Obacunone targets macrophage migration inhibitory factor (MIF) to impede osteoclastogenesis and alleviate ovariectomy-induced bone loss.

INTRODUCTION: Osteoporosis is the most common bone disorder where the hyperactive osteoclasts represent the leading role during the pathogenesis. Targeting hyperactive osteoclasts is currently the primary therapeutic strategy. However, concerns about the long-term efficacy and side effects of current frontline treatments persist. Alternative therapeutic agents are still needed. OBJECTIVES: Obacunone (OB) is a small molecule with a broad spectrum of biological activities, particularly antioxidant and anti-inflammatory effects. This study aims to examine OB's therapeutic potential on osteoporosis and explore the rudimentary mechanisms. METHODS: Osteoclast formation and osteoclastic resorption assays were carried out to examine OB's inhibitory effects in vitro, followed by the in-vivo studies of OB's therapeutic effects on ovariectomy-induced osteoporotic preclinical model. To further study the underlying mechanisms, mRNA sequencing and analysis were used to investigate the changes of downstream pathways. The molecular targets of OB were predicted, and in-silico docking analysis was performed. Ligand-target binding was verified by surface plasmon resonance (SPR) assay and Western Blotting assay. RESULTS: The results indicated that OB suppressed the formation of osteoclast and its resorptive function in vitro. Mechanistically, OB interacts with macrophage migration inhibitory factor (MIF) which attenuates receptor activator of nuclear factor kappa B (NF-κB) ligand (RANKL)-induced signaling pathways, including reactive oxygen species (ROS), NF-κB pathway, and mitogen-activated protein kinases (MAPKs). These effects eventually caused the diminished expression level of the master transcriptional factor of osteoclastogenesis, nuclear factor of activated T cells 1 (NFATc1), and its downstream osteoclast-specific proteins. Furthermore, our data revealed that OB alleviated estrogen deficiency-induced osteoporosis by targeting MIF and thus inhibiting hyperactive osteoclasts in vivo. CONCLUSION: These results together implicated that OB may represent as a therapeutic candidate for bone disorders caused by osteoclasts, such as osteoporosis.

Adenosine A2A Receptor Mediates Inhibition of Synovitis and Osteoclastogenesis after Electroacupuncture in Rats with Collagen-Induced Arthritis.

BACKGROUND: This study was to investigate the role of adenosine A2A receptors (A2AR) in inhibiting the effect of electroacupuncture (EA) on osteoclastogenesis in collagen-induced arthritis (CIA). METHODS: Wistar rats were divided into four groups: sham-control group, CIA-control group, CIA-EA group, and CIA-EA-SCH58261 (A2AR antagonist) group. We detected tumor necrosis factor-α (TNF-α), nuclear transcription factor-κB (NF-κB), receptor activator of NF-κB ligand (RANKL), protein kinase A (PKA), and extracellular regulatory protein kinase 1/2 (ERK1/2) in peripheral blood by ELISA. PKA, ERK1/2, and NF-κB in ankle joints were determined by western blotting. We evaluated the arthritis damage by histological examination and determined the number of osteoclasts by tartrate-resistant acid phosphatase (TRAP) staining. RESULTS: EA treatment downregulated the expression of TNF-α, RANKL, PKA, ERK1/2, and NF-κB in peripheral blood but increased the levels of PKA and ERK1/2 in ankle joints. Importantly, EA treatment reduced bone erosion as evidenced by the histological findings and inhibited osteoclastogenesis as revealed by TRAP staining. All these effects of the EA treatment were reversed by combining EA treatment with the A2AR antagonist SCH58261. CONCLUSION: Our data suggest that EA treatment activated A2AR. The effects of the A2AR antagonist SCH58261 suggest that the inhibition of osteoclast formation, the inhibition of TNF-α, RANKL, and NF-κB expression, and the increase of ERK1/2 are all dependent on this EA-induced A2AR activation. It is therefore likely that these pathways with clearly defined roles in inflammation and bone erosion are at least partially involved in the mediation of the inhibition of synovitis and osteoclast formation induced by EA.

[Clinical observation of the phased acupuncture for ischemic stroke hemiplegia].

