OBJECTIVES: To investigate the role of the BNIP3-PI3K/Akt signaling pathway in mediating the inhibitory effect of Buyang Huanwu Decoction (BYHWT) on mitochondrial autophagy in human synovial fibroblasts from rheumatoid arthritis patients (FLS-RA) cultured under a hypoxic condition. METHODS: Forty normal Wistar rats were randomized into two groups (n=20) for daily gavage of BYHWT or distilled water for 7 days to prepare BYHWT-medicated or control sera. FLS-RA were cultured in routine condition or exposed to hypoxia (10% O₂) for 24 h wigh subsequent treatment with IL-1β, followed by treatment with diluted BYHWT-medicated serum (5%, 10% and 20%) or control serum. AnnexinV-APC/7-AAD double staining and T-AOC kit were used for detecting apoptosis and total antioxidant capacity of the cells, and the changes in ROS, ATP level, mitochondrial membrane potential and Ca²⁺ homeostasis were analyzed. The changes in mRNA and protein expressions of BNIP3, PI3K and AKT and mRNA expressions of LC3, Beclin-1 and P62 were detected using RT-qPCR and Western blotting. RESULTS: Treatment with BYHWT-medicated serum dose-dependently lowered apoptosis rate of IL-1β-induced FLS-RA with hypoxic exposure. The treatment significantly decreased T-AOC concentration, increased ROS production, autophagosome formation and ATPase levels, and lowered mitochondrial membrane potential and Ca²⁺ level in the cells. In IL-1β-induced FLS-RA with hypoxic exposure, treatment with BYHWT-medicated serum significantly increased BNIP3 protein expression, decreased the protein expressions of PI3K and AKT, increased the mRNA expressions of BNIP3 and P62, and lowered the mRNA expressions of PI3K, AKT, LC3 and Beclin-1 without significantly affecting Beclin-1 protein expression. The cells treated with 5% and 10% BYHWT-medicated serum showed no significant changes in LC3 expression. CONCLUSIONS BYHWT inhibits mitochondrial autophagy in IL-1β-induced FLS-RA with hypoxic exposure possibly by inhibiting BNIP3-mediated PI3K/AKT signaling pathway.
Drugs, Chinese Herbal/pharmacology* ; Arthritis, Rheumatoid/pathology* ; Animals ; Signal Transduction/drug effects* ; Proto-Oncogene Proteins c-akt/metabolism* ; Autophagy/drug effects* ; Humans ; Fibroblasts/cytology* ; Rats, Wistar ; Membrane Proteins/metabolism* ; Rats ; Phosphatidylinositol 3-Kinases/metabolism* ; Mitochondria/metabolism* ; Cells, Cultured ; Proto-Oncogene Proteins/metabolism* ; Apoptosis/drug effects* ; Cell Hypoxia ; Synovial Membrane/cytology* ; Male ; Mitochondrial Proteins

OBJECTIVES: To investigate the regulatory effect of LINC00837/miR-671-5p/SERPINE2 functional axis on pathological processes of fibroblast-like synovial cells (FLS) in rheumatoid arthritis (RA). METHODS: RA-FLS were transfected with a LINC00837 overexpression plasmid (pcDNA3.1-LINC00837), a LINC00837 interference plasmid (siRNA-LINC00837), or their respective negative control plasmids (pcDNA3.1-NC and siRNA-NC). Dual luciferase was used to verify the targeting relationship between LINC00837 and miR-671-5p and between miR-671-5p and SERPINE2. RT-qPCR was used to detect the expression levels of LINC00837, miR-671-5p and SERPINE2 in normal FLS or the transfected cells, whose proliferation and migration abilities were assessed using Edu assay and scratch healing assay and by detecting the expression levels of Ki-67, PCNA, E-cadherin and N-cadherin with Western blotting. The changes in cellular secretion of the inflammatory cytokines (TNF‑α, IL-17, IL-4 and IL-10) were examined using ELISA. RESULTS: Dual luciferase reporter gene assay showed that LINC00837 was capable of binding to the 3'-UTR of miR-671-5p, and the latter bound to the 3-UTR of SERPINE2 at specific binding sites between them. Compared with normal FLS, RA-FLS showed significantly increased expressions of LINC00837 and SERPINE2, lowered miR-671-5p expression and enhanced proliferation and migration abilities with increased expressions of pro-inflammatory cytokines and reduced expressions of anti-inflammatory cytokines. Transfection of RA-FLS with pcDNA-LINC00837 further enhanced cell proliferation and migration and the changes in the inflammatory cytokines, while transfection with si-LINC00837 produced the opposite changes. CONCLUSIONS RA-FLS have a LINC00837/miR-671-5p/SERPINE2 functional axis, which regulates cell proliferation, migration and secretion of inflammatory factors, and interventions targeting LINC00837 may provide a potential strategy to regulate the pathological processes in RA-FLS.
Arthritis, Rheumatoid/metabolism* ; MicroRNAs/metabolism* ; Humans ; Cell Proliferation ; Cell Movement ; Synovial Membrane/pathology* ; RNA, Long Noncoding/genetics* ; Fibroblasts/metabolism* ; Synoviocytes/metabolism* ; Cells, Cultured ; Transfection

