Dai Qiaomei, Li Ji, Yun Yu, Wang Jianwei
Department of Pathology, Heilongjiang University of Chinese Medicine, Harbin, China.
Department of Chinese Formulae, Heilongjiang University of Chinese Medicine, Harbin, China.
Evid Based Complement Alternat Med. 2018 Dec 16;2018:2428546. doi: 10.1155/2018/2428546. eCollection 2018.
To investigate the effect of shikonin on (CIA) collagen-induced arthritis and its influence and mechanism on the balance between Th17 cells and Treg cells.
Three doses of shikonin were administered orally to mice before the onset of CIA, and celecoxib was used as positive control drug. The arthritis response was monitored visually by macroscopic scoring and hindpaw swelling. Histology of knee was used to assess the occurrence of cartilage destruction and bone erosion. Serum collagen type II (C II) antibody levels associated with CIA were assessed with ELISAs. RT-PCR and quantitative PCR were employed to determine the mRNA expression of cytokines and TLRs in the surface of DCs in the patella with adjacent synovium and spleen in CIA. The expression of cytokines and transcription factors in the peripheral immune organs was tested by Western blotting.
Shikonin treatment suppressed the macroscopic score and incidence of arthritis. Swelling of hind paws, cartilage destruction, and serum anti-C II concentration were delayed with shikonin when compared to controls. Shikonin treatment suppressed the arthritis in a dose-dependent manner. Moreover, the expression of Th17 cytokines (IL-17A) was greatly inhibited both in the synovium and spleen in treated groups compared with those in control groups. The mRNA and protein levels of IL-10 and TGF-, however, were upregulated after shikonin treatment. The expression of Foxp3 in the synovium and spleen was upregulated, and the expression of ROR-t in the synovium and spleen was downregulated after shikonin treatment through RT-PCR, quantitative PCR, and Western blotting. The DCs in the spleen of shikonin-treated mice had lower expression of TLR4 and MyD88, and the expression of TLR2 and TLR9 in the spleen was not different between the two groups.
Shikonin has anti-inflammatory effects on CIA. Shikonin treatment can inhibit Th17 cytokines expression and induce Treg responses through inhibiting the activation of TLR4/MyD88 pathway.
探讨紫草素对胶原诱导性关节炎（CIA）的影响及其对Th17细胞与调节性T细胞（Treg细胞）平衡的影响及机制。
在CIA发病前对小鼠口服给予三种剂量的紫草素，以塞来昔布作为阳性对照药物。通过宏观评分和后爪肿胀情况直观监测关节炎反应。采用膝关节组织学检查评估软骨破坏和骨质侵蚀的发生情况。用酶联免疫吸附测定法（ELISA）评估与CIA相关的血清Ⅱ型胶原（CⅡ）抗体水平。采用逆转录-聚合酶链反应（RT-PCR）和定量聚合酶链反应测定CIA小鼠髌骨及其相邻滑膜和脾脏中树突状细胞（DCs）表面细胞因子和Toll样受体（TLRs）的mRNA表达。通过蛋白质免疫印迹法检测外周免疫器官中细胞因子和转录因子的表达。
紫草素治疗可降低关节炎的宏观评分和发病率。与对照组相比，紫草素可延缓后爪肿胀、软骨破坏及血清抗CⅡ浓度升高。紫草素治疗以剂量依赖方式抑制关节炎。此外，与对照组相比，治疗组滑膜和脾脏中Th17细胞因子（白细胞介素-17A，IL-17A）的表达均受到显著抑制。然而，紫草素治疗后白细胞介素-10（IL-10）和转化生长因子-β（TGF-β）的mRNA和蛋白水平上调。通过RT-PCR、定量PCR和蛋白质免疫印迹法检测发现，紫草素治疗后滑膜和脾脏中叉头框蛋白3（Foxp3）的表达上调，而维甲酸相关孤核受体t（ROR-γt）的表达下调。紫草素治疗组小鼠脾脏中DCs的Toll样受体4（TLR4）和髓样分化因子88（MyD88）表达较低，两组脾脏中TLR2和TLR9的表达无差异。
紫草素对CIA具有抗炎作用。紫草素治疗可通过抑制TLR4/MyD88通路的激活来抑制Th17细胞因子表达并诱导Treg反应。
