ABSTRACT
OBJECTIVE: To evaluate the effect of soluble total proteins of Zaocys dhumnades on the expressions of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and IL-10 in human fibroblast-like synoviocytes (FLS) cultured in vitro. METHODS: Primary cultured FLS isolated from the synovium of patients with rheumatoid arthritis (RA) were incubated in the presence of different concentrations (50, 150 and 450 microg/ml) of soluble total proteins of Zaocys dhumnades, with Tripterygium hypoglaucum Hutch (THH) and DMEM as the positive and negative controls, respectively. Enzyme-linked immunosorbent assay (ELISA) and RT-PCR were used to detect the expressions of IL-1beta, IL-10 and TNF-alpha in the FLS. RESULTS: The protein and mRNA levels of IL-1beta and TNF-alpha in the supernatant of the FLS exposed to 150 and 450 microg/ml of the soluble total proteins of Zaocys dhumnades decreased, while IL-10 protein and mRNA increased significantly as compared with those in the negative control group (P<0.01). CONCLUSION: The soluble total proteins of Zaocys dhumnades produce therapeutic effect on RA possibly by inhibiting IL-1beta and TNF-alpha and promoting IL-10 expressions in the FLS.
Subject(s)
Colubridae , Interleukin-1beta/metabolism , Materia Medica/pharmacology , Synovial Membrane/metabolism , Tumor Necrosis Factor-alpha/metabolism , Animals , Arthritis, Rheumatoid , Cells, Cultured , Humans , Interleukin-10/metabolism , Materia Medica/chemistry , Primary Cell Culture , Proteins/isolation & purification , Synovial Membrane/cytology , Synovial Membrane/pathologyABSTRACT
OBJECTIVE: To investigate the effect of the type II collagen (C II) protein from Zaocys on cytokines production by synoviocytes in rats with adjuvant arthritis (AA). METHODS: Type II collagen protein was abstracted and purificated from Zaocys. Adjuvant arthritis (AA) was induced by a single intradermal injection of 0.1 mL of complete Freund's adjuvant into the left hind paw. Synoviocytes' supernatants were harvested and synoviocytes-Peyer's Patches (PP) cells coculture system were developed. Tumor necrosis factor-alpha (TNF-alpha) activity was measured by L929 cytotoxicity bioassay and Interleukin (IL)-1beta activity was measured by MTT dye reduction. The synoviocytes' supernatants cytokines' levels were detected by ELISA. RESULTS: Each concentration of C II from Zaocys had no effect on IL-1beta and TNF-alpha production by synoviocytes in vitro. Middle concentration of C II suppressed the activity of IL-1beta and TNF-alpha production by synoviocytes-PP cells coculture system (P < 0.05). Treating with low and high dose of C II suppressed the activity of TNF-alpha and IL-1beta producing by synoviocyte (P < 0.05), significantly suppressed in the group of AA rats treated with middle dose of C II (P < 0.01). Treating with middle and high dose of C II decreased the level of synoviocytes' supernatants TNF-alpha (P < 0.05), the level of synoviocytes' supernatants IL-1beta decreased in all treating groups (P < 0.05). Treating with middle dose of C II increased the level of serum TGF-beta (P < 0.05). Middle concentration of C II suppressed the activity of IL-1 and TNF production by synoviocytes-PP cells coculture system (P < 0.05). CONCLUSIONS: C II from Zaocys has no direct effect on the activity of IL-1beta and TNF production by synoviocytes in vitro. Oral administration of type II collagen protein from Zaocys can effectively suppressed the activity and level of the cytokines production by synoviocytes in rats with adjuvant arthritis (AA).
Subject(s)
Arthritis, Experimental/drug therapy , Collagen Type II/pharmacology , Colubridae , Cytokines/metabolism , Materia Medica/pharmacology , Synovial Membrane/drug effects , Animals , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Arthritis, Experimental/chemically induced , Arthritis, Experimental/metabolism , Cells, Cultured , Collagen Type II/administration & dosage , Collagen Type II/therapeutic use , Cytokines/drug effects , Disease Models, Animal , Female , Interleukin-1/metabolism , Materia Medica/administration & dosage , Materia Medica/therapeutic use , Random Allocation , Rats , Rats, Sprague-Dawley , Synovial Membrane/metabolism , Synovial Membrane/pathology , Tumor Necrosis Factor-alpha/drug effects , Tumor Necrosis Factor-alpha/metabolismABSTRACT
OBJECTIVE: To investigate the biological effects of the serum of rats fed with kidney-tonifying traditional Chinese drugs on cultured osteoblasts in vitro. METHODS: The culture media containing the serum from rats fed with the drugs at different doses (high, mediate and low doses) were used to treat the second passage of the osteoblasts from the skull of newborn SD rats, and the cell proliferation, differentiation and mineralization were observed. RESULTS: The serum stimulated the cell proliferation, enhanced alkaline phosphatase activity and increased the number of mineralized nodules in the cultured osteoblasts. CONCLUSION: Kidney-tonifying traditional Chinese drugs can stimulate the proliferation, differentiation and maturation of cultured osteoblasts in vitro.