OBJECTIVE: To investigate the effect of triptolide (TPL) on inflammatory response and migration of fibroblast like synovial cells (FLS) in rheumatoid arthritis (RA-FLS) and the mechanism of circular noncoding RNA (circRNA) 0003353 for mediating this effect. METHODS: We collected peripheral blood mononuclear cells (PBMCs) and serum samples from 50 hospitalized RA patients and 30 healthy individuals for detecting the expression of circRNA 0003353, immune and inflammatory indexes (ESR, CRP, RF, anti-CCP, IgA, IgG, IgM, C3, and C4) and DAS28 score. Cultured RA-FLS was treated with 10 ng/mL TPL and transfected with a circRNA 0003353 overexpression plasmid, and cell counting kit-8 (CCK-8) assay and Transwell assay were used to detect the changes in the viability and migration of the cells. Enzyme-linked immunosorbent assay (ELISA) was used to examine the cytokines IL-4, IL-6, and IL-17, and real-time fluorescence quantitative PCR (RT-qPCR) was performed to detect the expression of circRNA 003353; Western blotting was used to detect the expressions of p-JAK2, pSTAT3, JAK2 and STAT3 proteins in the treated cells. RESULTS: The expression of circRNA 0003353 was significantly increased in PBMCs from RA patients and showed a good performance in assisting the diagnosis of RA (AUC=90.5%, P < 0.001, 95% CI: 0.83-0.98). CircRNA 0003353 expression was positively correlated with ESR, RF and DAS28 (P < 0.05). Treatment with TPL significantly decreased the expression of circRNA 0003353, suppressed the viability and migration ability, decreased the expressions of IL-6 and IL-17, and increased the expression IL-4 in cultured RA-FLS in a time-dependent manner (P < 0.01). TNF-α stimulation of RA-FLS significantly increased the ratios of p-JAK2/JAK2 and p-STAT3/STAT3, which were obviously lowered by TPL treatment (P < 0.01). TPL-treated RA-FLS overexpressing circRNA 0003353 showed significantly increased cell viability and migration ability with decreased IL-4 expression and increased IL-6 and IL-17 expressions and ratios of p-JAK2/ JAK2 and p-STAT3/STAT3 (P < 0.01). CONCLUSION The expression of circRNA 0003353 is increased in PBMCs in RA patients and in RA-FLS. TPL treatment can regulate JAK2/STAT3 signal pathway and inhibit the inflammatory response and migration of RA-FLS through circRNA 0003353.
Arthritis, Rheumatoid/pathology* ; Cells, Cultured ; Diterpenes/pharmacology* ; Epoxy Compounds/pharmacology* ; Fibroblasts/pathology* ; Humans ; Interleukin-17/metabolism* ; Interleukin-4/metabolism* ; Interleukin-6/metabolism* ; Janus Kinase 2/metabolism* ; Leukocytes, Mononuclear/metabolism* ; Phenanthrenes/pharmacology* ; RNA, Circular/metabolism* ; STAT3 Transcription Factor/metabolism* ; Signal Transduction/drug effects* ; Synovial Membrane/pathology*

OBJECTIVESTo study the effect of Wenhua Juanbi Recipe (, WJR) on expression of receptor activator of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG), and tumor necrosis factor receptor superfamily member 14 (TNFRSF14, also known as LIGHT) in rats with collagen-induced arthritis (CIA).

