| Objective:Through this experiment,we observed the expression of Wnt and ?-catenin in the articular cartilage of each group,and the effect on the signaling factor MMP13 downstream of the signal pathway.Provide a theoretical basis for the clinical application of therapeutic KOA.Method:1?The mold solution was prepared in advance: 2%(w/v)papain and 0.03 mol/L L-cysteine(2:1)mixed solution.Then,34 male New Zealand big ears rabbits were randomly divided into 9 blank groups and 25 model rats.After the rabbits were fixedly anesthetized,the right posterior knee joint cavity injection molding solution was performed on the first,third and fifth days,respectively,according to the injection dose of 0.1 mL/kg.After the model was fed for 2 weeks,a rabbit was randomly selected from the blank group and the model group.The rats were sacrificed after anesthesia,and the articular cartilage was taken for pathology.The cartilage degeneration was observed and the modeling was confirmed.l.2?After successful modeling,24 rabbits were divided into model group,control group and experimental group according to the random number table method,with 8 rats in each group.Then,each group was administered intra-articular injection 4 times on the 1st,4th,7th,and 10 th,the control group was injected with sodium hyaluronate injection,the experimental group was injected with aconite injection,and the model group was injected with normal saline..After continuing to raise for 1 week,All the 4 rabbits were sacrificed by air embolization,and the cartilage tissue was taken to observe the cartilage changes under HE staining;The expression of Wnt and ?-catenin in cartilage tissue was detected by immunohistochemistry;the cartilage tissue under light microscope was further observed;The mRNA expression of Wnt and ?-catenin in cartilage tissue was detected byRT-qPCR.The data were processed by statistical processing software SPSS19.0.The measurement data were expressed as((?)±s).One-way ANOVA was used for comparison between groups.P<0.05 was considered statistically significant.Result:1?Through experiments,the experimental specimens were observed.Compared with the model group and the control group,the articular cartilage was lightly damaged,thearticular cartilage was relatively intact,the cartilage surface crack was less,and the epiphysis was significantly reduced.2?Under light microscope,HE staining phase: the cytoplasm of chondrocytes in the experimental group(Wutou injection group)was pink and the nucleus was purple-blue.Compared with the model group and the control group,the cells showed no obvious deformation,vacuolar-like changes,no lobulation in the nucleus,and pyknosis.3?Immunohistochemical analysis showed that the expression of Wnt and ?-catenin in the experimental group Wuwutou injection group and the model group and the control group decreased(P<0.05),and the difference was statistically significant.Under the high power microscope,the nuclear staining of the experimental group and the control group showed a yellow distribution,which was significantly reduced.4?RT-qPCR:analysis: The mRNA expression of Wnt and ?-catenin in the articular cartilage of the experimental group(Wutou injection group)was lower than that of the model group and the blank group(P<0.01),and the difference was statistically significant and superior to the control group.Conclusion:Wutou injection can significantly down-regulate the expression of Wnt2,?-catenin and MMP-13 in articular cartilage.The mechanism of action is to reduce the damage to the articular cartilage by inhibiting the Wnt/?-catenin classical signaling pathway and treat KOA. |
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