OBJECTIVE: To compare the effect difference among the staged acupuncture combined with conventional rehabilitation, traditional acupuncture combined with conventional rehabilitation, and conventional rehabilitation in the patients with ischemic stroke hemiplegia. METHODS: A total of 145 patients with ischemic stroke hemiplegia were randomly assigned into an observation group (49 cases), a control group A (49 cases) and a control group B (47 cases). All the patients were received aspirin tablets 100 mg orally, once a day. The staged acupuncture according to Brunnstrom's theory combined with conventional rehabilitation were used in the observation group, at the acupoints in the yangming meridian during the flaccid paralysis phase, at the acupoints in the antagonistic muscle and the tendon of spastic dominant muscle during the spasm stage, and at the acupoints in the yangming meridian and Taixi (KI 3), Guanyuan (CV 4) and Qihai (CV 6) during the sequela stage. Conventional traditional acupuncture at the acupoints in the yangming meridian combined with conventional rehabilitation were applied in the control group A. Simple conventional rehabilitation was used in the control group B. Patients in the observation group and the control group A were received acupuncture and rehabilitation once a day from Monday to Friday, 2 weeks as 1 course; patients in the control group B were received the same rehabilitation as the observation group. The indexes were observed before treatment and at the end of the 2nd, 4th, 6th and 8th weeks of treatment, including National Institutes of Health Stroke Scale (NIHSS), short form Fugl-Meyer motor function assessment (FMA) scale, Fugl-Meyer balance (FM-B) scale, comprehensive spasm scale (CSS) and modified Barthel index (MBI). RESULTS: Compared with those before treatment, except the MBI score in the control group B after 2 weeks' treatment, the NIHSS, FMA, FM-B and MBI scores were improved in all three groups at the end of the 2nd, 4th, 6th and 8th weeks of treatment (P<0.01, P<0.05). At the end of the 2nd week of treatment, the NIHSS and CSS scores were lower (P<0.05, P<0.01), the FMA, FM-B and MBI scores were higher (all P<0.05) in the observation group and the control group A than those in the control group B. After 4, 6 and 8 weeks' treatment, the FMA, FM-B and MBI scores were higher (P<0.05, P<0.01), the NIHSS and CSS scores were lower (all P<0.01) in the observation group than those of control group A and control group B; the FMA, FM-B and MBI scores in the control group A were higher than those in the control group B (P<0.05, P<0.01); the NIHSS and CSS scores in the control group A were lower than those in the control group B (P<0.05, P<0.01). CONCLUSION: Staged acupuncture combined with conventional rehabilitation are consistent with the rehabilitation rule of patients with hemiplegia. They can improve motor function and the ability of daily life. They achieve better effect than traditional acupuncture combined with rehabilitation.

Transcutaneous acupoint electrical stimulation pain management after surgical abortion: A cohort study.

INTRODUCTION: Transcutaneous acupoint electrical stimulation (TEAS) is a standard therapy for painful conditions. This study evaluated pain-relieving effects of treatment with TEAS before and after surgical abortion. METHODS: In this cohort study 140 nulliparae requesting pregnancy termination with intravenous anesthesia from August to December 2013 at the outpatient clinic of Wenzhou Medical University First Affiliated Hospital were recruited and divided into three cohorts who received TEAS pre-, post-, and both pre- and post-operation, alongside a control group. The cohorts underwent TEAS treatment for 30 min before and/or after the procedure while the control group received no TEAS treatment. Pain levels were evaluated upon recovery at 10, 30, and 45 min, respectively, after abortion. RESULTS: Mean Visual Analog Scale (VAS) scores in pre-operation cohorts, but not the post-operation cohort, were significantly lower than those obtained for the control group at 10 min (p < 0.01). VAS scores at 30 min and 45 min postoperatively were similar in each cohort but lower than control values (p < 0.001). More cohort patients reported mild or no pain than control patients (p < 0.05); the pre-operation cohorts had more women with no pain compared with the post-operation group (p < 0.05). There were no differences among groups in medical treatment required after 45 min. There were fewer complications of nausea and vomiting in the cohorts compared with the control group (p < 0.05). CONCLUSIONS: Performing TEAS before and after surgical abortion provides postoperative pain relief. However, receiving TEAS before surgery allowed more women to experience mild or no pain. IMPLICATIONS: Transcutaneous acupoint electrical stimulation shows potential as an adjunct to conventional pain treatment following surgical abortion in nulliparae.

Repeated Electroacupuncture Persistently Elevates Adenosine and Ameliorates Collagen-Induced Arthritis in Rats.

