ABSTRACT
Objective To observe the effects of the drug pair of Rhizoma Polygoni Cuspidati ( Huzhang) and Ramulus Cinnamomi ( Guizhi) on the Toll-like receptor 4 mediated myeloid differentiation factor 88 ( TLRs/MyD88) signaling pathway of rats with acute gouty arthritis induced by monosodium sodium urate (MSU) , so as to explore its therapeutic mechanism. Methods Forty-eight male SD rats were divided into normal group, modele group, blank plasmid group, positive plasmid group, Huzhang- Guizhi herb-pair (7 g/kg) group, and Huzhang-Guizhi herb-pair ( 7 g/kg) siRNA group, 8 rats in each group. The normal group, plasmid groups and model group were given physiological saline, and the left groups were given the corresponding drug by intragastric administration for 10 continuous days ( once daily ) . On the seventh day of intragastric gavage, acute gouty arthritis were induced by injection of MSU into the rat ankle joint, and normal group was injected with the samevolume of normal saline. Positive plasmid group and Huzhang-Guizhi herb-pair siRNA group were injected with the constructed siRNA-TLR4 plasmid targeting TLR4 gene ( TLR4-siRNA) to inhibit the in-vivo TLR4 gene expression. Pathological changes of the synovial tissues were detected, the contents of peripheral blood tumor necrosis factor alpha ( TNF-α) and interleukin 1 beta ( IL-1β) were detected by double antibody sandwich method, and the mRNA and protein expression levels of TLR4, MyD88, TNF receptor-associated factor 6 ( TRAF-6) in peripheral blood mononuclear cells of rats were detected by real-time fluorescence quantitative polymerase chain reaction ( PCR) and Western blot methods. The nuclear factor kappa B ( NF-κB) p65 immunoactivity was assayed by immunohistochemistry. Results Compared with the normal group, the model group had obvious hyperplasia of synovial cells and the inflammatory cell infiltration ( dominated by lymphcytes and monocytes) , and had amount of cellulose adhesive on the synovial membrane surface. Compared to the model group, positive plasmid group, Huzhang- Guizhi herb-pair group and Huzhang-Guizhi herb-pair siRNA group could obviously relieve the inflammatory cell infiltration, and improve synovial cell proliferation reaction. Compared to the normal group, serum levels of TNF-α and IL-1β, and the expression levels of TLR4, MyD88, TRAF-6 mRNA and protein in the peripheral blood mononuclear cells as well as the synovial NF-κB p65 ex pression in the model group were significantly increased ( P<0.01). Compared to the model group, positive plasmid group, Huzhang-Guizhi herb-pair group and Huzhang- Guizhi herb-pair siRNA group showed significant decrease in the levels of TNF-α, IL-1β, TLR4 MyD88, TRAF-6 and NF-κB p65 ( P<0.05 or P<0.01) . Conclusion Huzhang-Guizhi herb-pair can regulate the cytokines of the synovial membrane tissue in acute gouty arthritis rats, which may be related with its effect on inhibiting abnormal activation of TLR4-MyD88-NF-κB pathway in synovial tissue.
ABSTRACT
Objective To observe the clinical efficacy of Qingre Liangxue Recipe (QLR), a herbal prescription with the actions of clearing heat and cooling blood, for the treatment of gouty arthritis. Methods Sixty-five cases of gouty arthritis patients were randomly divided into treatment group (N=32) and control group (N=33). Both groups were given oral use of colchicine during the acute stage and being suspended when the pain was relieved, and then were given oral use of allopurinol tablets during the remission stage. Additionally, the treatment group was given oral use of QLR all through the acute stage and remission stage. The treatment lasted for 4 weeks in both groups. After treatment, clinical efficacy was evaluated in the two groups. The number of acute relapse cases and the changes of blood uric acid level were observed. Results At the end of the first treatment week, the treatment group had a total effective rate of 96.9%, significantly higher than that (81.8%) of the control group. At the end of the fourth treatment week, the treatment group had a total effective rate of 100.0%, significantly higher than that (90.9%) of the control group. The treatment group had better therapeutic effect than the control group at the end of the first or the fourth treatment week, the difference being significant ( P<0.01). The number of acute relapse cases and the serum uric acid level were lower in the treatment group than those in the control group during 4 treatment weeks or 12 weeks after treatment suspension, the difference being significant ( P<0.01). Conclusion QLR combined with anti-gout medicines can effectively relieve symptoms of gouty arthritis, reduce incidence of acute relapse, and have long-term effect on lowering blood uric acid level.
ABSTRACT
Objective To investigate the influence of serum containing Qingre Chubi Decoction ( QCD) on the THP-1 cell viability and the release of interleukin 1 beta ( IL-1β) stimulated by monosodium urate crystals in vitro. Methods The cultured human monocyte THP-1 strain were divided into blank serum group, model control group, and high-, middle- and low-concentration ( volume fraction being 20%, 10%, 5%) QCD-containing serum groups. Except for the blank serum group , the other groups were all given 500 mg/L of monosodium urate crystals. On culturing hour 0, 12, 24 and 48, THP-1 cell viability was tested by methy1 thiazolyl tetrazolium celorimetry ( MTS) method. On culturing hour 48, the content of IL-1β in the supernatant of THP-1 cells was detected by enzyme-linked immunosorbent assay ( ELISA) . Results The THP-1 cell viability in various groups was increased along with the prolongation of culturing time. The THP-1 cell viability in the model control group was increased as compared with that in the blank serum group at different time points (P<0.05 or P<0.01) . And the content of IL-1β in the model control group was increased significantly as compared with that in the blank serum group on culturing hour 48 (P<0.01) . The THP-1 cell viability in various QCD-containing serum groups on culturing hour 12 and 24, and in high- and middle-concentration QCD-containing serum group on culturing hour 48 was decreased significantly as compared with that of the model control group at the same time point ( P<0.05 or P<0.01) . The content of IL-1β in various serum containing QCD groups was markedly decreased as compared with that in the model control group on culturing hour 48 ( P<0.01) . Conclusion Serum containing QCD can inhibit the viability of THP-1 cells stimulated by monosodium urate crystals, and the possible mechanism is related with the inhibition of IL-1 release.