Inhibitory effect of baicalin on autophagy of synovial RSC-364 cells of rats induced by lipopolysaccharide / 吉林大学学报(医学版)

Objective: To investigate the effect of baicalin on the autophagy of the synovial RSC-364 cells of the rats induced by lipopolysaccharide (LPS) through Pl3k/Akt/mTOR signal pathway, and to clarify its mechanism. Methods: The rat synovial RSC-364 cells in logarithmic growth phase were divided into control group, model group. dexamethasone (DXMS) group. 10/imol • L 1 baicalin group. 20/imol • L 1 baicalin group and 40/imol • L baicalin group. The RSC-364 cells in control group were only supplemented with culture medium, and the RSC-364 cells in the other groups were stimulated with 1 mg • L 1 LPS for 12 h to make the inflammatory cell models. The survival rates of RSC-364 cells were detected by MTT assay, and the expression levels of P13k. Akt, mTOR. Beclinl. and LC3-II mRNA in the RSC-364 cells in various groups were detected by RT-PCR method; Western blotting method was used to detect the expression levels of Beclinl. Atg5. Atg7. Atgl2. microtubule-associated protein-light chain 3-II (LC3-II ). and P62 proteins in the RSC-364 cells in various groups. Results: Compared with control group, the survival rate of RSC-364 cells in model group was significantly increased ( P-'-CO. 01). the expression levels of P13k. Akt. mTOR mRNA and P62 protein in the RSC-364 cells in model group were significantly decreased (P<0. 05 or P<0. 01). and the expression levels of Atg5. Atg7. Atgl2. LC3-II . Beclinl mRNA and proteins were significantly increased ( P<0. 01) ; compared with model group, the survival rates of RSC-364 cells in different concentrations of baicalin groups were significantly decreased (P<0. 05 or P<0. 01). the expression levels of P13k. Akt. mTOR mRNA and P62 protein in the RSC-364 cells in different concentrations of baicalin groups were significantly increased (P-'-CO. 05 or P<0. 01). and the expression levels of Atg5. Atg7. Atgl2. LC3-II and Beclinl mRNA and proteins were decreased ( P<0. 05 or P<0. 01). Conclusion: Baicalin may inhibit the LPS-induced autophagy of the RSC-364 cells by activating the Pl3k/Akt/mTOR signaling pathway.

Inhibition of C5a prevents IL-1ß-induced alternations in rat synoviocytes in vitro.

C5a is an important pro-inflammatory peptide involved in complement activation, membrane attack complex formation, immune cell chemotaxis, and allergic responses. Osteoarthritis is a disease characterized by degenerative changes in articular cartilage. It has recently been found that inflammatory responses play an important role in the pathogenesis of osteoarthritis and also in rheumatoid arthritis, where dysfunctional synoviocytes are involved. We performed a series of studies to verify our hypothesis that inhibition of C5a would prevent IL-1ß-induced alternations in rat synoviocytes. In vitro studies were performed with RSC-364 cells to examine the role of C5a in the function of synoviocytes. RSC-364 cells (a rat derived synovial cell line) were treated with IL-1ß, IL-1ß+siC5a, IL-1ß+PMX205 that is antagonist of C5aR, or left untreated. Cell cycle, proliferation, apoptosis, invasion, as well as levels of C5a, IL-17A and TNF-α expression were evaluated. We found that IL-1ß could significantly increase the proliferation and invasion capabilities of RSC-364 cells, as well as of C5a IL-17A and TNF-α expression. In contrast, inhibition of C5a by siRNA or application of antagonist of C5aR PMX205 reversed the IL-1ß-induced changes in C5a expression, cell cycle, proliferation, apoptosis, invasion, and cytokines releases. Taken together, our study results suggest that IL-1ß can increase C5a expression in RSC-364 cells, and that C5a exerts a proinflammatory effect in RSC-364 cells. Inhibition of C5a might represent a new strategy for treating rheumatoid arthritis.

Tripterygium glycosides inhibit inflammatory mediators in the rat synovial RSC-364 cell line stimulated with interleukin-1ß.

Tripterygium glycosides (TG) are extracted from a traditional Chinese medicinal herb. Using the compound, progress has been made in the treatment of rheumatoid arthritis (RA), but the underlying mechanism of its action is poorly understood. The purpose of the present study was to investigate the role of TG in preventing inflammatory arthritis. An inflammatory cell model was established in the rat synovial RSC-364 cell line via induction with interleukin (IL)-1ß. The expression of IL-32 and matrix metalloproteinases (MMP-1 and MMP-9) was determined using an enzyme-linked immunosorbent assay. Compared with the control group (without IL-1ß), IL-1ß in the treatment group induced the expression of IL-32, MMP-1 and MMP-9 in RSC-364 cells. When a different dose of TG was added to RSC-364 cells stimulated with IL-1ß, TG decreased the expression levels of IL-32, MMP-1 and MMP-9 in a dose-dependent manner. These results indicated that TG suppressed the inflammation response in RSC-364 cells. Taken together, these findings may contribute to a better understanding of the role of TG in the anti-inflammatory therapeutics for RA.

Effects of Total Saponins from Rhizoma Dioscreae Nipponicae on VEGF and AP-1 in Rat Synovial Cell Strain / 医药导报

Objective To study the effects of medicated serum with total saponins from Rhizoma Dioscreae Nipponicae (RDN) on VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α. To investigate the mechanism about total saponin from RDN inhibition of angiogenesis. Methods Medicated serum of total saponins from RDN and tripterygium (positive control) were prepared. Rat synovial cells RSC-364 were divided into four groups: the blank control,IL-17+TNF-α model,tripterygium medicated serum,and total saponins medicated serum groups. After one hour of incubation,all groups except for the blank control were incubated with both IL-17(10 μg·L-1 ) and TNF-α(10 μg·L-1 ) for 24 hours. VEGF mRNA expression in RSC-364 was detected by PrimeScriptTM real-time quantitative PCR (RT-PCR) detection kit,and the AP-1 DNA-binding activity was detected by electrophoretic mobility shift assay (EMSA). Results Compared with the control blank group,both of the VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α increased remarkably (P<0. 05,P<0.01). The VEGF mRNA expression and AP-1 activity in tripterygium medicated serum group and total saponins medicated serum group were remarkably lower than those of the model control group (P<0.05). There was no significant difference between the two medicated serum groups. Conclusion Serum medicated with total saponins from RDN can remarkably decrease VEGF mRNA expression and AP-1 activity,indicating that the total saponins from RDN could influence VEGF secretion by inhibiting the AP-1 signal transduction pathway,VEGF is the key factor of angiogenesis,thereby to restrain angiogenesis.