Therapeutic Mechanism of Huangqi Guizhi Wuwutang on Rheumatoid Arthritis / 中国实验方剂学杂志

ObjectiveTo reveal the mechanism of action of Huangqi Guizhi Wuwutang in the treatment of rheumatoid arthritis by pharmacological research based on its clinical application. MethodThe collagen-induced arthritis （CIA） rat model was established by injecting bovine type Ⅱ collagen and Freund's adjuvant at the tail， and was treated with different concentrations of Huangqi Guizhi Wuwutang. The rats were randomly divided into blank group， model group， methotrexate （0.9 mg·kg-1） group， and Huangqi Guizhi Wuwutang low- and high-dose （5.13， 20.52 g·kg-1·d-1） groups， with continuous intragastric administration for 4 weeks. The degree of joint swelling， weight， degree of foot swelling and arthritis index score were determined and the pathological changes of ankle joints were detected by hematoxylin and eosin （HE） staining to observe the therapeutic effect of Huangqi Guizhi Wuwutang on rheumatoid arthritis. In addition， enzyme-linked immunosorbent assay （ELISA） and Western blot were used to measure the expression of interleukin 1β （IL-1β）， interleukin 6 （IL-6）， interleukin 10 （IL-10） and tumor necrosis factor-α （TNF-α） in serum and the expression of nuclear factor kappa-B （NF-κB） pathway related proteins in synovial tissue， respectively to clarify the molecular mechanism of Huangqi Guizhi Wuwutang in the treatment of rheumatoid arthritis. ResultCompared with the conditions in blank group， the body weight and IL-10 level were decreased （P<0.01）， and the degree of foot swelling and arthritis index score， the levels of IL-1β， IL-6 and TNF-α， and the expression of NF-κB pathway related proteins were increased （P<0.01，） in the model group， with impaired morphology and function of the ankle joint. Additionally， compared with the model group， Huangqi Guizhi Wuwutang low- and high-dose groups had increased body weight of rats and IL-10 level （P<0.01）， and reduced degree of foot swelling and arthritis index score （P<0.05， P<0.01）， levels of IL-1β， IL-6 and TNF-α （P<0.01） and expression of NF-κB pathway related proteins （P<0.05， P<0.01）， with improved function and morphology of the ankle joint. ConclusionHuangqi Guizhi Wuwutang can significantly alleviate joint inflammatory injury by down-regulating NF-κB pathway and reducing the inflammatory response in CIA rats.

Effect of Sanbi Granules on TLR4/MAPKs/NF-κB Signal Pathway in Type Ⅱ Collagen Induced Arthritis Rats / 中国药学杂志

OBJECTIVE: To discuss the effect of Sanbi granules on type Ⅱ collagen induced arthritis (CIA) rats by regulating the TLR4/MAPKs/NF-κB signal pathway. METHODS: Sixty of Wistar rats were randomly divided into normal control group (CTL group, n=10), model group (n=10), positive control group (n=10) and low dose of Sanbi group (n=10), middle dose of Sanbi group (n=10), high dose of Sanbi group (n=10). The collagen induced arthritis (CIA) model of rats was adopted and treated for 20 days by intragastric administration from 2 weeks after primary immune. After exposure to sanbi for 35 d, the rats status, paw swelling, arthritis index (AI) and pathological change of synovial tissue were observed. The serum IL-1β, IL-6, tumor necrosis factor α (TNF-α) levels were detected by ELISA. And the expressions of Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB) (p65), p-NF-κB (p65), p38, p-p38, extracellular signal-regulated kinase 1/2 (ERK1/2), p-ERK1/2, c-Jun NH2-terminal kinase (JNK), p-JNK mRNA or proteins in synovial tissues were detected by qRT-PCR and Western blot. RESULTS: At the end of experiment, compared with model group, the paw swelling degree and arthritis index (AI) of CIA rats in DXM group and low, middle, high dose of Sanbi groups were lower (P0.05). Besides, compared with CTL group, TLR4, p-NF-κB (p65), p-p38, p-ERK1/2, p-JNK mRNA and proteins in synovial tissues of CIA rats in model group, DXM group and low, middle, high dose of Sanbi groups were higher (P<0.05). And these mRNAs and proteins in DXM group and low, middle, high dose of Sanbi groups were lower than these in model group, particularly in DXM group and high dose of Sanbi group (P<0.05). CONCLUSION: There are significant evidences that Sanbi granules could protect joint synovial tissues injury by down-regulation TLR4/MAPKs/NF-κB signal pathway on CIA rats.

