| Objective: To observe the influence of GuBiZhiTong(GBZT) solution oninterleukin(IL)-1,tumor necrosis factor(TNF)-αin articular cells of experimental animalmodels through establishing the New Zealand white rabbits model of knee osteoarthritis.Toexplore the mechanism of the treatment of osteoarthritis and provide theoretical basis for thetreatment of osteoarthritis.Methods: Fifty male or female healthy New Zealand white rabbits of2.5kg to3kgweight were randomly divided into5groups.There were low,medium,high dose groups ofGBZT solution, glucosamine hydrochloride(GH) group and model group respectively. And10rabbits were included in each group. Hulth method was refered to establish the model ofosteoarthritis. After moding, different dose of GBZT solution, glucosamine hydrochloride andnormal saling(NS) were intragastric administration for each group continuously for eightweeks.Rabbits in each group were put into death and sampled articular cartilage specimenafter modling for four weeks and eight weeks respectively. Naked-eye observation wasperformed for morphology changes and pathological sections of hematoxylin-eosin(HE)staining were observed by microscope.Real-time polymerase chain reaction(RT PCR) methodwas taken to detect the expression of IL-1and TNF-α in rabbit knee cartilagearticularis.The statistical analysis was performed by SPSS19.0software.Results:1. Macroscopic observation: Normal knee joint surface should be smooth andhas transparent color. After4weeks’ moding, the model group articular cartilage faded dull,joint surface was not smooth, partly was beige, no obvious defects and the synovium wasthicken. The transparency of cartilage in medium dose group, high dose group and GH groupwas higher than that in the model group and without defects and new creatures. After8weeks,the articular cartilage became yellow in the model group and the transparency of cartilage wassignificantly reduced. There were also osteophyte significantly formed and the cartilage evenbecame erosion. But there no obviously changes in the GBZT solution groups and GH groupcompared with the previous groups. significant And involving the bone pain liquid group andglucosamine hydrochloride group, no obvious changes earlier.2. HE staining microscopic observation:The cartilage issue surface became thin and even partially disappeared in the model group. Some areas were fluffy and had rough surface,showed some of the cracks or exfoliation. Uneven distribution of cartilage cells destiny wasobserved. There was cell clustering phenomenon exists and no significant cartilage cellsarranged in order. Most of the tidal lines were not complete. The cartilage damages in the lowdose, medium dose, high dose GBZT solution groups and the GH group were better thanthat in the model group. There were still some areas of cartilage cells arranged in irregularphenomenon, but obviously better than that in the model group with less clear boundarylayers, microscopic phenomenon of individual regions had family collection. The majority ofthe tidal lines were imcomplete but individual integrity. Structural damage of the cartilage andthe formation of the osteophyte were observed in the model group after8weeks. There wereno different in each treatment group compared with before.3. RT-PCR: Compared with the model group after8weeks, the expression of IL-1mRNA and TNF-αmRNA in the cartilage specimens of the white rabbits were significantlower in the low dose of GBZT solution group(P<0.05). The expression of IL-1mRNA andTNF-αmRNA were significant lower in the medium dose, high dose and the GH groupscompared with the model group(P<0.01).The expression of IL-1mRNA and TNF-αmRNAwere significant lower in the medium dose, high dose of GBZT solution groups and the GHgroup compared with the low dose of GBZT solution group after8weeks(P<0.05).Comparedwith the GH group after8weeks, there were no significant different of expression of IL-1mRNA and TNF-αmRNA in the medium dose, high dose of GBZT solution groups(P>0.05).The expression of IL-1mRNA and TNF-α mRNA in the low dose, medium dose,high dose of GBZT solution groups and the GH group after8weeks were significantly lowerthan that of after4weeks.Conclusion: The GBZT solution could effectively delay the degeneration of kneecartilage. The GBZT solution showed a better regulation and control of the expression of IL-1and TNF-α factors in the cartilage cells. The mechanism might be that the GBZT solutioncould reduce the expressions of pro-inflammatory cytokines(IL-1,TNF-α) and inhibit theinflammatory response so as to play a role on anti-inflammatory and thereby reliving jointdamage. This can provide the basis for the clinical use of the GBZT solution. The treatment ofGBZT solution on knee osteoarthritis reflect the advantages of traditional Chinese medicine in the treatment of difficult diseases and provide new researches and directions for clinicalresearch of OA. |
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