Study On The Anti-inflammatory And Anti-rheumatic Mechanism Of Panax Japonicus Saponin

Rheumatoid arthritis (RA) is a chronic, inflammatory and systemic autoimmune disease, which joints and joint's surrounding tissue non-suppurative inflammations are its main features. It becomes the second cause of loss ability to work behind the ischemic heart disease. The traditional treatment of RA drugs are mostly single pathway or target, while there are certain side effects, so development of efficient, low toxicity, the role of multi-target drug is particularly important and urgent.Panax japonicus (Panax japonicum CA Mey.),a traditional chinese medical herb, belongs to Araliaceae. It has strong tonic, dissipate blood stasis and alleviate pain, bleeding and expectorant functions in Chinese medicine record.So it used totreated weakness after diseases, tuberculosis hemoptysis, cough, sputum, and bruises. In this study, its anti-inflammatory effects and anti-rheumatic mechanisms will be carried out syetemic researched.1) The effect of TSPJ on normal inflammatory response in mice:(1) TSPJ 50 mg/kg and 100 mg/kg group could significantly reduce the paw edema, foot redness swelling and other symptoms induced by carrageenan after inflammation 0.5～4 h (P<0.05)Vs control group.(2) TSPJ could inhibit the mouse ear edema induced by croton oil, the ear's redness, swelling and other symptoms are significantly inhibited.The high dose group of TSPJ's inhibition rate is 33.33% compared with the control group. The inhibition rate is positively correlated with the dose (P<0.05).(3) TSPJ could inhibit the increased capillary permeability induced by acetic acid on mice inhibition rate of TSPJ's high dose group(100 mg/kg) is 48.6% compared with the control group. The inhibition rate is positively correlated with the dose (P<0.05).(4) TSPJ could increasethe levels of serum GSH-PX, SOD activity and reduce serum MDA content. The inhibition rate is positively correlated with the dose(5) TSPJ could reduce the inflammatory factors (IL-1, TNF-α) in mice serum, the differences are significant (P<0.05).2) The effect of TSPJ on CIA rats'histopathological and ultrastructural stractures (1) The AI scores in CIA group are higher than normal group, three dose group's AI of TSPJ were also increased compare with the normal group at the same period, but the difference were significant compare with the CIA group at the same period.(2) The model group' feet swelling begins from d12 to the end, peak time was at d24-d28. The swelling degree of TSPJ groups were also increased compare with the normal group at the same period, but the difference was significant compare with the CIA group at the same period.(3) CIA model rats'weight-gain rate slow down or stoped at the secondary inflammatory phase (after d18), TPT treatment group also shown the weight-gain rate, TSPJ groups' growth rate were normal.(4) The immune function of CIA model could lead to higher compared with model group; the administration of TPT could decreasethe higher s spleen index and thymus index in CIA rats, the difference was significant,and TSPJ group alos decline differences but no significant.(5) TSPJ can improve joint damage of CIA:thre dose groups of TSPJ can reduce the levels of congestion and edema of soft tissue joint, toe joint volume and scores; reduce synovial hyperemia, edema of the degree; significantly inhibit the CIA rats' synovial hyperplasia; inhibit the inflammatory cell' infiltration; inhibit chondrocytes proliferation.(6) ConA-induced spleen T lymphocyte proliferation in CIA group was significantly lower than the control group, and IL-2 levels were also significantly lower, TSPJ groups can restore the CIA rat spleen lymphocytes and production of IL-2, but TPT group has no effect on proliferation of lymphocytes and production of IL-2.(7) The productions of IL-1β, IL-6 and TNF-αwere significantly higher in CIA group's PMΦthan those of normal group (P<0.05), TSPJ medium and large doses groups can inhibit the production of proinflammatory cytokines IL-1β, IL-6 and TNF-αlevels in CIA's PMΦ.(8) The synovial membrane ultrastructure:at the CIA group, the synovial cell lining layers were thickened up to 5-6 layers and disarrangement, organelle deformation, or even disappear, the cytoplasm filled with large vacuoles, the uneven distribution of nuclear chromatin. Some cells due to degeneration and necrosis to atrophy, only to see a large number of cell fragments with staggered collagen fibers. TSPJ can significantly improve the CIA synovial ultrastructure of synovial lining cell layer and cell arrangement, only occasionally a small amount of lymphocytic infiltration can be seen, A-type cells to primary lysosomes, mitochondria slightly increased, B-type Cytoplasm of cells rich in rough endoplasmic reticulum and ribosome bodies, various types of cells close to normal.(9) The ICAM-1 production (the total number of positive cells, the average black level value of the total area and integral optical density) were significantly increased in CIA model group's synovial membrane during the inflammatory process, TSPJ groups can reducethe ICAM-1 expression in synovial number, size Sum, integral optical density and average black levels, the differences are significant (P<0.05).(10) synovial cell culture and cytokine researches showed:normal synovial cells' cytokines productions are very low even no detected, cell growth rate are normal; after stimulated by LPS, the speed of cell proliferation and cytokines such as IL-1, IL-6, IL-8 and TNF-αsecretions are increased, this suggest that LPS is a good stimulant of microbial antigens, can induce normal cell proliferation and differentiation. CIA synovial cells' proliferation rate and cytokines secretions were significantly higher after LPS stimulation. Tripterygium showed a strong anti-inflammatory effects; TSPJ didn't show cell-damage in the synovial cell culture experiments,and didn't stimulate cytokine secretion, only regulate the CIA synovial cells from "super- sensitivity" status to normal.From the experimental results, TSPJ has the following parameters:(1) increasing the antioxidant capacity, reduce inflammation, tissue damage induced by free radicals; (2) to relieve inflammatory mediators such as PGE and LTB in inflamed tissue; (3) significantly decreased cell factors such as IL-1B, IL-6, TNF-αinf lammated tissues; (4) reduce the ICAM-1 generation in inflammated tissues; (5) TSPJ has 'two-way adjustment' on the immune system, so at the process of treating RA it can reduce hyperthyroidism immune function to normal, but don't produce immune dysfunction as triptolide and other immunosuppressive agents do. Therefore, TSPJ is likely to be developed as an effective, non-toxic or low toxic anti-inflammatory and anti- rheumatic drug.