Comparative study on dose-toxicity-effect of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets on CIA model rats / 中国中药杂志

The aim of this paper was to compare the properties of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets from dose-effect-toxicity on type Ⅱ collagen-induced arthritis( CIA) in rats. SD rats were randomly divided into eight groups,including normal group,model group,Tripterygium Glycosides Tablets groups( 1 times equivalent dose 0.009 g·kg-1,4 times equivalent dose 0.036 g·kg-1,16 times equivalent dose 0.144 g·kg-1),Tripterygium wilfordii Tablets groups( 1 times equivalent dose 0.007 5 mg·kg-1,4 times equivalent dose 0.030 mg·kg-1,16 times equivalent dose 0.120 mg·kg-1). Beginning on the first immunization,Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets administered intraperitoneally once a day. After the second immunization,the symptoms such as redness and swelling of joints were observed,and the clinical score and incidence of arthritis were evaluated. HE and Masson staining were used to examine the histopathological changes of joints. The expression level of anti-type Ⅱ collagen antibody Ig G in serum was detected by ELISA,routine testing of blood components,the concentration of ALP( alkaline phosphatase),ALT( alanine aminotransferase),AST( aspartate aminotransferase),GGT( gamma-glutamyltransferase),TBi L( total bilirubin),CRE( creatinine) and UREA( urea) in serum were detected by enzymatic assay. The rate of sperm deformity in the epididymis was evaluated under light microscope. The extent of damage to the testis and ovarian tissue was assessed by HE staining. Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets attenuated the inflammation,redness,swelling and deformity of joints and reduced the clinical score and incidence of arthritis in CIA rats. Meanwhile,it also exhibited obvious reduction in all pathological features such as joint synovitis,pannus,cartilage erosion and bone destruction. Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets reduced Ig G in a dose-dependent manner,and Tripterygium Glycosides Tablets is better than Tripterygium wilfordii Tablets( P<0.05 or P<0.01). The high doses of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets could significantly increase the organ coefficient of liver and spleen and reduced RBC and HGB in CIA rats( P<0.01),and severity leading to death. Gastric mucosal injury and morphological changes of liver and kidney were not observed in CIA rats of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets treatment group. The 4 and 16 times doses of Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets could significantly increase serum ALT,GGT and decrease CRE( P<0.05 or P<0.01). Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets could increase the sperm deformity rate and damage the testicular seminiferous tubules of CIA male rats. Severity increased with dose and time increasing. The effect of Tripterygium Glycosides Tablets( 16 times) is more significant than Tripterygium wilfordii Tablets( 16 times). Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets significantly delayed onset of arthritis and inhibited the paw edema and arthritic score. Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets also caused male reproductive damage,high dose affected hematopoiesis,and maximum dose leading to death. Tripterygium Glycosides Tablets and Tripterygium wilfordii Tablets all depended on dose-effect-toxicity manner. Anti-arthritis effect of Tripterygium Glycosides Tablets is better than Tripterygium wilfordii Tablets,but the toxicity of Tripterygium Glycosides Tablets maximum dose is more obvious. The relevant conclusions of our study will provide experimental references for clinical rational use of drugs,and further clinical studies are needed to confirm our conclusions.

Study on inhibitory effects of Triperygium Wilfordii Polyglucoside on Dipeptidyl peptidase I and regulatory mechanism / 中国免疫学杂志

Objective:Dipeptidyl peptidase I(DPPI),a lysosomal cysteine protease for serine proteases activation,highly expressed in granule immune cells.This study used collagen induced arthritis(CIA) rat model to investigate the effects of Triperygium Wilfordii Polyglucoside(TWP) on DPPI activity and the pharmacological mechanism in RA treatment.Methods:Rats were divided into four groups randomly,the blank control group,the CIA model group,the high dose (5.0 mg/100 g body-weight) and low dose(2.5 mg/100 g body-weight)treatment group.Bovine collagen-Ⅱ plus complete Freund′s adjuvant injected twice in rats.Physical assessments were carried out.12 days post-injections,the rats of treatment group were intragastric administered with TWP every day.The rats were killed after two week administrations.Serum and synovial membrane homogenates were collected and DPPI activity was detected by fluorescence substrate.Joint HE staining and cell counting were carried out,Zymography was used to detect the MMP-2/9 activity in synovial fluids.Total protein in synovial membrane homogenates were measured by BCA method.Results:TWP could reduce the number of CIA synovial tissue mast cells,inhibited DPPI activity in the synovial fluids and in serum.The expression levels of MMP-2/9 activity and synovium total protein content were also reduced by TWP.Conclusion:Triperygium Wilfordii Polyglucoside has inhibitory effects on DPPI activity on CIA rats,which might be the one of the pharmacological mechanisms in RA treatment.