By : Huifang Zhao, Ph.D.

Rheumatoid arthritis (RA) is a chronic inflammatory disease, which characterized with proliferation of synovial membrane and progressive destruction of articular cartilage. It was reported that the extensive proteolytic degradation of articular cartilage is commonly associated with invasive hypertrophied synovium, which express a large quantities of proinflammatory cytokines including interleukin-1 beta (IL-1), IL-2, IL-4, IL-6, IL-8, TNF alpha, etc. Among these cytokines IL-1 and TNF play a pivotal role in the progression of inflammation and the degradation of joint connective tissues. They induce the proliferation of synoviocytes and enhance their production of prostaglandins (PGE2), metalloproteinases (MMPs), and cytokines; stimulate ascular endothelial cells to express adhesive molecules; increase production of acute phase reactants such as fibrinogen and c-reaction protein (CRP); effluence osteoclast /osteoblast function, those leads to the constantly inflammation of joints, invasive pannus formation, cartilage damage and joint destruction.

From the view point of Chinese Medicine, the etiology and pathology of RA is due to the weakness of Qi and Blood, the deficiency of Liver and Kidney essence and the invasion of wind-cold-damp. The prolonged illness generates the pathogenic heat in the joints, blockages the circulation of qi and blood, leads to a very complicated situation-the combination of cold and heat, the wind and damp, the deficiency and excess, the imbalance of Yin and Yang. TBL, Chinese Medicine, composed of Nan She Teng (Celastrus orbiculatus Thunb.). Ji Xue Teng *Millettia dielsianaa Harmsex Diels), has the function of dispelling wind-damp, clearing heat, detoxify, unblocking the channels, invigoration blood and nourishing blood, showed its antiinflammatory, immune-modulating function, relieving pain and inhibiting the destruction of joint bones in the clinical and experimental examination.

CLINICAL STUDY

137 cases of RA were diagnosed according to the RA diagnostic criterion modified by American Rheumatoid Arthritis Association (ARA) in 1987, were obtained from the Internal Medicine II Dept. and the specialist’s clinic, 1st Affiliated Hospital of Guaangzhou University. Patients were of TBL tablet, 0.25g/tablet, 4 tablets, three time a day; Control group accepted the treatment of evaluation, ESR, rheumatoid factor (RF) titer, CRP, IgG, IgM. peripheral blood T-lymphocyte subpopulation distribution. PQMS medical statical software was employed to analyze T hypothesis of the experimental date.

The results showed that TBL had a remarkable effect on improving clinical symptoms, stopping pain, eliminating swelling and morning rigidity, reducing ESR, CRP, RF titer, etc. Also showed the decreasing the ratio of CD4/CD8 and the number of B cell, regulating the balance of Th/Ts. and suppressing the hyperactivity of humeral immune. There is significant difference comparing with MTX group. These findings suggest that the mechanism of the TBL on treatment of RA has a close relationship with its effect of immunoregulation, blockage the immune pathology of RA to improve the illness.

LABORATORY STUDY

In this study, type-II collagen induced arthritis (CIA) and adjvant arthritis (AA) in DBA/1J mice, Lewis rats animal models were used, Indomethacin (IM) and Methotrexate (MTX) serve as positive controls. TBL was suspended in distilled water and orally administered from 4 weeks after the first immunization. The dose of each drug are TBL 2g/kg. d-1, IM 2mg/ky. d-1, (MTX) 2mg/kg.w-1. Saline was given to the negative control and C II-immunized mice during the same period.

The following studies were employed to prove the hypothesis of immune-regulatory of TBL.

