ABSTRACT
BACKGROUND: Chinese medicine is rich in recognition and treatment for rheumatoid arthritis(RA) and has achieved attractable progression in researches of recent several years,characterized by variable therapeutic methods, definite therapeutic effects and few toxic side effects. Therefore, it is important scientifically and socially to further search for the effective drugs and methods for the treatment of RA in the field of Chinese medicine. There are few reports on whether Chinese herbs inhibit or block the destruction of cartilage and bone and promote their metabolism and repair for damage or not by variable approaches,targets and links in RA occurrence.OBJECTIVE: To probe into the mechanism of Chinese herbal tongbiling on cartilage destruction.DESIGN: Complete random design and control experimental study based on the experimental animals.SETTING: Golden Chamber teaching and research room of guangzhou university of traditional Chinese medicine.MATERIALS: The experiment was performed in Comprehensive Experimental Room of First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine from August 2002 to December 2003,in which,35female Wistar rats were employed,clean grade,weighted(110±20) g,provided by Animal Experimental Room of First Military Medical University of Chinese PLA, with qualified animal No. 2002A041.After 3 days adaptive breeding,Wistar rats were randomized into normal group,model group and total alkaloidal of tongbiling group(TBL group). TBL was extracted in Phytochemistry Room of Tropical Disease Institute of Guangzhou University of Traditional Chinese Medicine.METHODS: The animal model was prepared on rats with collagen-induced arthritis. After treated with gastric infusion in groups,the pathological sections were done on phalangeal joint of rat and chondropathy was observed after routine HE staining. The chondrocytes were cultured. The chondrocytes were stimulated with rrIL-1β in vitro and resulted in matrix decomposition of chondrycyte to prepare pharmaceutical model,TBL of various concentrations added and dexamethasone(DXM) was applied in the control. After 48 hours culture,the upper clear solution if cell was collected and glycosaminoglycan (GAG) content was measured with alcian blue method,and nitric oxygen (NO) content was assayed with method of aqua fortis reduction.MAIN OUTCOME MEASURES:①Primary results: Effect of TBL on phalangeal chondropathy in rats with collagen-induced arthritis and on matrix decomposition of chondrocyte induced by rrIL-1β.②Secondary results: Effect of TBL on induction of NO synthesis from chondrocytes by rrIL-1β.RESULTS:①It was indicated in evaluation on HE pathological section of phalangeal joint: The severity of cartilage destruction in TBL group was less than that in model group(P<0.05).②TBL of various concentrations and DXM inhibited remarkably the decomposition of choncrocyte proteoglycan (P<0.01) and NO synthesis (P<0.01).CONCLUSION: TBL inhibits phalangeal cartilage destruction of rats with collagen-induced arthritis. It is indicated in the results of experiment in vitro that TBL is against the decomposition of choncrocyte proteoglycan,which is probably achieved by inhibiting NO synthesis.
ABSTRACT
[Objective] To investigate the effect of Erteng Mixture (EM) on activated T-cell CD69 expression and Jurkat cell cycle in rheumatoid arthritis (RA) and to explore its possible cellular immunoregulation mechanism. [Methods] Jurkat cells were cultured with different kinds of components extracted from EM. After 72 hours, proliferation rate of Jurkat cell were detected with thiazolyl blue colorimetry (MTT) to screen the active component. Whole blood from RA patients were incubated in RPMI-1640 culture with various concentrations of chlorofonn component extracted from EM or with Genistein. Thirty minutes later, phytahematoagglutinin (PHA) were added successively into the culture. After 24 hours, CD69 expression rate of T lymphocytes activated by PHA was analyzed by flow cytometry. Jurkat cells were cultured with various concentrations of chloroform component extracted from EM and with methotrexate ( MIX) for 48 hours. After then, Jurkat cell cycle was analyzed by flow cytometry. [ Results ] Chloroform component extracted from EM had the strongest inhibitive effect on proliferation of Jurkat cells. Chloroform component (10 mg/L and 20 mg/L) could markedly inhibit CD69 expression on PHA-activated T lymphocytes. Chloroform component 10 mg/L blocked Jurkat cell cycle at G1 phase, which differed from MTX blocking Jurkat cell cycle at S phase. [Conclusion] Chloroform component extracted from EM could significantly inhibit the early activation of CD4 T lymphocytes in RA and inhibit the energy and material synthesis in T-cell, thus hindering DNA synthesis and decreasing the cellular G2 phase transformation. This study will provide an experimental basis for clinical application of EM in treating RA.