| Objective:By copying the knee osteoarthritis rat model, to have a staged observation of the pathological naked-eye.Brittberg grading pathological sectionMankin scoring,X-rays of knees, and ELISA result of serum TNF-α, MMP-13 of articular cartilage,to further explore the possible mechanism of Duhuojisheng decoction in preventing and treating knee osteoarthritis degeneration, and provide a theoretical basis for clinical application and further research.Methods : 60 SD rats were randomly divided into blank group 10, experimental group 50, modeling by iodoacetic acid drug induction method, after 2 weeks the modeling success, 5 rats were randomly selected for lateral radiographs of the knee, and were killed after blood collection, the right knee joint cartilage surface condition was observed with the naked eye, and produce pathology. The remaining 45 rats were randomly divided into model group, with 15 rats,western medicine group 10, in the traditional Chinese medicine middle dose group 10, Chinese medicine high dose group 10, Chinese medicine group treated by DuhuoJisheng decoction gavage, the western medicine group received tocilizumab injection treatment, 5 rats were killed in model group at fourth week and eighth week, all were killed after 8 weeks. Weekly observations of the general condition of the rats 1-2 times and records, before death, give them lateral radiographs of the knee, fasting for 12 h before blood collection, in 2% sodium pentobarbital anesthesia in abdominal aortic blood collection, preparation of medicated serum.Observing animal articular cartilage surface in body with naked eye, making pathological section, using osteoarthritis cartilage pathological changes in Mankin score standard, evaluating the articular cartilage pathological change of each group.Results:1. general situation: the blank group is more lively, hair gloss, no joint swelling, have the normal activity and diet; the model group mental burnout less dynamic, hair less shiny, hair removal significantly, eat less, local joint swelling, had licking and limp performance and with the extension of time, was continued to aggravate. In the traditional Chinese medicine middle and high dose group, western medicine group, the performance compared with the model group, have different degrees of improvement.2. the right knee X-ray examination showed: blank group joint space is uniform, articular surface is smooth, femoral articular surface of tibial platform clearoutline; knee joint space narrowed significantly in model group, the articular surface is not smooth, cartilage destruction, defect, osteophyte formation, and as the disease progress, the extent of damage gradually increase, the stage score, showed a rising trend, P<0.01, the difference was statistically significant. Compared with the 8th week model group, the group of Western medicine,Chinese medicine in middle dose group, high dose group showed different degree of repair, P<0.01, the difference was statistically significant.3.Observation of articular cartilage in vivo naked eye: blank group articular surface was smooth, the color red, model 2 weeks group the surface of articular cartilage of rats obviously loses luster, more greyish yellow, rough surface,and a small part of crack formation, 4 weeks group joint surface irregularities, cartilage defects as the middle,synovial joint capsule, with different degree thickening, 8 week group joint surface defect, even ulcer surface,cartilage defects as deep, with joint effusion formation.According to Brittberg grading standard score, score rises gradually, the comparison among groups was significantly different, P<0.01, the difference was statistically significant. Compared with the traditional Chinese medicine, in high dose group, western medicine group than in model group 8 weeks, had different degree ofrecovery, scores were lower, the curative effect is obvious, the difference was statistically significant(P<0.01), but the high dose of Chinese medicine group and Western medicine group compared the curative effect had no obvious difference, the difference was statistically significant(P>0.05).4.Pathology and Mankin slices were under light microscope results showed that showed a score of knee osteoarthritis pathological change early and middle. Compared with second weeks, fourth weeks, 8 score increased significantly, cartilage degeneration further aggravating, the inter group comparison, P<0.01, the difference was statistically significant. Eighth weeks, compared to traditional Chinese medicine, high dose group, western medicine group and model group, scores were significantly reduced, the curative effect is obvious, the difference was statistically significant(P<0.01), high dose of Chinese medicine group with Chinese medicine dose group compared, rating decreased,the difference was statistically significant(P<0.01), but the high dose of Chinese medicine group and Western medicine compared the curative effect, no significant differences, nosignificant difference(P>0.05).5. Between the groups of serum TNF-α, MMP-13 index: with the prolonging of molding cycle,compared with the blank group, the content of model 2, 4, 8 weeks in serumTNF-α,MMP-13 assumes the trend of escalation, inter group comparison, P<0.01, the difference was statistically significant. After drug treatment, different doses of Duhuojisheng Decoction group and Western medicine group, the content of TNF-α in serum of MMP-13 compared with the model group, there was a decrease in the high dose group,Chinese medicine, was much lower than western medicine group, Chinese medicine dose group and high dose group curative effect is better than that of traditional Chinese medicine, suggests the presence of a concentration dependent, inter group comparison, P<0.01, the difference was statistically significant.Conclusion:Knee osteoarthritis rat model induced by iodoacetic acid has a short cycle, high repetition rate and good stability. Short term use of Duhuojisheng decoction can reduce the expression of TNF-α, MMP-13 in articular cartilage tissues, inhibit the degradation of cartilage matrix, and promote the repair of articular cartilage, delay the degeneration of articular cartilage.It shows a dose dependent,the higher the dose, the stronger the inhibition. With wide indications, less side effect, obvious curative effect, suitable for long-term use,it is worthy of clinical promotion. |
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