Effects & Mechanism Of Nourish-yin-and-clear-luo Treatment Rule On Bone Destruction Induced By Rheumatoid Arthritis

1 Background and objectiveBone destruction is an important pathological process of rheumatoid arthritis (RA) that no special therapeutic drugs to treat with and treatment of traditional Chinese medicine (TCM) demonstrates favorable as well as potential effects to it. Yin-deficiency-and-Luo-heat is a leading theory to explain the pathogenesy with TCM and Nourish-Yin-and-Clear-Luo treatment rule as well as Qingluo-Tongbi formula (QLT) is effective to the symptoms related. The object of this study is to explore the effects and mechanism of treatment to bone destruction induced by RA with nourish-Yin-and-clear-Luo rule as well as QLT.2 Items and methodsLook up to the books reserved by the library of Nanjing University of Chinese Medicine and retrieve the digital documents reserved by CNKI through Internet as for Chinese culture; meanwhile, as for English culture, digital documents reserved by Highwire Press of Stanford University and Pubmed are retrieved through Internet, furthermore, some documents are gotten from China Academic Library & Information System. In order to study TCM theory further, following the Supervisor and famous TCM doctors in clinical practices are performed.Three dose levels including small, medium and large of QLT are set up and ripterygium wilfordii polyglycosidium (TⅡ) is used as the positive control, in addition to model group and normal control, there are 6 groups involved in the experiments. II-type collagen induced arthritis (CIA) rats model are applied and tissue slides of their joints are observed under microscope to detect the effects of drugs on histological changes. Effects of drugs on TNF-αlevel secreted by peritoneal macrophage (MΦ) of rat models are tested by ELISA and IL-1 level by hymocyte proliferation method as well as MTT. Patients who are in the active stage of RA and have Yin-deficincy-and-Luo-heat symptoms are selected and their peripherial bloods are extracted as well as treated with serum containing drugs, then expressions of RANKL are tested with flow cytometry as per direction. Joint synovial membrane tissues are obtained from the patients who subjected to joint replace practices and fiber-forming synovial cells (FLS) are separated and cultured, then serum containing drugs to treat them and examine the FLS proliferation with MTT and expression of RANKL mRNA with RT-PCR respectively. Myelomonocytes from limb bones of SD rats are separated and osteoclast-like cells (OLC) are produced through stimulation with M-CSF as well as RANKL. OLCs are identified with TRAP staining, scrum containing drugs are added to treat with, then activity of OLCs is examined by MTT and TRAP reagent kit, moreover, ability of cells to make point pits on bone are examined to detect the effects of drugs. 3 ResultsAdvances in treatment research of bone destruction induced by RA were reviewed and key roles of inflammatory factors including IL-1, TNF-αand RANKL/RANK system as well as osteclasts playing in bone destruction had been surveyed. The importance of pathogenesis of Yin-deficiency-and-Luo-heat to RA patients who could be diagnosed as heat-type had been described, meanwhile, the viewpoints that the treatment rule should be applied as nourish-Yin-and-clear-Luo as well as QLT is an effective formula to this symptoms had been discussed.The joint synovium of rats in normal control group had no symptoms related to inflammation or evidence of bone destruction observed, however obvious inflammatory symptoms were observed in the joints, and articular cartilage erosion as well as focal necrosis was observed in the model group. The inflammatory symptoms of rats treated with TⅡor QLT were inhibited obviously. Comparing with the normal control group, IL-1 secreted by pMΦof model group was higher significantly, and TⅡas well as QLT suppressed the secretion. T lymphocytes within peripherial blood extracted from patients in active stage of RA after stimulation of PHA and being cultured, the RANKL expression of the normal control was 14.933±7.00%, under the treatment with serum containing drugs, expressions of RANKL of each groups were TⅡ:6.41±2.56%, QLT small dose:11.62±4.28%, medium dose: 8.79±4.08%, large dose:6.47±3.16%, respectively. Meanwhile, serum containing drugs demonstrated function to inhibit RANKL expression of synovium fiber-forming cells similarly. Myelomonocytes obtained from rat limb bone cavities after being induced by M-CSF as well as RANKL were found to grow as OLC. And serum containing drugs including TⅡor QLT were able to put down the activity and the energy of OLC to make point pits in the surface of bones.4 ConclusionNourish-Yin-and-Clear-Luo treatment rule as well as QLT demonstrates effects to inhibit the inflammatory reaction of RA, to suppress the expression of cell factors related to bone destruction such as IL-1, TNF-αand RANKL, to depress the activity of osteoclasts and block the development of bone destruction induced by RA. The mechanism that the treatment rule as well as QLT to cure the bone destruction induced by RA may be defined as the inhibition to deregulation of RANKL expression, blockade to the combination between RANKL and RANK and depression to the activity as well as proliferation of osteoclats.