The aim of this paper was to investigate the effect of repeated electroacupuncture (EA) over 21 days on the adenosine concentration in peripheral blood of rats with collagen-induced arthritis (CIA). Wistar rats were divided into three groups of 6 animals each: sham-control, CIA-control, and CIA-EA. We determined the adenosine concentration in peripheral blood and assessed pathological changes of ankle joints. Quantitative reverse-transcription-polymerase chain reaction was used to determine mRNA levels of ecto-5'-nucleotidase (CD73), adenosine deaminase (ADA), and tumor necrosis factor-alpha (TNF-α). Immunohistochemical staining was used to detect expression of ADA and CD73 in synovial tissue. Repeated EA treatment on CIA resulted in the persistence of high concentrations of adenosine in peripheral blood, significantly reduced pathological scores, TNF-α mRNA concentrations, and synovial hyperplasia. Importantly, EA treatment led to a significant increase in CD73 mRNA levels in peripheral blood but was associated with a decrease of CD73 immunostaining in synovial tissue. In addition, EA treatment resulted in a significant decrease of both ADA mRNA levels in peripheral blood and ADA immunostaining in synovial tissue. Thus, repeated EA treatment exerts an anti-inflammatory and immunoregulatory effect on CIA by increasing the concentration of adenosine. The mechanism of EA action may involve the modulation of CD73 and ADA expression levels.

Adenosine A2A Receptors Mediate Anti-Inflammatory Effects of Electroacupuncture on Synovitis in Mice with Collagen-Induced Arthritis.

To study the role of adenosine A2A receptor (A2AR) in mediating the anti-inflammatory effect of electroacupuncture (EA) on synovitis in collagen-induced arthritis (CIA), C57BL/6 mice were divided into five treatment groups: Sham-control, CIA-control, CIA-EA, CIA-SCH58261 (A2AR antagonist), and CIA-EA-SCH58261. All mice except those in the Sham-control group were immunized with collagen II for arthritis induction. EA treatment was administered using the stomach 36 and spleen 6 points, and stimulated with a continuous rectangular wave for 30 min daily. EA treatment and SCH58261 were administered daily from days 35 to 49 (n = 10). After treatment, X-ray radiography of joint bone morphology was established at day 60 and mouse blood was collected for ELISA determination of tumor necrosis factor alpha (TNF-α) levels. Mice were sacrificed and processed for histological examination of pathological changes of joint tissue, including hematoxylin-eosin staining and immunohistochemistry of A2AR expression. EA treatment resulted in significantly reduced pathological scores, TNF-α concentrations, and bone damage X-ray scores. Importantly, the anti-inflammatory and tissue-protective effect of EA treatment was reversed by coadministration of SCH58261. Thus, EA treatment exerts an anti-inflammatory effect resulting in significant protection of cartilage by activation of A2AR in the synovial tissue of CIA.

[Electroacupuncture for post-stroke urinary incontinence: a multi-center randomized controlled study].

OBJECTIVE: To evaluate the clinical efficacy of electroacupuncture(EA) for apoplectic urinary incontinence. METHODS: Two hundred and four cases of apoplectic urinary incontinence were randomized into an EA group (136 cases) and an indwelling catheter group (68 cases). The EA was applied at Qugu (CV 2), Zhongji (CV 3), Shuidao (ST 28), Qihai (CV 6) and Guanyuan (CV 4), etc. in the EA group,5 times a week. Indwelling catheter was applied in the indwelling catheter group at intervals of 2-4 hours, and periodic bladder irrigation along with bladder rehabilitation training were also given. The efficacies were evaluated after 4 weeks of treatment. Before and after treatment, the urination diary (including the interval of urination, nocturia frequency, urination difficulty, urinary incontinence severity), bladder capacity, patients' satisfaction of the two groups were observed and the efficacy was evaluated. RESULTS: The total effective rate was 96.2% (125/130) in the EA group, which was apparently superior to 87.5% (56/64) in the indwelling catheter group (P < 0.05); except for nocturia frequency in the indwelling catheter group, the total score and the subitem score in the urination diary were all improved significantly after treatment in both groups (all P < 0.001), which were more obvious in the EA group (P < 0.001, P < 0.05); the patients' satisfaction and bladder capacity were all improved significantly after treatment in both groups (all P < 0.001), which were more obvious in the EA group (both P < 0.001). CONCLUSION: The EA has an obvious effect for apoplectic urinary incontinence in urinary incontinence alleviation and bladder capacity increase, which has better efficacy than indwelling catheter therapy.