| Objective: Osteoarthritis (OA), also known as Osteoarthrosis, is a joint disease which is characterized by the degradation of articular cartilages and the formation of new bones at the joint margins, i.e., osteophytes. Knee OA, the most common form of OA, behaves as one of the main factors to cause the knee pain. The incidence rate is highly related to ages and makes knee OA the leading factor to affect movements and to result in chronic disables for senior people. Therefore more and more researchers have shown increasing interests in finding secure and highly effective treatments. This thesis is to establish the rats model of knee OA, and subsequently to observe the histological change of the knee and the effects of moxibustion therapy on the expression of TGF-β1, TGF-β3 and PTG. The presented investigation can demonstrate a powerful experimental evidence to support the clinical treatment.Methods: 39 2-month-old Wistar rats, weighted 175-200g, were randomly divided into 3 groups, i.e., normal group(A), model group(B) and moxibustion group(C), with 13 rats in each. General anesthesias were performed to all rats in the group B and C with a dose of 4ml Chloralhydrate (10%) per kg per rat. Knee OA was then induced by intra-articular injections with ling MIA diluted in 50μ1 physiological saline at the right knee and with 50μ1 physiological saline at the left as the intra-articular contrast, respectively. After 4 weeks, one randomly selected rat in each group was sacrificed with the knee joints subsequently stained by the hematoxylineosin. The knee cartilage histological changes were observed in line with the knee OA, which justifies the successful induction of the knee OA's. The treatments with moxibustion on the rats of group C were carried out from the 5th week. Lighted moxa-stick was used to perform moxibustion for Xiyan at 2- 3 cm off the skin for 20 minutes. Such a treatment was performed once a day for 3 periods in total each of which lasts 10 days. A break for one day was required between two consecutive periods. An inhalational anesthesia was conducted to the rats by mixing 5% isoflurane with air before the treatments, group A and B received no treatments.After the last treatment with the fasting for 24 hours, we sacrificed all the rats. The whole knee joints were taken out. Half of the samples were decalcified in 5% formic acid and processed for paraffin embedding. Subsequently, knee joints were dissected and the tissue sections (7μm) were placed on glass slides for staining. The other half of samples were placed in RPMI medium/0.1%BSA, incubated for 10 hours.The immunohistochemistry (ABC) was applied to detect TGF-β1, -β3; The contents of TGF-β1 and-β3 were measured by ELISA (Enzyme-linked Immunoadsordent Assay). The size of osteophytes and the PTG content in cartilages were measured by using the image analysis of histology.Results: 1. The surface area of osteophytes was significantly increased in the knee cartilages of rats' group B compared with group A (P＜0.01) . 2. The expression of PTG was significantly decreased in the knee cartilages of rats' group B compared with group A (P＜0.01). 3. The expressions of TGF-β1, TGF-β3 were significantly increased in the knee cartilages of rats' group B compared with group A (TGF-β1: P＜0.05, TGF-β3: P＜0.01).4. The surface area of osteophytes was significantly decreased in the knee cartilages of rats' group C compared with group B (P＜0.01) .5. The expression of PTG was significantly increased in the knee cartilages of rats' group C compared with group B (P＜0.01)6. The expressions of TGF-β1, TGF-β3 were significantly decreasedin the knee cartilages of rats' group C compared with group B (TGF-β1: P＜0.05, TGF-β3:P＜0.01) .Conclusion: The results indicated that the rats' model of experimental knee OA can be successfully established by injecting MIA into the knee joints. The moxibustion treatment on the Xiyan acupoint can improve the cartilage degeneration and reduce the osteophyte formation of the experimental knee OA. The moxibustion treatment of knee OA reduces the osteophyte formation in the knee cartilage for the experimental knee OA, most likely by down-regulating the high expression levels of TGF-β1, TGF-β3 in the knee cartilage of knee OA, which favors the PTG synthesis or inhibits the PTG degradation in the chondrocytes. |
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