草药配方。尽管对QLY的治疗效果进行了充分研究，但其免疫调节特性在很大程度上仍不清楚。CD4 T细胞和单核细胞是类风湿性关节炎（RA）的两个关键参数。本研究调查了QLY治疗的RA动物模型中这些细胞的变化。
在雄性SD大鼠中诱导建立RA模型，并给予QLY口服治疗。采用核磁共振氢谱（H NMR）方法监测胶原诱导性关节炎（CIA）大鼠的动态代谢变化。使用免疫组织化学、聚合酶链反应（PCR）、酶联免疫吸附测定（ELISA）、细胞因子芯片、流式细胞术和免疫荧光实验评估CIA和佐剂诱导性关节炎（AIA）大鼠的免疫谱。通过生物信息学引导的液相色谱-质谱联用（LC-MS）分析鉴定QLY中的生物活性成分。将QLY汤剂中丰度高且易于吸收的化合物作为关键抗风湿成分，并用于体外实验中处理血液来源的免疫细胞。
结果表明，QLY降低了CIA大鼠中Th17细胞频率以及T细胞释放的白细胞介素-6（IL-6）、白细胞介素-17（IL-17）和粒细胞-巨噬细胞集落刺激因子（GM-CSF），这归因于淋巴细胞成熟受损和分化改变。在AIA和CIA的高峰期，QLY抑制了单核细胞中的乳酸产生和炎症极化。AIA单核细胞在共培养的脾细胞中引起Th17细胞计数、IL-6和IL-1β分泌显著增加，而QLY可消除这种增加。含QLY的血清抑制了AIA淋巴细胞刺激的正常单核细胞中JNK和p65的磷酸化，从而抑制了诱导型一氧化氮合酶（iNOS）和IL-1β的表达以及IL-6和IL-1β的产生。苦参碱、青藤碱和槐果碱被鉴定为QLY中的主要生物活性化合物。这些鉴定出的化合物使用在QLY治疗大鼠中检测到的浓度有效地抑制了炎性T细胞的发育。在更高浓度（增加20倍）下，化学刺激通过抑制JNK和p65途径显著抑制了AIA单核细胞中IL-1β的产生。
通过靶向炎性T细胞和单核细胞并破坏它们之间的相互作用，QLY改善了RA模型中的免疫环境，尤其是在疾病的活跃阶段。
