| BackgroundOsteoarthritis is a common chronic degenerative disease in the elderlypopulation. The main features were move joint degeneration and mild disorders,accompanied by progressive damage to the articular cartilage. Although the etiologywas different, they had similar biology, morphology and clinical manifestations.Among them, the hip and knee were the most likely site of involvement, leading tochronic joint pain, dysfunction, and even lost the ability. It is serious harmful to thequality of life in the elderly population and gave the family and society heavy losses.Currently, the clinical treatment of osteoarthritis mainly based on first-line drugs, butthe long-term treatment would bring a series of drawbacks, and even worse.Therefore, finding effective and less side effects of therapy was particularly important.In this study, second-line therapy of glucosamine hydrochloride and curcumin werecombined to treat osteoarthritis of the knee by using animal specimens, and the cartilage cell apoptosis and apoptosis related genes were discussed.ObjectiveThe purpose of this study was to build rabbits knee osteo arthritis model andstudy osteoarthritis of the knee cartilage cell apoptosis, to analysis the impact ofglucosamine hydrochloride and glucosamine hydrochloride combined with curcuminon knee joint cartilage apoptosis, to discuss the influence of glucosaminehydrochloride and glucosamine hydrochloride combined with curcumin on knee jointcartilage of apoptosis-related proteins of Bcl-2, Bax and P53, and also, to furtheranalyze the effects of glucosamine hydrochloride and glucosamine hydrochloridecombined with curcumin on the knee joint cartilage of apoptosis-related genes ofBcl-2, Bax and P53in the genetic level.Methods40rabbits were randomly divided into control group (group A), model group(group B), glucosamine hydrochloride treatment group (group C), glucosaminehydrochloride combined with curcumin treatment group (group D). Each group had10rabbits. Modified Hulth method was used to copy knee arthritis animal model in B,C and D group. A group was used as a control group. After models built four weeks,C group was administered daily150mg/kg glucosamine hydrochloride, D group wascarried out within9mg/kg5%curcumin knee injections per week on the basis of Cgroup, A group and B group were given intra-articular injection of saline. Four groupswere continuously for4weeks. After four weeks, the experimental animals werekilled, the knee cartilage specimens of four groups models were taken. Knee cartilagecell apoptosis of the models in four groups were detected by using terminaldeoxynucleotidyl transferase-mediated in situ end translation method. Bcl-2, Bax andP53protein expression were detected in knee cartilage by usingimmunohistochemical methods. Bcl-2, Bax and P53gene expression levels inchondrocytes of the four group models were analyzed by RT-PCR experiments.Results1. Animal model built situation:40rabbits were completed throughout theexperiment, half-death situation did not appear,30animal models of osteoarthritiswere successfully established. 2. Cartilage cell apoptosis situation of animal models: The differences of kneecartilage apoptosis rates in the four group models is of statistically significant(P<0.05). By LSD-t test between any two groups, the AI values of B group, C groupand D group were higher than A group (P<0.05). The AI values of C group and Dgroup were lower than B group (P<0.05). The AI value of D group was lower than Cgroup (P<0.05).3. Cartilage cell apoptosis-related protein expression situation in animal models:The differences of Bcl-2protein expressing positive cell rates of knee cartilage cellsin the four group models were statistically significant (P<0.05). By using LSD-t test,pairwise differences among the four groups were statistically significant (P<0.05). Bgroup, C group and D group were lower than A group. C group and D group werehigher than B group; D group was higher than C group. The differences of Baxprotein expressing positive cell rates of knee cartilage cells in the four group modelswere statistically significant (P<0.05). By LSD-t test between any two groups, Bgroup, C group and D group were higher than A group (P<0.05). C group and Dgroup were lower than B group (P<0.05). D group was lower than C group. Thedifferences of P53protein expressing positive cell rates of knee cartilage cells in thefour group models were statistically significant (P<0.05). By using LSD-t test,pairwise differences among the four groups were statistically significant (P<0.05). Bgroup, C group and D group were higher than A group (P<0.05). C group and Dgroup were lower than B group (P<0.05). D group was lower than C group (P<0.05).4. Cartilage cell apoptosis-related gene expression situation in animal models:Bcl-2, Bax and P53gene expressions were detected by RT-PCR. The differences ofBcl-2gene expressions of knee cartilage cells in the four group models werestatistically significant (P<0.05). By LSD-t test between any two groups, theexpression levels of B group, C group and D group were lower than A group (P<0.05).The expression levels of C group and D group were higher than B group, D groupwas higher than C group, the differences were all statistically significant (P<0.05).The differences of Bax gene expressions of knee cartilage cells in the four groupmodels were statistically significant (P<0.05). By LSD-t test between any two groups,the expression levels of B group, C group and D group were higher than A group (P<0.05). C group and D group were lower than B group (P<0.05). D group waslower than C group (P<0.05). The differences of P53gene expressions of kneecartilage cells in the four group models were statistically significant (P<0.05). ByLSD-t test between any two groups, B group, C group and D group were higher thanA group (P<0.05). C group and D group were lower than B group (P<0.05). D groupwas lower than C group (P<0.05).Conclusions1. Rabbits knee osteo arthritis models could be successfully replicated bymodified Hulth method.2. Articular cartilage cells were undergoing apoptosis in rabbit model ofosteoarthritis of the knee. The incidences of apoptotic cells were much higher thancontrol group which did not occur arthritis.3. Articular chondrocytes apoptosis-related proteins, such as Bcl-2, Bax and P53,in rabbit models of osteoarthritis of the knee had changed. Bcl-2protein expressionpositive rate was lower than the control group, Bax protein expression rate was higherthan the control group, and P53positive expression rate was higher than the controlgroup, the differences were all statistically significant (P<0.05).4. By RT-PCR technique, Bcl-2gene expression in rabbits knee arthritis modelarticular cartilage cells was lower than the control group, Bax gene expression wasincreased compared with the control group, and P53gene expression was increasedcompared with the control group, the differences were all statistically significant(P<0.05).5. Glucosamine hydrochloride could be effectively raised Bcl-2protein levels inrabbits knee arthritis model of articular chondrocytes, reduced Bax protein and P53protein expression levels. And thus, it could inhibit apoptosis of articular cartilage.The combined effects of curcumin were much higher than the single use ofglucosamine hydrochloride.6. Glucosamine hydrochloride could enhance the level of Bcl-2gene expressionin rabbit articular chondrocytes knee arthritis model, reduce Bax and P53geneexpression levels, thereby inhibit apoptosis in articular chondrocytes. The effect ofcombining with curcumin was enhanced, which was stronger than glucosamine hydrochloride alone. |
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