METHODSCIA rats were generated by subcutaneous injection of bovine collagen type-II at the tail base. Sixty CIA rats were randomly assigned (10 animals/group) to: model, methotrexate (MTX)-treated (0.78 mg/kg body weight), and WJR-treated (22.9 g/kg) groups. Healthy normal rats (n=10) were used as the normal control. Treatments or saline were administered once daily by oral gavage. Rats were sacrifificed at day 28 post-treatment and knee synovium and peripheral blood serum were collected. Toe swelling degree and expression of RANKL, OPG, and LIGHT were determined by Western blot and immunohistochemistry.

RESULTSCompared with the normal group, toe swelling degree was signifificantly increased in the model group (P<0.01). After treatment, toe swelling degree decreased signifificantly in the WJR and MTX groups compared with the model group (P<0.01). Compared with the normal group, expression of RANKL and LIGHT were signifificantly increased and OPG signifificantly decreased in peripheral blood and synovium of the model group (P<0.01). Conversely, RANKL and LIGHT expression were signifificantly reduced and OPG increased in the WJR and MTX groups compared with the model group (P<0.01). No statistically significant difference existed between WJR and MTX groups.

CONCLUSIONWJR likely acts by reducing RANKL expression and increasing OPG expression, thus inhibiting RANKL/RANK interaction and reducing LIGHT expression, thereby inhibiting osteoclast formation/activation to block bone erosion.

Animals ; Arthritis, Experimental ; drug therapy ; metabolism ; Blotting, Western ; Cattle ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Immunohistochemistry ; Male ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Rats, Wistar ; Receptors, Tumor Necrosis Factor, Member 14 ; metabolism ; Synovial Membrane ; drug effects ; pathology

An unusual grayish brown discoloration of the synovium was found during a knee arthroscopy of a 72-year-old man. He also had similar pigmentation affecting the skin on the legs, arms, hands, and face. It was found he had been taking 400 mg of amiodarone hydrochloride daily for last 7 years. Amiodarone is known to cause a slate grey pigmentation of skin and cornea, but we believe this is the first report of amiodarone-induced pigmentation of the synovium. The arthroscopist should be aware of the possibility of drug-related synovial pigmentation and include this in differential diagnosis.
Aged ; Amiodarone/*adverse effects/therapeutic use ; Anti-Arrhythmia Agents/*adverse effects/therapeutic use ; Arrhythmias, Cardiac/complications/drug therapy ; Arthroscopy ; Diagnosis, Differential ; Humans ; Knee Joint/surgery ; Male ; Pigmentation Disorders/*chemically induced/*diagnosis ; Skin/pathology ; Synovial Membrane/*pathology

OBJECTIVETo compare the long-term outcomes of arthroscopy and non-surgical therapy in patients with knee synovial plica syndrome (SPS).

METHODSThirty-one patients with knee SPS undergoing arthroscopic surgery and 27 SPS patients undergoing conservative treatment with articular cavity injection of sodium hyaluronate (control group) were compared for visual pain score (VAS) and the Lysholm score before and after the operation. The patients were followed-up for more than 5 years, and the long-term outcomes were compared using Kaplan-Meier survival analysis.

RESULTSs VAS and the Lysholm scores in both groups were significantly lowered after treatment (P<0.05). The postoperative VAS score was significantly lower while the Lysholm score significantly higher in arthroscopic surgery group than in the control group (P<0.05). Arithmetic mean and the median time of curative effect maintenance were 49 and 43 months in arthroscopic surgery group, as compared with 33 and 29 months in the control group, respectively; the cumulative effect maintenance rate in arthroscopic surgery group was significantly higher than that in the control group (Χ2=4.933, P=4.933).

CONCLUSIONArthroscopic treatment produces better therapeutic effect on knee joint SPS and ensures longer long-term therapeutic effect maintenance than conservative treatment.

