| Objective: In this study,SD rats were selected as the research objects to observe the impact of GuBiZhiTong(GBZT)solution on BMP2 and Sox9 expression in the articular cartilage of knee osteoarthritis in rats through the animal experiment and to explore the protective impact of GBZT solution on the articular cartilage of knee osteoarthritis.Methods: 45 male and 45 female SPF SD rats of 280+/-20 g weight were selected.15 of them were randomly selected as control group.The rest of them were established the models of knee osteoarthritis by using improved Hulth method.After moding,they were randomly divided into model group,low,medium,high does groups of GBZT solution and glucosamine hydrochloride(GH)group.15 rats were included in each group,and male and female rats were kept in separate cages.After 8 weeks' continuous drug intervention,rats in each group were put into death by the neck off method and sampled articular cartilage specimen.Naked-eye observation was performed for morphology changes and pathological sections of hematoxylin-eosin(HE)HE staining were observed by microscope.Real-time polymerase chain reaction(RT-PCR)method was taken to detect the expression of BMP2 mRNA and Sox9 mRNA in cartilages.Results:(1)Macroscopic observation:the cartilage in the control group were the same with that in normal's.The joint surface was smooth,the appearance and color of the cartilage articularits were normal,articular surface was transparent and complete.Joint surface ulcers,subchondral bone exposure or hyper-plastic change didn't occur.The model group joint surface was rough and not smooth,articular cartilage faded dull and had a lower transparency,erosion,exfoliation and defect occurred,osteophyte was formed in the edge of joint,synovium was thicken and subchondral bone exposed.The transparency of cartilage in low dose group,medium dose group,high dose group and GH group was higher than that in the model group with less smoothness and darkcolor in some area.There was erosion,a part cartilage defect and little osteophyte in low dose group,but not in medium dose group,high dose group and GH group.(2)HE staining :Mankin's criteria was referred to compare pathological sections of HE staining.Compare the model group with the control group.In the control group,the cartilage surface was smooth,the dyeing was cleat,the surface layer,Transition layer,radiation layer and calcified layer of cartilage were clearly discernible,cells ranked neatly,levels were clear,chromatin distributed equally and the nucleus could be clearly seen.In the model group,the cartilage surface was rough,the surface layer became thin or even disappeared.Some areas were fluffy,cracks could reach deeper,and pycnosis and necrosis appeared in some of the nucleus.The calcified layer cells arranged in disorder and formed cluster.formed cell cluster.Tidal lines were incomplete or disappeared.There was statistically significance(P < 0.05)in the two groups.The cartilage damages in each dose,GBZT solution groups and the GH group were better than that in the model group but worse than that in the control group.There were still some areas of cartilage cells arranged in irregular phenomenon,but obviously better than that in the model group with less clear boundary layers,microscopic phenomenon of individual regions had family collection.Dyeing matrix was uniform,and individual tidal lines were incomplete.There was statistically significance(P<0.05)in the two groups.(3)PCR detection :compared with the moded group,the BMP2 and Sox9 mRNA expression in rats cartilage specimen in the low and medium dose groups was increased,but there was no statistical significance(P>0.05).The BMP2 mRNA and Sox9 mRNA expression in the high dose group and the GH group was significantly increased(P<0.05).Compared with the low and medium dose groups,the expression of BMP2 and Sox9 mRNA increased obviously(P<0.05).Conclusions:(1)GBZT solution can raise the expression of BMP2 and Sox9 in knee joint cartilage in the rat model of KOA and impact the pathological changes of OAarticular cartilage,thus reducing joint damage,protecting the articular cartilage and delaying the progress of OA disease.(2)High doses of GBZT solution has a more significant clinical curative effect on KOA. |
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