Therapeutic effects of columbianadin from Angelicae Pubescentis radix on the progression of collagen-induced rheumatoid arthritis by regulating inflammation and oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE: Angelicae pubescentis radix (APR) has a long history in the treatment of rheumatoid arthritis (RA) in China. It has the effects of dispelling wind to eliminate dampness, removing arthralgia and stopping pain in the Chinese Pharmacopeia, but its mechanisms was remained unclear. Columbianadin (CBN), one of the main bioactive compounds of APR, has many pharmacological effects including anti-inflammatory and immunosuppression. However, there are few reports on therapeutic effect of CBN on RA. AIM OF THE STUDY: A comprehensive strategy via incorporating pharmacodynamics, microbiomics, metabolomics, and multiple molecular biological methods was adopted to evaluate the therapeutic effects of CBN on collagen-induced arthritis (CIA) mice and explore the potential mechanisms. MATERIALS AND METHODS: A variety of pharmacodynamic methods were used to evaluate the therapeutic effect of CBN on CIA mice. The microbial and metabolic characteristics of CBN anti-RA were obtained by metabolomics and 16S rRNA sequencing technology. The potential mechanism of CBN anti-RA was predicted through bioinformatics network analysis, and verified by a variety of molecular biology methods. RESULTS: CBN can effectively improved symptoms of rheumatoid arthritis in CIA mice, including paw swelling and arthritic scores. The inflammatory and oxidative stress were effectively regulated by the treatment of CBN. The fecal microbial communities and serum and urine metabolic compositions were significantly altered in CIA mice, CBN can ameliorate the CIA-associated gut microbiota dysbiosis, and regulate the disturbance of serum and urine metabolome. The acute toxicity test showed that the LD50 of CBN was greater than 2000 mg kg-1. CONCLUSIONS: CBN exert anti-RA effects from four perspectives: inhibiting inflammatory response, regulating oxidative stress, and improving changes in gut microbiota and metabolites. The JAK1/STAT3, NF-κB and Keap1/Nrf2 pathway may be important mechanism for CBN's inflammatory response and oxidative stress activity. CBN could be considered as a potential anti-RA drug for further study.

Immunosuppressive effect of Columbianadin on maturation, migration, allogenic T cell stimulation and phagocytosis capacity of TNF-α induced dendritic cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Angelicae pubescentis radix (APR) has a long history in the treatment of rheumatoid arthritis (RA) in China. It has the effects of dispelling wind to eliminate dampness, removing arthralgia and stopping pain in the Chinese Pharmacopeia, but its mechanisms was unclear. Columbianadin (CBN) was one of the main bioactive compounds of APR, and has many pharmacological effects. But the immunosuppressive effect of CBN on DCs and the potential mechanism needed to be explored. AIM OF THE STUDY: The study was aimed to clarify the immunosuppressive effect of CBN on maturation, migration, allogenic T cell stimulation and phagocytosis capacity of TNF-α induced DCs. MATERIALS AND METHODS: Bone marrow-derived DCs were obtained and cultured from C57BL/6 mice in accordance with protocol. The phenotypic study (CD11c, CD40, CD80, CD86 and MHC Ⅱ) were measured by flow cytometry. FITC-dextran were uptaked by DCs and the change of endocytosis activity were mediated by acquired mannose receptor. Transwell chambers were used to detect the migration ability of DCs. Mixed leukocyte reaction (MLR) assay was used to detect the allostimulatory ability of CBN on TNF-α stimulated DCs. The secretion of cytokines and chemokines was measured by ELISA Kit. TLRs gene and MAPKs/NF-κB protein expression were checked by qRT-PCR and Western blot. RESULTS: CBN inhibited the maturation of TNF-α-induced DCs while maintaining phagocytosis capabilities. Additionally, CBN inhibited the migration of TNF-α stimulated DCs, which related to reduce the production of chemokines (MCP-1, MIP-1α). Notably, CBN could suppress the proliferation of CD4+T cells by inhibiting DCs maturation, and decrease the proinflammatory cytokines IL-6 production. Furthermore, CBN inhibited mRNA expression of TLR2, TLR7 and TLR9 in TNF-α-activated DCs. Meanwhile, the phosphorylation of p38, JNK1/2 and NF-κB protein were significantly inhibited in CBN treated DCs. CONCLUSIONS: These findings provided novel insights into the pharmacological activity of CBN. They also indicated that inhibition DCs maturation owning to the immunosuppressive effect of CBN. CBN was expected as a potential immunosuppressant and TLRs/MAPKs/NF-κB pathway may be an important mechanism for CBN's immunosuppressive activity.

Optimization of the Purifying Process for Columbianetin-ß-D-Glucopyranoside from Angelicae Pubescentis Radix and Evaluation of Its Analgesic Activity Using Hot Plate Test.

