Effect Of Bushen Tongluo Prescription On Cartilage Cell Microfilament Structure Osteoarthritis Model In Rats

ObjectiveConstruction of SD rat knee osteoarthritis model by modified Hulth method, to observe the effect of Bushen Tongluo Prescription on the model of rat knee cartilage cytoskeleton microfilament structure, and then explore the molecular mechanism of the Bushen Tongluo prescription in the treatment of osteoarthritis of the knee.MethodsFrom animal breeding center, 60 SD rats were randomly divided into four groups, which include four groups sham JS(Sham-operated) group, MX(model) group,XY(medicine, ibuprofen) group, ZY(TCM) group, each group has 15. Four groups,MX group, XY group, ZY group‘s models were made by using modified Hulth method,(surgical approach: transection of rats on the right knee medial collateral ligament SD,anterior cruciate ligament, a complete separation of the medial meniscus board), rats of JS group only cut the joint capsule. Six weeks after conventional breeding, the establishment of knee osteoarthritis animal model to X-ray Meets osteoarthritis performance modeling to determine whether the successful, start of the administration to determine the first day after the model was successful. Each dose group: JS Group,MX group were given normal saline(1ml / 100g) orally; XY group received ibuprofen suspension(1ml / 100g) orally; ZY group was given traditional Chinese medicine(1ml/ 100g) irrigation stomach; the frequency of administration of each group were once a day gavage for 6 weeks. Given medicines to six weeks, all experimental SD rats were sacrificed, take rat right femoral condylar Gross observation. After the general observation for all the right femoral condyle regular decalcification, paraffin embedding and sectioning. The above sections were HE staining on the right side of the right femur condylar cartilage morphology. Meanwhile modified Mankin's score of cartilage tissue were evaluated. Take the above sections using immunofluorescence cartilage actin cytoskeleton fluorescence staining using Confocal Laser Scanning Microscope(CLSM) to detect actin morphology and fluorescence intensity.Results(1).Gross observation: JS group within the right lateral femoral condyle cartilage surface is smooth and no degeneration. MX group within the right lateral femoral condyle cartilage degeneration serious. The XY group the right lateral femoral condyle cartilage degeneration obvious, but compared to the outside with the inside of the lighter. ZY of right medial femoral condyle cartilage mild degeneration of the lateral condyle smooth without degeneration.(2).cartilage Histological observation: JS group within the lateral femoral condyle cartilage four complete structure, the number of normal chondrocytes. Four-part structure of the lateral condyle cartilage in the area were damaged femur MX group, the number of chondrocytes decreased, less damage to the inside. XY Group medial femoral condyle cartilage damage partial region layer of mast cells, reducing the number of cells, the lateral condyle cartilage injury to the intermediate layer, increasing the number of cells. ZY group medial femoral condyle cartilage surface damage, cartilage cells in normal quantities, the lateral condyle cartilage with JS Group is no different.(3).observation of actin: in the lateral femoral condyle cartilage JS group of actin fibers and a linear shape parallel to the long axis of the cell. MX Group within the lateral femoral condyle cartilage microfilaments occurrence of significant shrinkage. XY Group medial femoral condyle cartilage microfilaria occurrence of significant shrinkage, lateral cartilage thickening and actin moderate shrinkage, but the degree than the inside of the light. ZY group within the lateral femoral condyle cartilage actin and JS compared maintained good linear form.(4).Mankin's Rating: comparison between groups: within the lateral femoral condyle cartilage four groups of Mankin's score compares four groups are not all equal(p<0.01); respectively, within among the four groups lateral condyle cartilage Mankin's scores two Compare the two and found differences in four groups in the medial condyle of any difference between the two groups were statistically significant, the lateral condyle of any two groups were statistically significant. Trends within the lateral condyle of the four groups of Mankin's score performance, as supreme MX group, followed by the group XY, ZY group than in the XY group but higher than JS group. Within the group: In addition to the differences in the JS Group and MX lateral condyle of Mankin's score was not statistically significant(p> 0.05), Mankin's score was in the XY and ZY lateral condyle of the group was statistically significant(p<0.01).(5).Quantitative analysis of actin: Four groups were the lateral femoral condyle skeletal actin fluorescence quantitative analysis of cartilage cells, actin content difference between the groups in the lateral condyle were not statistically significant(p> 0.05).Conclusion(1)There are morphological changes in actin filament of the cytoskeleton of the articular cartilage of osteoarthritis;(2) Bushen Tongluo Fang can inhibit osteoarthritis of the knee joint cartilage cell actin filament structure deformation and protect the cartilage;(3) It is not change that the content of normal and osteoarthritis of the actin filament of the cartilage cells.