miR-124-3p improves mitochondrial function of renal tubular epithelial cells in db/db mice.

Diabetic kidney disease (DKD) is one of the most serious complications of diabetes mellitus (DM) and the main cause of end-stage renal failure. However, the pathogenesis of DKD is complicated. In this study, we found that miR-124-3p plays a key role in regulating renal mitochondrial function and explored its possible mechanism in DKD progression by performing a series of in vitro and in vivo experiments. Decreased expression of miR-124-3p was found in db/db mice compared to db/m mice. Moreover, miR-124-3p down-regulated FOXQ1 by targeting FOXQ1 mRNA 3'-UTR in NRK-52E cells. Also, an increase in FOXQ1 and down-regulation of Sirt4 were found in db/db mouse kidney and renal tubular epithelial cells cultured with high glucose and high lipid. Overexpression of FOXQ1 could further down-regulate the expression of Sirt4 and aggravate the damage of mitochondria. Conversely, the knockdown of the FOXQ1 gene induced Sirt4 expression and partially restored mitochondrial function. To verify the effects of miR-124-3p on Sirt4 and mitochondria, we found that miR-124-3p mimics could up-regulate Sirt4 and inhibit ROS production and MitoSOX, thus restoring the number and morphology of mitochondria. These results showed that under high-glucose and high-lipid conditions, the down-regulation of miR-124-3p induces FOXQ1 in renal tubular epithelial cells, which in turn suppresses Sirt4 and leads to mitochondrial dysfunction, promoting the development of DKD.

Silver Needle Thermal Therapy Relieves Pain, Repairs the Damaged Myofascial Fiber, and Reduces the Expression of 5-HT3 Receptors in the Spinal Cord of Rats with Myofascial Pain Syndrome.

Background: There is an urgent clinical need to provide a theoretical basis for silver needle thermal therapy to Myofacial pain syndrome (MPS). Objective: This study was conducted to explore the effect of silver needle thermal therapy on myofascial pain syndrome in rats. Methods: MPS rat models were duplicated, and the rats were subsequently divided into model and treatment groups. A normal control group was synchronously set up. No treatment was given to the model group, whereas silver needle thermal therapy was administered to the treatment group. The thermal and mechanical pain threshold, the morphological structure as well as the expression of 5-HT3 receptors in the spinal cord were observed. Results: Rats from the treatment group presented with a significantly higher pain threshold compared to the untreated model group.The myofascial arrangement of the affected part of the model group was disordered, and some muscle fibers were atrophied and deformed. Meanwhile, the myofascial arrangement of the treatment group became more regular than that of the model group. The expression levels of 5-HT3 receptor in the spinal cord of the untreated model group were significantly increased, while being markedly decreased in the treatment group. Conclusions: Silver needle thermal therapy can augment the pain threshold of rats with MPS, repair the damaged myofascial membrane in the rats, and further reduce the expression of 5-HT3 receptors in the spinal cord of the MPS rats.

C5a aggravates dysfunction of the articular cartilage and synovial fluid in rats with knee joint immobilization.

Degenerative alterations in articular cartilage are involved in the pathogenesis of osteoarthritis. The present study aimed to evaluate the role of complement component 5a (C5a) in osteoarthritic alterations in the articular cartilage and synovialis via a joint immobilization (IM) rat model. Rats were assigned to three groups: Control, IM and IM+anti­C5a antibody (IM+anti­C5a) groups. A terminal deoxynucleotidyl transferase dUTP nick end labeling assay and hematoxylin and eosin staining were used to evaluate the morphological alterations in the articular cartilage and synovialis. Reverse transcription­quantitative polymerase chain reaction (RT­qPCR) analysis, immunohistochemical analysis and western blotting were used to evaluate C5a expression in the articular cartilage and synovialis. An ELISA was used to evaluate C5a­induced alterations in interleukin (IL)­1ß, IL­17A and tumor necrosis factor (TNF)­α levels in the serum and joint fluid. The results demonstrated that knee joint immobilization induced destruction of knee joint synovial fluid and cartilage in the IM and IM+anti­C5a antibody groups. Immobilization significantly increased the expression levels of C5a in serum and joint fluid in the IM group. Immunohistochemistry, western blotting and RT­qPCR analysis illustrated markedly increased expression of C5a in the IM group. Immobilization markedly increased the IL­1ß, IL­17A and TNF­α expression levels in the serum and joint fluid in the IM group. Anti­C5a was able to decrease immobilization­induced alterations in morphology and cytokines compared with the IM group. The expression of C5a was increased in synoviocytes and joint cartilage in the IM model. Pro­inflammatory cytokines, including TNF­α and IL­1ß were released in the activated synoviocytes via the induction of C5a, suggesting that C5a serves an important role in joint inflammatory processes.

Ketamine attenuates osteoarthritis of the knee via modulation of inflammatory responses in a rabbit model.

The aim of the present study was to investigate the efficacy of ketamine in attenuating osteoarthritis (OA) and modulating the expression of inflammatory mediators. A rabbit OA model was established by knee immobilization using plaster bandages. After six weeks, rabbits were randomly allocated into four groups (n=6/group): Normal saline, Ket60, Ket100, and Ket200 and twice a week for four weeks the rabbits received an intra­articular injection of saline, or 60, 100 or 200 µmol/l ketamine, respectively. One week after the final injection, samples of synovial membrane, synovial fluid and articular cartilage were isolated. The pathological changes were assessed by general observation, hematoxylin and eosin staining and Alcian blue/periodic­acid Schiff staining. Cartilage pathology was assessed using Mankin's scoring system. Tumor necrosis factor (TNF)­α and interleukin (IL)­10 levels in the synovial fluid were measured by enzyme­linked immunosorbent assays. The nuclear factor (NF)­κB p65 subunit expression level in cartilage samples was determined by immunohistochemistry. OA was characterized by morphological changes in the articular surface, cartilage lesions, infiltration of inflammatory cells and a significantly increased Mankin's score. Elevated TNF­α and reduced IL-10 levels in the synovial fluid, along with increased p65 expression levels in the cartilage were observed in OA rabbits. Intra­articular injection of ketamine ameliorated the pathological characteristics of OA, reduced the Mankin's score, decreased TNF­α and NF­κB p65 expression levels, and increased the level of IL­10 expression in a dose-dependent manner. Thus is was demonstrated that Ketamine suppresses the inflammatory response in OA by modulating inflammatory mediator expression levels in a rabbit model of OA.