The objective of this work was to provide an economic and practical method for the purification of columbianetin-ß-D-glucopyranoside from Angelicae Pubescentis Radix extract. In the static adsorption and desorption, the effects of resin type (D101, HP-20, AB-8, GDX-201, and DA201), contact time (10-360 min), and temperature (298-318 K) were assessed on columbianetin-ß-D-glucopyranoside adsorption efficiency in laboratory. GDX-201 resin showed the best adsorption and desorption properties for columbianetin-ß-D-glucopyranoside. The kinetic data revealed that the equilibrium time for columbianetin-ß-D-glucopyranoside adsorption was achieved within 150 min. Moreover, the adsorption kinetic curve was well in accordance with the pseudo-second-order equation (R 2 > 0.99). The rate controlling step of the adsorption process was the intraparticle diffusion. The Langmuir isotherm models (R 2 > 0.99) could describe the whole adsorption process, which was exothermic and spontaneous according to the result of thermodynamics tests. In the dynamic adsorption and desorption process, the optimum loading flow (4, 5, and 6 BV/h), ethanol concentration (0-60%), and elution volume (10-230 mL) were optimized. Under optimal conditions of 4 BV/h loading flow, 6.7 BV loading volume, 25% ethanol, and 14 BV elution volume, the content of columbianetin-ß-D-glucopyranoside in the product was increased 29.61-fold from 0.45% to 13.32 ± 0.64% with yield of 88.03 ± 2.76% by an experiment of lab-scale enlargement. Then, columbianetin-ß-D-glucopyranoside was further purified by PHPLC and its purity was more than 98%. Additionally, the analgesic activity of the columbianetin-ß-D-glucopyranoside was assessed by hot plate test. The experimental results showed that columbianetin-ß-D-glucopyranoside significantly increased the latency of pain response in mice. This study demonstrated columbianetin-ß-D-glucopyranoside could be as a potentially natural analgesic component. It could be summed up that the established method was successfully applied to purifying columbianetin-ß-D-glucopyranoside from Angelicae Pubescentis Radix extract.

Traditional herbal medicine: Therapeutic potential in rheumatoid arthritis.

ETHNOPHARMACOLOGICAL RELEVANCE: Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease influenced by diverse endogenous and exogenous factors. It is characterized by cartilage and bone destruction. The current conventional allopathic therapy is expensive and carries adverse side effects. Recently, there were some ethnopharmacological studies on RA including anti-RA effects and therapeutic targets of distinct dosage forms of traditional herbal medicines (THMs). AIM OF THE REVIEW: This review provides a brief overview of the current understanding of the potential pharmacological mechanisms of THMs (active constituents, extracts and prescriptions) in RA. This study is intended to provide comprehensive information and reference for exploring new therapeutic strategies of THMs in the RA treatment. MATERIALS AND METHODS: This review captured scientific literatures invivo and vitro experiments on effects of anti-RA THMs published between 2016 and 2021 from journals and electronic databases (e.g. PubMed, Elsevier, Science Direct, Web of Science and Google Scholar). Relevant literatures were searched and analyzed by using keywords such as 'rheumatoid arthritis AND traditional herbal medicines', 'rheumatoid arthritis AND immune cells', 'rheumatoid arthritis AND inflammation', 'rheumatoid arthritis AND miRNA', 'rheumatoid arthritis AND Angiogenesis', 'rheumatoid arthritis AND oxidative stress', 'rheumatoid arthritis AND osteoclasts', 'rheumatoid arthritis AND CIA model', 'rheumatoid arthritis AND AA model' AND 'rheumatoid arthritis herbal prescription'. RESULTS: Experiments in vitro and in vivo jointly demonstrated the potential of THMs in the RA treatment. There are plentiful therapeutic targets in RA. THMs and active ingredients could alleviate RA symptoms through different therapeutic targets, such as immunoregulation, inflammation, fibroblast-like synoviocytes (FLSs), microRNAs (miRNAs), angiogenesis, oxidative stress, osteoclasts and multiple targets interaction. Anti-RA THMs, active ingredients and prescriptions through corresponding therapeutic targets were summarized and classified. CONCLUSIONS: Flavonoids, phenolic acids, alkaloids and triterpenes of THMs are identified as the main components to ameliorate RA. Regulation of different and multiple related therapeutic targets by THMs and their active ingredients were associated with greater therapeutic benefits, among which inflammation is the main therapeutic target. Nonetheless, further studies are required to unravel the complexities and in-depth mechanisms of THMs in alleviating RA.

Buyang Huanwu Decoction ameliorates atherosclerosis by regulating TGF-ß/Smad2 pathway to promote the differentiation of regulatory T cells.

ETHNOPHARMACOLOGICAL RELEVANCE: As a classic prescription of Traditional Chinese Medicine in Correction on the Errors of Medical Works, Buyang Huanwu Decoction (BYHWD) has a good curative effect on prevention of atherosclerosis (AS). AIM OF THE STUDY: This study aims to elucidate the anti-atherosclerosis mechanism of BYHWD, which may promote the differentiation of regulatory T cells by regulating the TGF-ß/Smad2 pathway. MATERIALS AND METHODS: ApoE-/- mice were fed a high-fat diet for 12 weeks, then drugs group were given BYHWD with intragastric administration once a day for 4 weeks. The effect of BYHWD on lipid content in peripheral blood and plaque was evaluated by blood lipid test and oil red O staining. The number of Tregs in peripheral blood was tested by flow cytometry, and that in the spleen was evaluated by immunohistochemistry methods. Gene and protein expression relating with Tregs differentiation pathway in mice were checked by RT-PCR and Western blot experiments. CD4+T cells were isolated and interfered by BYHWD drug-loaded serum. The proportion of Tregs was evaluated by flow cytometry. The chemical compositions of BYHWD and rat drug-loaded serum were analyzed by ultra-high performance liquid chromatograph and liquid chromatography-tandem mass spectrometry. RESULTS: BYHWD significantly reduced plaque area and cholesterol accumulation, increased the number of Tregs in spleen and peripheral blood of ApoE-/- AS mice, raised the proportion of Tregs in CD4+T cells, and regulated the levels of inflammatory factors. It also increased the TGF-ß and Smad2 mRNA and protein levels relating with Tregs differentiation pathway in vivo. The mRNA levels of Foxp3/TGF-ß/Smad2 were enhanced via BYHWD in vitro. CONCLUSIONS: BYHWD regulates TGF-ß/Smad2 signaling pathway to promotes the peripheral differentiation of Tregs, increases the number of Tregs, restores the immune balance between CD4+T cells, regulates lipid metabolism, inhibits inflammatory reaction and possesses the potential of enhancing plaque stability.