Clinical Effect Of Meridian-Sinew Release Therapy On Knee Osteoarthritis And Its Effect On TLR4 Signaling Transduction Pathway

Purpose1.By use of drugs, drugs combine electroacupuncture, drugs combine Meridian-Sinew Release for treatment of the knee osteoarthritis, to compare these 3 different treat methods, to evaluate the clinical efficacy of the Meridian-Sinew Release therapy.2. Experimental Study of TLR4/MyD88/NF-κB signaling transduction pathway system on the rabbit knee osteoarthritis model induced by the papaya protease, clarify the mechanism of Meridian-Sinew Release therapy for Knee Osteoarthritis treatment.Method1. Clinical Research:According to the diagnostic criteria, inclusion criteria and exclusion criteria, choose 60 outpatients and inpatients in traditional Chinese medicine department, guangzhou general hospital, who had been diagnosed as knee osteoarthritis. The research objects were divided into three groups randomly:drugs group, drugs combine electroacupuncture group, drugs combine Meridian-Sinew Release therapy group. Abserve the clinical and lab indexes of the three groups before the cure and 1 month after treatment, and evaluate clinical curative effects according to the WOMAC rating scale. Finally, the indicators of the 3 groups were compared between groups, statistical data analysis, draw the conclusion.2. Experimental Research:Select 24 6 months old healthy clean New Zealand rabbits, per weight 2kg±0.5kg, half male and half female, choose 6 rabbits as blank control group, use 1.6%papain liquor inject in the left knee joint cavity of the other 18 rabbits on Day 1, Day 4 and Day 7, to make the Knee Osteoarthritis model, and the blank control group use 0.9% normal saline injected in the left knee joint;6 weeks after injection, divide the 18 rabbits into 3 groups randomly:Meridian-Sinew Release therapy group, electroacupuncture group, model group, (n=6). Raise each rabbit in single cage, the size of the cage can ensure its activities freely. Temperature of the lab keeps on 23℃-26℃ constantly, humidity on 60%, ultraviolet disinfection regularly, dieting freely. In the process of making models, if appeared such as foot ulceration, bitten foot by themselves, fracture, skin infections,diarrhea, etc., would be removed. Choose 1 rabbit from each group randomly to take the X-ray of left knee, and compare with the blank control group, other 3 group have the changes, like joint space narrowing, osteophyte formation, sclerosis of subchondral bone, under the cartilage capsule be changed;And according to the behavioral indexes, suggest that the models were made successfully.1 week after finished making the models, begin the treatment for electroacupuncture group and Meridian-Sinew Release therapy group for 4 weeks. The electroacupuncture group treatments are on alternate days 1,3 times a week;The Meridian-Sinew Release therapy once a week which lasting for 4 weeks. After the treatment finished for a week,use air embolism method to kill all the rabbits. Observe the articular cartilage and synovial pathological manifestations in each left knee, and the pathological performance of the rabbits’cartilage pieces by light microscopy. Testing the gene and protein expression of TLR4, MyD88, NF-κ B by PCR and Western Blot. Finally, data analysis performed with statistical process.Results1.Clinical Research:Acccording to the statistical analysis of VAS scores and WOMAC scores, the results show that before and after the treatment, the differences of drugs group, drugs combine electroacupuncture group, medicine combine Meridian-Sinew Release therapy group have statistical significance(P＜0.05);Compare with the different groups, the curative effect of drugs combine Meridian-Sinew Release therapy group is better than the drugs group or drugs combine electroacupuncture group(P＜0.05), the differences have statistical significance.2. Animal experiments:①Give treatment for 4 weeks after make the model, generally observe in rabbit articular cartilage and synovial membraneafter after finish the treatment,there’s no joint swelling in blank group, and synovial is pink, no hyperemia, edema, smooth articular surface, no neoplasm, cartilage color is bright, clear liquid, no joint turbidity, capsular thickening;In model group, left knee joint swelling, synovial thickening, open hyperemia, articular cartilage, pale dull, uneven surface of bone is visible in a number of coarse, osteophyte formation, joint fluid yellow muddy, thick, joint capsule thickening;In electroacupuncture group, left knee joint swelling. After opening with a few slight thickening of synovial hyperemia, cartilage pale color mild bone defect, slight osteophyte, joint fluid is clear, mild thickening of articular capsule;In Meridian-Sinew Release therapy group, left knee tendons swelling mildly, synovial proliferation compared with the blank group, electroacupuncture group light, no marked hyperemia, pale color of cartilage, mild bone defect, visible a few osteophyte, synovial fluid than the blank group, joint capsule thickening mildly. ②Light microscope observation of rabbits cartilage HE staining pathological results:Meridian-Sinew Release therapy can significantly inhibit the cartilage cells be destructed, and promote the proliferation of cartilage cells. ③Compare with the blank group, TLR4, MyD88, NF-κ B gene and protein expression significantly increased in model group;Compare with electroacupuncture group and model group, the gene and protein expression of TLR4/MyD88/NF-κB decreased in Meridian-Sinew Release therapy group, the difference have statistical significance (P＜0.05).Gonclusion1. Clinical Research:By observing and comparing drugs combine Meridian-Sinew Release therapy, drugs combine electroacupuncture, or the drugs to treat the Knee Osteoarthritis, we can conclude that each method can release the pian and swell, improve the activity function of joint. Compared with the electroacupuncture and drugs treatment group, Meridian-Sinew Release therapy can relieve the symptom more quickly and have prominent therapeutic effect, which is worthy to promote in clinical.2.Animal experiments:By observing the contrast between each group of rabbit knee joint behavior index, articular cartilage and synovial pathological manifestations, HE staining of cartilage pieces observed by light microscope, testing the gene and protein expression of TLR4, MyD88, NF-κB by PCR and Western Blot, we infer that the Meridian-Sinew Release therapy may regulate TLR4/MyD88/NF-κ Bsignal transduction pathway, inhibit the inflammatory reaction such as synovial proliferation and joint swelling or pain, relieve the symptoms like dysfunction of joint activities and stiffness in the morning, to get the purpose of control the further deteriorate of the disease.