Adult ; Arthroscopy ; Female ; Follow-Up Studies ; Humans ; Hyaluronic Acid ; administration & dosage ; Injections, Intra-Articular ; Joint Diseases ; surgery ; Knee Joint ; pathology ; surgery ; Male ; Pain, Postoperative ; Postoperative Period ; Synovectomy ; Synovial Membrane ; pathology ; Treatment Outcome

OBJECTIVETo measure the synovial thickness in the bilateral wrist joints, metacarpophalangeal (MCP) joints and proximal interphalangeal (PIP) joints using high-resolution intraoperative ultrasound probe in healthy young and middle-aged volunteers to determine the normal reference ranges of synovial thickness in the joints.

METHODSThis study was conducted in 100 healthy young and middle-aged volunteers, including 50 male (mean age 36.2∓5.0 years; range 18-54 years) and 50 female (mean age 38.8∓5.5 years; range 20-56 years) individuals. The synovial thickness in the bilateral wrist joints, MCP joints and PIP joints was measured using the GE Logiq S8 18 MHz polo stick-like linear probe array.

RESULTSs No statistical significance was found in the synovial thickness of the wrist joints, MCP joints and PIP joints between the left and right hands, between young (<40 years) and middle-aged (≥40 years) subjects, or between the flexor surface and the extensor surface. In the male subjects, however, the synovial thickness in the wrist joints, MCP joints and PIP joints was significantly greater than that in female subjects. A significant difference was noted in the synovial thickness between the wrist joints, MCP joints and PIP joints.

CONCLUSIONMeasurement of the synovial thickness of the wrist joints, MCP joints and PIP joints using high-resolution intraoperative ultrasound probe is beneficial for early diagnosis and therapy of rheumatoid arthritis.

Adolescent ; Adult ; Arthritis, Rheumatoid ; diagnostic imaging ; Female ; Hand ; diagnostic imaging ; Humans ; Male ; Metacarpophalangeal Joint ; diagnostic imaging ; Middle Aged ; Synovial Membrane ; diagnostic imaging ; pathology ; Ultrasonography ; Wrist ; diagnostic imaging ; Wrist Joint ; diagnostic imaging ; Young Adult

Humans ; Osteoarthritis, Knee ; pathology ; surgery ; Risk Factors ; Synovectomy ; Synovial Membrane ; pathology

Adult ; Female ; Femoracetabular Impingement ; pathology ; Hip Joint ; pathology ; Humans ; Male ; Synovial Membrane ; pathology

OBJECTIVETo study the mechanism underlying the inhibitory effect of Qingluo Tongbi Granule (, QTG) on osteoclast differentiation in rheumatoid arthritis in rats.

METHODSFibroblast and monocyte co-culture were used to induce osteoclast differentiation in adjuvant-induced arthritic (AIA) rats. Serum containing QTG was prepared and added to the osteoclasts, and activation of the tumor necrosis factor receptor-associated factor 6/mitogen-activated protein kinase/nuclear factor of activated T cells, cytoplasmic1 (TRAF6/MAPK/NFATc1) pathways was examined.

RESULTSThe induced osteoclasts were multinucleated and stained positive for tartrate-resistant acid phosphatase (TRAP) staining. Serum containing QTG at 14.4, 7.2 or 3.6 g/kg inhibited the activation of TRAF6, extracellular regulated protein kinase (ERK)1/2, c-Jun N-terminal kinase (JNK) and p38 and decreased the percentage of cells with nuclear NFATc1 in a dose-dependent manner, the high and middle doses exhibited clear inhibitory activity (P<0.01 and P<0.05, respectively). After the addition of MAPK inhibitors, the NFATc1 expression showed no significant difference compared with the control group (P>0.05).

CONCLUSIONSSerum containing QTG could generally inhibit the TRAF6/MAPK pathways and possibly inhibit the NFATc1 pathway. In addition, QTG may regulate other signaling pathways that are related to osteoclast differentiation and maturation.

Adjuvants, Immunologic ; adverse effects ; Animals ; Arthritis, Experimental ; pathology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Coculture Techniques ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; pathology ; Male ; Monocytes ; pathology ; Osteoclasts ; cytology ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; pathology