Experimental Study On Achyranthes Bidentata Against Articular Cartilage Influence Of OA Models Of Rabbits

ObjectiveObserved the pathophysiological changes of the knee joint’s articular cartilage of osteoarthritis model of rabbit, to further explore the nosogenesis of osteoarthritis and analysis the Achyranthes bidentata mechanism of action of OA rabbit knee cartilage, to provide some basis for experimental research and clinical application in field of Chinese medicine prevention and treatment of osteoarthritis.MethodsNew Zealand white rabbits were randomly divided into normal group, model group, treatment groups (Achyranthes bidentata) and the control group (Celebrex). The treatment groups were divided into Achyranthes bidentata high dose group, Achyranthes bidentata middle dose group and Achyranthes bidentata low dose group. Reference to the Hulth’s Modeling method to make the experimental rabbit knee OA model. Harvested the rabbits with macth dose Achyranthes bidentata after modeling one week, six weeks of continuous treatment, detected the changes of morphology and pathophysiology of lesions cartilage tissue, specifically divided into four parts: 1Use the Chemical Preparation of cartilage tissue slices, to observed lesions of cartilage tissue morphological changes observed in the light microscope and electron microscope.2Extracted the synovial fluid of knee joint, to detect the content of NOS and SOD by the chemical colorimetric assay.3Use the Western blot method to detect the expression of MMP-3, MMP-13and TIMP-1in synovial tissue. Compared the deference of each group by image analysis system.4The use of immunohistochemical techniques were used to detect the expression of cartilage tissue’s IGF-1and type Ⅱ collagen. Compared the deference of each group by image analysis system.Results1Histological changes:the normal group of the articular cartilage surface was smooth, evenly distributed chondrocytes; cartilage tissue surface of the model group was significantly thinner, showing that the cells clustered chondrocytes derangement; Achyranthes bidentata high dose group, Achyranthes bidentata medium dose group and control group’s cartilage tissue damage were reduced significantly, a few regions visible cartilage cells arranged in irregular; Achyranthes bidentata low-dose group had little difference with the model group. Cartilage tissue score:the model group’s score increased than the normal group (P<0.01). The score of Achyranthes bidentata high dose group, medium dose group and control group were significantly lower than the model group (P<0.01). There were no significant difference between the medium dose group and high dose group (P>0.05), but the two groups both had differences compared with the control group (P<0.01). No significant differences between Achyranthes bidentata low dose group and the model group (P>0.05). Electron microscope obse rvation:the cartilage cells of the normal group were normal; the model group’s cartilage cells had irregular shape. There were condensation phenomenon in cartilage tissue; Achyranthes bidentata high dose group and medium dose group,the chondrocyte had Mildly abnormal. Part of the chondrocyte had some vacuoles. The degree of cell damage was better than the control group and Achyranthes bidentata low dose group.2NOS values in the synovial fluid of model group had significantly increased than in normal group (P<0.01). Achyranthes bidentata high dose group, Achyranthes bidentata medium dose group and control group were significantly lower than the model group (P<0.01). NOS values of the control group and Achyranthes bidentata high dose group were lower than NOS values of Achyranthes bidentata medium dose group (P<0.01). Achyranthes bidentata low dose group had no difference with model group (P>0.05). SOD values in the synovial fluid of model group had significantly decreased than in normal group(P<0.01). Achyranthes bidentata high dose group SOD levels was higher than the medium dose group, and the two groups had significantly increased than the model group (P<0.01).There were no difference between Achyranthes bidentata low dose group、 control group and model group (P>0.05)3The expression of MMP-3、MMP-13in synovial tissue had no difference between Achyranthes bidentata high dose group、 Achyranthes bidentata medium dose group and control group (P>0.05). The three groups were lower than model group (P<0.01). The expression of TIMP-1in synovial tissue of Achyranthes bidentata high dose group and Achyranthes bidentata medium dose group and control group were higher than model group (P<0.01). The expression of TIMP-1in synovial tissue of Achyranthes bidentata high dose group and Achyranthes bidentata medium dose group were higher than the control group (P<0.05). The expression of MMP-3、MMP-13and TIMP-1in synovial tissue had no difference between Achyranthes bidentata low dose group and model group (P>0.05)4IGF-1expression of Achyranthes bidentata high dose group was higher than the middle dose group (P<0.01), and the two groups were significantly higher than control group、the Achyranthes bidentata low dose group and model group (P<0.01). Expression of collagen Ⅱ, Achyranthes high-dose group and medium dose group control group and model group were higher than model group(P<0.01). Achyranthes high-dose group and medium dose group were higher than control group (P<0.05). Achyranthes low-dose group had no difference with model group (P>0.05)ConclusionsA certain dose of Achyranthes bidentata can delay the degener ation of cartilage tissue in the OA model; Achyranthes bidentata probably by altering the OA lesion cartilage tissue’s NOS and SOD activity, to protect the cartilage tissue and delay the progression of the disease; Achyranthes bidentata can inhibit the expression of MMPs and increase TIMP-1expression to control the OA disease development. The effective concentration of Achyranthes bidentata can through the way of increase expression of IGF-1and type Ⅱ collagen protein in OA cartilage tissue to delay the progression of the disease.