Tetrandrine Ameliorates Rheumatoid Arthritis in Mice by Alleviating Neutrophil Activities

Abstract

Rheumatoid arthritis (RA) is a common autoimmune disease worldwide. Neutrophils play critical roles in the onset and development of RA and are the promising target for RA treatment. Tetrandrine is a bis-benzyl isoquinoline alkaloid derived from the traditional Chinese herbal Stephania tetrandra S. Moore. Tetrandrine is effective in alleviating RA by inhibiting macrophage inflammatory response, fibroblast overproliferation, and pannus formation. However, whether tetrandrine regulates the activities of neutrophils in RA is largely unknown. In this study, we adopted adjuvant-induced arthritis (AA) murine model to explore the effect of tetrandrine on RA and neutrophils. Twenty-eight mice were divided into four groups. The control group was injected with PBS in the limbs and treated with PBS by intraperitoneal injection (i.p.) from Day 10 to Day 37. The arthritis murine model was induced by injecting FCA into the ankle joints of hind limbs. The AA group, the AA + TET group, and the AA + DEX group mice were treated with PBS, tetrandrine (6 mg/kg), or dexamethasone (1 mg/kg) i.p. daily, respectively. Arthritic scores were evaluated, and the joint diameter was measured every three days. A cytometric bead assay was performed to measure the concentrations of IFN-γ, TNF-α, and IL-6 in the serum. H&E staining and Safranin O-fast staining were adopted to monitor the tissue changes in the joint. Immunohistochemistry assays were applied to detect the MPO, NE, CitH3, and PAD4 expression levels. To assess the effect of tetrandrine on neutrophil activities in vitro, CCK8 tests were applied to determine cell viability. The qPCR and ELISA were performed to determine IL-1β and IL-6 expression levels. Immunofluorescence assays were performed to measure the formation of NETs. The results indicated that tetrandrine significantly alleviated the symptoms of RA in terms of the ankle diameter (from 4.629 ± 2.729 to 3.957 ± 0.257; P < 0.01 ) and ankle score (from 4.000 ± 0.000 to 3.286 ± 0.756; P < 0.05 ). Tetrandrine treatment significantly increased the cartilage areas and decreased serum IL-6 significantly (from 5.954 ± 2.127 to 2.882 ± 2.013; P < 0.01 ). The immunohistochemistry assays also showed decreased expression levels of NE, MPO, PAD4, and CitH3 induced by tetrandrine in comparison with the AA group ( P < 0.01 ). The qPCR assays and ELISAs showed that tetrandrine had an anti-inflammatory effect in vitro by significantly inhibiting IL-6 ( P < 0.01 ). The immunofluorescence assays showed that NET formation induced by PMA could be reduced by tetrandrine ( P < 0.01 ). In conclusion, tetrandrine has good efficacy in treating RA by regulating neutrophil-involved inflammation and NET formation.

Natural medicines of targeted rheumatoid arthritis and its action mechanism

“…Studies ( 85 , 86 ) have shown that the anti-inflammatory mechanism of tetrandrine in the treatment of RA may be related to its efficacy in inhibiting the LPS-induced phosphorylation of IκBα and NF-κB p65 in RAW 264.7 macrophages and ATDC5 chondrocytes and reducing the expression of COX in peripheral blood mononuclear cell of CFA rats. A recent study ( 87 ) has also shown that tetrandrine can improve RA by regulating neutrophil activity in AIA mice and reducing the formation of neutrophil extracellular traps induced by phorbol ester. AhR is essential for the differentiation and activation of Th17 cells ( 88 ).…”

A Worldwide Bibliometric Analysis of Tetrandrine Research in Recent Two Decades

“…3)TET for arthritis treatment: TET has been found to have an anti-arthritis impact in a great number of latest research, its anti-arthritis mechanism is visible in a variety of pathway. Liu QY demonstrated that TET reduces neutrophil activity in mice rheumatoid arthritis model ( Lu et al, 2022 ). Zhong ZY discovered that TET protects 0variectomized mice from bone Loss by inhibiting RANKL-Induced Osteoclastogenesis.…”

The bone-protective mechanisms of active components from TCM drugs in rheumatoid arthritis treatment

“…In both primary FLS isolated from the synovium of RA patients and the cell line MH7A, tetrandrine (0.3–1 μM) greatly impeded the migration and invasion of RA-FLS in vitro , as well as the expression of MMP-2/9 ( Lv et al, 2015 ). In addition, NET formation was proven to be inhibited in AA mice after tetrandrine treatment with intraperitoneal injection (6 mg/kg) ( Lu et al, 2022 ). In vitro , delineation of the reduced expression of PAD-4 and citrullinated histone H3 after tetrandrine treatment (10 μM) supported former observation in vivo ( Lu et al, 2022 ).…”

“…In addition, NET formation was proven to be inhibited in AA mice after tetrandrine treatment with intraperitoneal injection (6 mg/kg) ( Lu et al, 2022 ). In vitro , delineation of the reduced expression of PAD-4 and citrullinated histone H3 after tetrandrine treatment (10 μM) supported former observation in vivo ( Lu et al, 2022 ).…”

“…7 Jia et al (2018);Jia et al (2019); BMDMsJia et al (2018);Jia et al (2019);EL-4 Yuan et al (2016); Th17Yuan et al (2016);Yuan et al (2017); Treg Yuan et al (2016);MH7A Lv et al (2015);FLS Lv et al (2015); NeutrophilLu et al (2022) CIAYuan et al (2016); Jia et al (2018); AA Lu et al (2022) Triptolide FLS Liu et al (2013); Yang et al (2016); Piao et al (2021); PBMCs Liu et al (2013); Treg Xu et al (2016); BMDMs Xu et al (2016); Th17 Shen et al (2022); Neutrophil Huang et al (2018) CIA Liu et al (2013); Yang et al (2016); Piao et al (2021); Shen et al (2022); SCID with coimplantation Yang et al (2016); AA Huang et al (2018) Total glucosides of paeony GC B cells Li et al (2019a); Tfh cells Li et al (2019a); FLS Jia et al (2014); PBMCs Peng et al (2019); Th1 Peng et al (2019); Th2 Peng et al (2019); Th17 Peng et al (2019); Treg Peng et al (2019) CIA Jia et al (2014); Li et al (2019a); Peng et al (2019); AA Wei et al(2013)Frontiers in Pharmacology frontiersin.org effect of artesunate on the activity of osteoclast and osteoblast in vivo is worth performing.…”