1. Symptomatic score evaluation of CIA mice

2. Bone destruction evaluation of CIA mice

3. Anti-IgG antibodies in serum of CIA mice (ELISA)

4. The ratio of CD4, CD8 positive cell and t-lymphocyte in spleen of CIA mice (Immunohistochemistry)

5. The expression of Metalloproteinase MMPs in synovium tissues of CIA mice (Immunohistochemistry)

6. The activity of IL-1, TNF alpha and PGE2 in synoviocytes culture supernatant (ELISA)

7. The expression of IL-1 beta messenger RNA of synovial cell in AA rats (RT-PCR)

8. Mast cell and mast cell chymase activity and their protein level assay (Enzyme activity)

9. Mice mast cell chymase-2, -4 and -5 (MMCP-2, -4 and -5) mRNA expression in arthritic paw skin of CIA model (RT-PCR)

Studies clarified the molecular immunologic pharmacology mechanism of TBL on the antirheumatic disease on the cell, molecular and gene levers respectively. The results showed that: TBL inhibits the synovial cellular proliferation, fibrous proliferation, inflammatory cell infiltration in knee joints of CIA mice and rats, significantly improve the symptomatic score and bone destruction; decresases the titer of anti-CII antibody in serum; decreases and inhibits the production and activity of PGE2 and MMPs in culture supernatant and synovial tissue; inhibits the bioactivity and gene expression of proinflammation cytokines IL-1 beta, TNF alpha in synovial tissue; immune-modulates the interrelation of T, B lymphocytes, Mast cells in arthritic joints; reveals the role of mast cell on pathogenesis of arthritis rheumatoid model, and TBL inhibits the mast cell chymase activity, MMCVP-2, -4 and -5 mRNA expression in arthritic paw skin of CIA model. The novel information on pathogenesis of RA and mechanism of TBL may contribute to the further developing novel drugs RA with less side-effects in the near future.

REFERENCES

1. The Clinical Research of Rheumatoid Arthritis Treated with TBL, New Journal of TCM 1997; 29 (9): 13-21

2. Suppressive Effects of a herb formula, TBL, on Type II Collagen induced arthritis in DBA/1J mice. Clinical and Experimental Pharmacology and Physiology 23,650-662, 1996.

3. Effect of TBL on the free radical in patients with rheumatoid arthritis, Journal of Guangzhou University of TCM 14:222-230, 1997

4. The clinical and experimental study of rheumatoid arthritis treated with TBL, The Practical Journal of Integrated Traditional and Western medicine 18:34-41. 1998

5. Effect of TBL on IL-1 beta, TNF alpha mRNA expression in synoviocytes in Adjuvant-induced arthritis rat, Cvhinese Journal of the Practical Chinese With Modern medicine 12:628-630, 1999

6. Effect of TBL on the influence of proliferous reaction of synovioblast of rats, New Journal of TCM 31:37-40,1999

7. Primary culture of synoviocytes of rats with adjuvant arthritis and its secretary function, Journal of Guangzhou University of TCM 116:60-66. 1999

8. Effect of TBL on synoviocyte function in adjuvant arthritis rats, Chinese Journal of Integrated Traditional and Western medicine 19: 167-170, 1999

9. Role of mast cells on pathogenesis of arthritis rheumatoid model, Biomedical Gerontology 22:1-2, 1998

10. Role of mast cells on pathogenesis of arthritis rheumatioid model, presented at the 21st Biomedical Gerontology Conference held in Jun. 17-19, 1998, Tokyo, Japan.

11. Potential pathogenic role of mast cells in type II collagen-induced arthritis of rat, presented at the 72nd Conference of Pharmacology held in Mar. 22, 1999, Saporo, Japan.

12. Effects of orally applied crommolyn prodrug on the II collagen-induced arthritis in DBA/1J mice, presented at the 4th International Conference of Inflammation held in June 27-30, 1999, Paris.

* The study was supported by the following foundation:

1. Assessment of Chinese Medicine for incurable Diseases-Japan-China Collaborative Clinical and pharmacological Assessment, supported by Japanese government international cooperation research foundation.

2. Immune-mechanism of Chinese medicine in treating rheumatic diseases, supported by the vital project of China National Foundation of Nature Science. (The highest national level and multiple joint research project on China)

3. Effect of TBL on the expression of synovium IL-1 beta, TNFaalpha mRNA on CIA mice, supported by China National Foundation of Nature Science.