| Objective：To investigate the effect of Fuyuan capsule(FYC)on articularcartilage histomorphology，to study the effect of Fuyuan capsule on theexpression of a disintegrin and metalloproteinase with thrombospondinmotifs、marix metalloproteinase and Transforming Growth Factor-β1inarticular cartilage of the rat knee osteoarthritic(OA)model.To DiscussFuyuan capsule on osteoarthritis cartilage function mechanism of theprotection.Methods：180male adult SD rats were divided into normal group, model group,Kanggu zengsheng capsule group and fuyuan capsule high,medium andlow dose group.Except the normal control group,the others were inducedby Hulth’s method to copy model of KOA.A week later,the treatmentgroups were given different drugs for4、8and12weeks, while the modeland normal control groups were prefuse with the same amout ofphysiological saline.samples of joint were collected at the4th、8thand12th week. histomorphology of articular cartilage be observed by lightmicroscopy and naked-eye. ADAMTs-4，ADAMTs-5，TGF-β1，MMP-1and MMP-3were determined by Immunohistochemical technology and theexpression of ADAMTs-4、 ADAMTs-5and TGF-β1mRNA wereevaluated by RT-PCR technology.The amout of protein expression ofcollagen type Ⅱ、MMP-1、MMP-3、ADAMTs-4and ADAMTs-5weredetected by Western blot assay.Results1. The effects of Fuyuan capsule on histomorphology of articular cartilagein knee OA ratsThe articular cartilage of normal group shows the normal morphology.After model establishment, at4weeks, the articular cartilage of modelgroup Begin to appear cartilage surface erosion, cracks and otherpathological changes. At8weeks of OA cartilage surface damage isaggravating, heaviest at12weeks cartilage surface damage. After the drugtreatment, Fuyuan capsule and Kanggu zengsheng capsule degree of OAarticular cartilage pathological changes were reduced, and the articularcartilage surface damage of Fuyuan capsule high dose group was obviouslyrelieved.2. Effects of Fuyuan capsule on ADAMTs-4and ADAMTs-5in articularcartilage of knee OA ratsImmunochistochemistry test showed that at4weeks, compared with normal group, expression of ADAMTs-4and ADAMTs-5in the modelgroup were Increased significantly (p <0.01), Dyeing is tan; comparedwith model group,expression of ADAMTs4and ADAMTs5in Fuyuancapsule groups on rat articular cartilage were lower significantly, therewere significant differences (p <0.01)；compared with Kanggu zengshenggroup,expression of ADAMTs4and ADAMTs5in Fuyuan capsule groupson rat articular cartilage were lower significantly（p <0.05）RT-PCR test showed that the expression of ADAMTs-4and ADAMTs-5mRNA in Normal group of articular cartilage was low, at4weeks,compared with normal group, expression level of mRNA were increasedsignificantly in model group of articular cartilage, to higher mRNAexpression for the ADAMTs-5,(p <0.01); ADAMTs-4and ADAMTs-5mRNA expression of Fuyuan capsule group were reduced in Differentlevels, compared with model group, there were statistically significantdifferences in Fuyuan capsule high dose group (p <0.05, p <0.01)；ADAMTs-4and ADAMTs-5mRNA expression of Fuyuan capsule inmiddle and high dose group were reduced in Different levels, comparedwith Kanggu zengsheng group（p <0.05）。Western blot assay show: protein expression of ADAMTs-4andADAMTs-5were increased Compared with normal group,obviously inmodel group on articular cartilage; compared with model group, Theexpression of ADAMTs-4and ADAMTs-5were reduced in Fuyuan capsule high dose group; And reduce the amount of the ADAMTs-5proteinexpression range is more obviously（p <0.01）；compared with Kangguzengsheng group, The expression of ADAMTs-4and ADAMTs-5werereduced in Fuyuan capsule high dose group（p <0.05）。3.Effects of Fuyuan capsule on collagen type Ⅱ、MMP-1and MMP-3inarticular cartilage of knee OA ratsImmunochistochemistry test showed that at4、8and12weeks, modelgroup were the positive expression of MMP1MMP-3particles atfull-thickness cartilage, is located in the cytoplasm, Dyeing is tan. theexpression of MMP-1and MMP-3in Fuyuan capsule groups is lower thanthe model group (p <0.05), and at12weeks, the expression of MMP1andMMP-3in Fuyuan capsule high dose group is significantly lower than themodel group, the difference is statistically significant (p <0.01)；Theexpression of MMP1and MMP-3in Fuyuan capsule high dose group issignificantly lower than the Kanggu zengsheng group (p <0.05).Western blot assay showed that expression of MMP-1and MMP-3inNormal group is only a small amount, while the expression of MMP-1andMMP-3in model group is higher than normal group, with increased proteinexpression of MMP-3more obvious(p <0.01); the expression of MMP-1and MMP-3in Fuyuan capsule high dose group is significantly lower thanthe model group.(p <0.01)； the expression of MMP-1and MMP-3inFuyuan capsule high dose group is significantly lower than the Kanggu zengsheng group.(p <0.05)。Western blot assay showed that the content of type II collagen in thearticular cartilage of Normal group is in rich. Compared with normal group,the content of type II collagen on articular cartilage is decreasedsignificantly in model group, and increasing over time (p <0.01).Compared with model group, the content of type II collagen is increased inFuyuan capsule high dose group; there was statistically significantdifference (p <0.01)4. Effects of Fuyuan capsule on TGF-β1in articular cartilage of knee OAratsImmunochistochemistry test showed that the expression of TGF-β1isvisible on full-thickness cartilage cells within the cytoplasm and nuclei ofall in Normal group.at4、8and12weeks, the expression of TGF-β1isreduced significantly on articular cartilage in model group, and the moreserious with the increase of time. the expression of TGF-β1in Fuyuancapsule group is higher than model group, which the expression of TGF-β1are highest in Fuyuan capsule high dose group (p <0.01)；the expressionof TGF-β1in Fuyuan capsule high dose group is higher than the Kangguzengsheng group (p <0.05)。RT-PCR test showed that the mRNA expression of TGF-β1in thearticular cartilage of Normal group is in rich. Compared with normal group,the mRNA expression of TGF-β1on articular cartilage is decreased significantly in model group, and increasing over time (p <0.01).compared with model group, the expression of TGF-β1is increased inFuyuan capsule high dose group,there was statistically significantdifference (p <0.01);compared with Kanggu zengsheng group, theexpression of TGF-β1is increased in Fuyuan capsule high dose group(p <0.05)。Conclusions：1. Fuyuan capsule can improve general grade and Mankin’s score onarticular cartilage of rat knee OA, which established by Hulths method,have protective effect on cartilage degeneration.2. Fuyuan capsule can inhibit the expression of ADAMTS-4andADAMTS-5, reduced the degradation of proteoglycan from reaching effecton prevention and treatment of Early osteoarthritis.3. Fuyuan capsule can significantly improve the expression of TGF-β1inknee joint cartilage of rats, can inhibit the expression of mmp-1andmmp-3,reduced the degradation of collagen type II from reaching effect onprevention and treatment of osteoarthritis. |
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