Experimental Study On Gene Expression Profile Of Early Knee Osteoarthritis Model In Rabbits By Moxiao Decoction

Objective:to study the miao medicine fumigation therapy on early experimental rabbit knee osteoarthritis of the research of gene expression profilesMethods:the rabbit knee joint cavity injection of papain copy early knee osteoarthritis model, then with miao medicine fumigation therapy after20days, with gross observation, under the light microscope, scanning electron microscopy (sem) to observe the pathological changes of cartilage and Mankin, s method;Using gene chip technology to detect gene expression of cartilage tissue, again to the original data obtained by gene chip detection after preprocessing and normalization correction processing, using relevant software for data difference gene (DEG) analysis, cluster analysis, gene function (GO) analysis and difference signaling pathways (Pathway) analysis.Results:(1) through general observation:model group:visible to change on the surface of the articular cartilage.(2) the blank control group: normal articular cartilage. HE dyed visible cartilage surface smooth, level off;Normal cartilage thickness, four layers structure clear, the tide line clear, complete structure;Normal cartilage cells, cartilage pit and tidy uniform distribution, sequence, cartilage matrix normal dyeing, alcian blue staining in dark blue. Model control group:articular cartilage for early cartilage damage. HE dyed visible cartilage surface rough, uneven;Levels close to normal cartilage thickness, four layers structure disorder, cartilage surface fibrosis, erosion, radiation layer and deep cartilage matrix fiber loose, visible to fat deposition in the matrix;Surface reduced Numbers of chondrocytes, deep the cluster cartilage cells can be increased and the disordered arrangement, normal cartilage pit disappear, tide line disappear or ambiguous, incomplete structure;Cartilage matrix staining, alcian blue staining in light blue. Miao medicine fumigation groups:articular cartilage damage. HE dyed visible cartilage surface slightly rough;Levels close to normal cartilage thickness, four layers structure clear, slightly fibrosis cartilage surface layer, radiation layer and deep normal cartilage matrix fibers;Cartilage cells was equal to the normal cartilage, cartilage cells arranged a little mess, pit normal cartilage;Tide line are clear and completeCartilage matrix dyeing slightly decreased, alcian blue staining in blue.Water fumigation groups:articular cartilage damage is a bit light. HE staining, cartilage surface rough, uneven;Thinning of the cartilage thickness, four layers structure level is a bit disorder, cartilage surface fibrosis, radiation layer and deep cartilage matrix fiber loose, visible to the subtle fat deposition in the matrix;Surface chondrocytes part number decreased, deep chondrocytes part straw cocooning frame set, cell arrangement is a bit messy, tide line slightly fuzzy is incomplete;Cartilage matrix staining mild decrease, alcian blue staining with pale blue.(3) groups of Mankin’s score results show that the model control group compared with blank control group with significant difference (P<0.01);Miao medicine fumigation group compared with model control group with significant difference (P<0.01);Water fumigation group compared with model control group with significant difference (P <0.01), miao medicine fumigation with water fumigation group compared with significant difference (P<0.01).(4) that can be observed under the scanning electron microscopy (sem):the blank control group:normal cartilage. Model control group:the cartilage surface uneven, cartilage surface of collagen fibers network structure disorder, collagen fiber surface there are a lot of floe coverage;Most of the collagen fiber in capsule sample, fiber porous structure between fuzzy, disordered arrangement of space. Collagen fiber, arranged disorderly, or even rupture even increases, insect damage sample change. Miao medicine fumigation groups:cartilage surface is flat, cartilage surface close to the normal articular cartilage surface, collagen fibers were a funicular, are intertwined with net. But collagen fiber surface a floc cover, slightly fuzzy between fiber porous structure. Owe water fumigation groups: cartilage surface level off, the cartilage surface of collagen fibers network structure disorder, collagen fiber surface there are a lot of floc coverage;Most of the collagen fiber in capsule sample, circular protrusions. The porous structure of fuzzy, space between fiber disordered arrangement. Some collagen fiber even show insect damage model change.(5)①DEG analysis:model control group relative to the control group genes were significantly differentially expressed827, up there435, down there392, Miao medicine fumigation group relative to the model group genes were significantly differentially expressed208, of which there are112regulated, there are96down, water fumigation group relative to the model control group were significantly differentially expressed genes are152, of which there are93up-regulated, down there59.②cluster analysis: samples of each experimental group can be divided into categories, each specimen reproducible experiments, each experimental group relative to the control group of gene expression has its own characteristics.③GO analysis:significant observation of208DEG GO classification, which relates to the classification of biological processes inflammation, cell proliferation, cell differentiation, protein, nucleotide, lipid metabolism, wherein the matrix metalloproteinase3(MMP3), scavenger receptor CD36(CD36) genes involved in several biological processes.④Pathway Analysis:The changing cartilage Miao medicine fumigation treatment involves a relatively early KOA important signaling pathways: Wnt signaling pathway, calcium signaling pathway, MAPK signaling pathway, osteoclast differentiation factor signaling pathways.Conclusion:This research through the light microscope, electron microscope, gene chip detection technology emergence medicine fumigation therapy can delay the articular cartilage degeneration of KOA, gene chip detection technology can effectively filter out with the onset of KOA differentially expressed genes, confirmed that the onset of KOA is interaction of complex process that multiple genes through further analysis of these genes, you can find the key genes miao medicine fumigation therapy in the treatment of KOA, for the prevention and treatment of KOA provide new targets and ideas of development.