中药方剂仙方活命饮对胶原诱导性关节炎小鼠淋巴细胞分化及促炎细胞因子产生的调节作用

Herbal formula Xian-Fang-Huo-Ming-Yin regulates differentiation of lymphocytes and production of pro-inflammatory cytokines in collagen-induced arthritis mice.

作者信息

Li Jinyu, Wei Yi, Li Xue, Zhu Dashuai, Nie Bo, Zhou Jingwei, Lou Lixia, Dong Bin, Wu Aiming, Che Yongzhe, Chen Meng, Zhu Lingqun, Mu Mingwei, Chai Limin

机构信息

Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Haiyuncang Hutong No.5, , Dongcheng District, Beijing, China.

Department of orthopedics Rheumatology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China.

出版信息

BMC Complement Altern Med. 2017 Jan 5;17(1):12. doi: 10.1186/s12906-016-1526-x.

BACKGROUND

Xian-Fang-Huo-Ming-Yin (XFHM), a traditional herbal formula, has been used to treat sores and carbuncles for hundreds of years in Asia. Nowadays, its clinical effects in treatment of rheumatoid arthritis (RA) have been validated. In this study, we want to study its possible molecular mechanisms of regulating the differentiation of lymphocytes and production of pro-inflammatory cytokines in collagen-induced arthritis (CIA) mice for RA treatment.

METHODS

A high performance liquid chromatography-electrospray ionization/mass spectrometer (HPLC-ESI/MS) system was used to analyze the constituents of XFHM granules. An arthritics mouse model was induced by collagen and leflunomide (LEF) was used as a positive control medicine. Pathological changes at the metatarsophalangeal joint were studied through Safranin O and immunohistochemical staining. The differentiation of T, B and NK cells was examined by flow cytometry and pro-inflammatory cytokines were assayed using an Inflammation Antibody Array assay. The expression of key molecules of the nuclear factor κB (NF-κB) and Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathways in spleen were studied by western-blot analysis.

RESULTS

In our study. 21 different dominant chemical constituents were identified in XFHM. Treatment with XFHM suppressed the pathological changes in arthrosis of CIA. Additionally, XFHM down-regulated the proliferation and differentiation of CD3 T cells and CD3CD19 B cells significantly. However, XFHM had no significant effect on CD3NK1.1 NK cells. Further study showed that the production of pro-inflammatory cytokines had been suppressed by inhibiting the activation of NF-κB and JAK/STAT signaling.

CONCLUSIONS

XFHM can regulate and maintain the immunologic balance of lymphocytic immunity and inhibit the production of pro-inflammatory cytokines, thus suppressing the pathological changes of RA. Therefore, XFHM may be used as an application of traditional medicine against RA in modern complementary and alternative therapeutics.

背景

仙方活命饮（XFHM）是一种传统中药方剂，在亚洲已被用于治疗疮疡数百年。如今，其治疗类风湿关节炎（RA）的临床疗效已得到验证。在本研究中，我们旨在探讨其在胶原诱导性关节炎（CIA）小鼠中调节淋巴细胞分化和促炎细胞因子产生以治疗RA的可能分子机制。

方法

采用高效液相色谱-电喷雾电离/质谱仪（HPLC-ESI/MS）系统分析XFHM颗粒的成分。用胶原诱导建立关节炎小鼠模型，来氟米特（LEF）作为阳性对照药物。通过番红O染色和免疫组织化学染色研究跖趾关节的病理变化。采用流式细胞术检测T、B和NK细胞的分化情况，并使用炎症抗体阵列检测法测定促炎细胞因子。通过蛋白质免疫印迹分析研究脾脏中核因子κB（NF-κB）和Janus激酶/信号转导子和转录激活子（JAK/STAT）信号通路关键分子的表达。

结果

在我们的研究中，XFHM中鉴定出21种不同的主要化学成分。XFHM治疗可抑制CIA小鼠关节病变的病理变化。此外，XFHM显著下调CD3⁺T细胞和CD3⁺CD19⁺B细胞的增殖和分化。然而，XFHM对CD3⁺NK1.1⁺NK细胞无显著影响。进一步研究表明，通过抑制NF-κB和JAK/STAT信号的激活，促炎细胞因子的产生受到抑制。