| Objective Osteoarthritis is the most common joint diseasedepartment of orthopedics field, with the rapid development ofChina’s aging process, the incidence of OA will continue to rise,seriously affecting the quality of life in the elderly. The mainpathological changes of osteoarthritis of articular cartilage indifferent degree of degeneration or disappearance, and subchondralbone destruction. Pathological study confirmed, joint cartilagedegeneration of osteoarthritis is the characteristic pathologicalchanges, pathological changes of OA is the most basic. The cartilagedegeneration mechanism and the cellular and molecular basis ofbecome a research hotspot in recent years. The subject of the useof Cynanchum paniculatum paeonol effect on matrixmetalloproteinase as the breakthrough point, to explore themechanism of Radix Cynanchi paniculati Danpi treatment ofosteoarthritis from three aspects of cell morphology,ultrastructure changes under the electron microscope, molecularbiology, and provide theoretical and experimental basis for theclinical application of Radix Cynanchi paniculati paeonol for osteoarthritis of the knee.Methods40weight2kg (±100g) healthy adult New Zealand whiterabbits, irrespective of gender, were randomly divided into normalgroup, control group, saline group, triamcinolone acetonide group,Radix Cynanchi paniculati paeonol group, with8rats in each group.Randomly selected8rats as normal group, the remaining32rabbitmedial meniscus of knee joint before1/3hepatectomy andtransection of the anterior cruciate ligament (ACLT) preparationof osteoarthritis (OA) model method of [4] modeling,4weeks Houzaomodel. Normal group: without any treatment. Control group: only therabbit knee medial meniscus of1/3hepatectomy and transectionof the anterior cruciate ligament (ACLT) preparation ofosteoarthritis (OA) model method of molding, no other treatment.The physiological saline group, Maria Nade group, Radix Cynanchipaniculati paeonol groups respectively: group right kneeintra-articular injection of0.9%saline, Maria Nade, Xu ChangqingDanpi0.2ml began modeling after first weeks,2times/week, a totalof10times, for5weeks. All animal were completed5weeks aftertreatment, the ear vein injection amount were exposed to air, kneejoint cavity. The naked eye observation of articular cartilagesurface smoothness, luster, surface erosion, whether there is afibrous hyperplasia and subchondral bone exposure morphologicalchanges, take pictures and score according to the general scorestandard; and the prepared the medial femoral condyle cartilagespecimens, transmission electron microscope ultrastructure wereobserved and photographed.②from the right medial femoralcondyle articular surface of about2mm thickness of articularcartilage were made into paraffin sections, two step immunohistochemical staining under light microscope, theorganizational structure of the detailed observation, and integralstatistics Mankin score according to Mankin articular cartilagepathology score standard, package structure includes jointcartilage, changes in cell number, tidal line of complete and easydyeing four aspects; statistical analysis of articular cartilagematrix metalloproteinases (MMP-1) and its inhibitor (TIMP-1)positive index expression. The use of MMP-1and TIMP-1oligonucleotide probe by RNA in situ hybridization method, as thelocalization and quantification of rabbit articular cartilage ofMMP-1and TIMP-1mRNA, detected the expression of rabbit kneeosteoarthritis cartilage tissue MMP-1and TIMP-1mRNA。Results①gross observation showed the normal articularsurface was smooth and intact. The control group cartilage changeis concentrated in the medial femoral condyle, articular cartilagehas eroded and stripping of cartilage defect. Saline groupcartilage surface rough, hypertrophy. Cynanchum paniculatumpaeonol articular surface was slightly rough, mild cartilageerosion. Triamcinolone acetonide group cartilage erosion degreethan Cynanchum paniculatum weight group, some of the subchondralbone exposure. Electron microscope showed that the normal cells ofnormal morphology, nuclear, cytoplasmic density uniform, note theamount of organelles, cell surface villous and collagen clearlyvisible. The control group most cartilage cells are irregular inshape, cell organs in cytoplasm dissolved, collagen arranged indisorder. Saline group most cartilage cell swelling. Cynanchumpaniculatum group: cell morphology remained mild disorder,collagen, rough endoplasmic reticulum rich. Triamcinolone acetonide group: granular appearance, cytoplasmic organellesdissolved, less collagen fiber content.The examination of normalgroup Mankin score0-1points in the organization; Cynanchumpaniculatum paeonol group with mild to moderate injury, Mankinscore of4-6; triamcinolone acetonide group with medium cartilageinjury, Mankin score of8-10; model group with severe cartilageinjury, Mankin score11-12points; saline group was similar to modelgroup. Cynanchum paniculatum paeonol group cartilage tissue MMP-1expression positive index higher than the normal control group, theMaria Nade group and model group, there was significant differencebetween groups (P <0.05); the expression of Cynanchum paniculatumpaeonol group cartilage tissue TIMP-1level is Maria Nade groupincreased, compared between the two groups had significantdifference (P <0.05), the Maria Nade group the expression levelof TIMP-1and model group, saline group comparisons between groupsshowed no significant difference (P>0.05). The normal articularcartilage of MMP-1and TIMP-1mRNA expression was low or noexpression; expression of MMP-1and TIMP-1mRNA Cynanchumpaniculatum paeonol group cartilage tissue were lower; increase theexpression of triamcinolone acetonide group cartilage MMP-1mRNA;the model of articular cartilage strongest MMP-1mRNA expression,TIMP-1mRNA layer no expression or low expression of Cynanchumpaniculatum; paeonol group cartilage tissue MMP-1mRNA levelshigher than the normal control group, compared with triamcinoloneacetonide group low, with a significant difference between groups(P <0.05); Xu Qing expression of cartilage tissue of paeonol groupTIMP-1mRNA levels were triameinolone de group, there wasstatistically significant difference between two groups (P <0.05). Conclusions these results suggest that expression of jointdamage and MMP-1positive correlation, Cynanchum paniculatum Danpicould inhibit the expression of MMP-1, the elimination of MMP-1incartilage of excessive extracellular matrix degradation. At thesame time, the MMP-1specific inhibitor TIMP-1with MMP-1secretionincreases also increased, but not in sync with the increase of MMP-1,the secretion level of their difference, may make the MMP-1excessive play on the degradation of articular cartilage.Expression of joint damage and TIMP-1are negatively correlated,Cynanchum paniculatum paeonol could promote the expression ofTIMP-1, and thus inhibit the expression of MMP-1function.Therapeutic effect of Radix Cynanchi paniculati Danpi onosteoarthritis is through inhibition of MMP-1mRNA expression incartilage injury and promote the expression of TIMP-1mRNA of thebidirectional regulation mechanism to achieve. |
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