GSK3B Overexpression Alleviates Posttraumatic Osteoarthritis in Mice by Promoting DNMT1-Mediated Hypermethylation of NR4A3 Promoter

Figure 6

GSK3B promotes NR4A3 promoter methylation by recruiting DNMT1 in vivo. C57BL/6 mice were randomly divided into 5 groups: sham, posttraumatic oleanolic acid (PTOA)+vector, PTOA+adeno-related virus GSK3B overexpression vector (Ad-GSK3B), PTOA+scramble, and PTOA+sh-NR4A3, 10 mice per group. Destabilization of medial meniscus (DMM) surgery was performed on mice to generate a posttraumatic OA model. Immediately after the surgery, the joint was injected with 100 μL of the following adeno-related virus vectors (empty vector, Ad-GSK3B, scrambled shRNA, and sh-NR4A3), respectively, and the virus titer was . (a) Safranin O/fast green staining was used to assess cartilage damage in the sham, PTOA+vector, and PTOA+Ad-GSK3B groups. (b) Mouse OARSI score. (c–e) Western blotting was used to detect the expression of GSK3B and NR4A3 proteins in mouse knee joint tissues. (f) The change of DNMT1 bind to the promoter of NR4A3 by chromatin immunoprecipitation (Ch-IP) method. Data is presented as the and analyzed by Student’s -test or ANOVA test. , and .

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