ABSTRACT
BACKGROUND:Previous studies have found that electroacupuncture can delay articular cartilage degeneration mediated by JAK-STAT signaling pathway through upregulating the expression level of transforming growth factor β1 as well as mRNA expression levels of STAT3, Smad3 and LepR. In the meanwhile, electroacupuncture can inhibit the mRNA expression of p38 and Fas mRNA mediated by MAPK signaling pathways, further inhibiting the apoptosis of chondrocytes. OBJECTIVE: To explore the effect of electroacupuncture on the degeneration of articular cartilage in rats with knee osteoarthritis based on Ras-Raf-MEK1/2-ERK1/2 signaling pathway. METHODS:120 male healthy Sprague-Dawley rats aged 2 months olds were selected and randomly divided into normal, model, 15-minite electroacupuncture and 30-minute electroacupuncture groups (n=30 per group). The rats in the latter three groups received the intra-articular injection of 4% papain bilaterally, and the remaining rats received no intervention. At 2 weeks after modeling, the latter two groups were respectively given 15- and 30-minute electroacupuncture, five times weekly for consecutive 12 weeks. The morphology of the cartilage was observed by hematoxylin-eosin staining, the expression level of interleukin-1β in the synovium was detected by ELISA assay, and the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were detected by western blot analysis. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that: in the model group, the cartilage surface was rough, the cartilage layer became thinner, and the cartilage structure was damaged with incomplete tidal line; in the 15- and 30-minute electroacupuncture groups, the cartilage structure was complete with clear layers and complete tidal line. ELISA showed that the expression level of interleukin-1β in the model group was significantly higher than that in the normal group (P< 0.01), and the level in the 15- and 30-minute electroacupuncture groups was significantly lower than that in the model group (P < 0.05). Western blot assay found that compared with the normal group, the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were increased in the model group. However, all above protein levels except ERK1/2 in the 15- and 30-minute electroacupuncture groups were significantly lower than those in the model group (P < 0.01,P < 0.05). To conclude, electroacupuncture inhibits the degeneration of articular cartilage in osteoarthritisvia Ras-Raf-MEK1/2-ERK1/2 signaling pathway and downregulating the expression level of interleukin-1β.
ABSTRACT
BACKGROUND:Previous studies have showed thatTougu XiaotongCapsule (TGXTC) exertsbetter effects on osteoarthritis, byregulatingRho/Rock signaling pathway, inhibitingsignal transduction of chondrocyte mitochondrial apoptosis pathway,varyingthe rate and pattern of subchondral bone remodeling and improving the arrangement of subchondral bone colagen fibers and calcium-phosphate crystalization. OBJECTIVE:To observe the effects of the serum containing TGXTC and itsdisassembled recipeson chondrocytedegenerationof ratsviaWnt/β-cateninsignal pathway, and to explore the maintherapeutic method forosteoarthritis in theTGXTC. METHODS:FortySprague-Dawley rats were randomlyassigned to receivethe treatment ofTGXTC,Bushen Rougan(BSRG),Huoxue Qufeng(HXQF) and normal saline, respectively, according tothe dose conversion methods ofanimaltoanimal and animaltohuman. Thenvarious drug-containing serums wereprepared for thefolowingcelular experiment.After culture and passage, chondrocytesfromSprague-Dawley ratsat passage 3 were divided into five groups: blank control, model, TGXTC, BSRG, HXQF groups. Cels in the latter four groups wereculturedin appropriate drug-containing serums(normal salineserumfor the model group) for 72 hours, folowing intervention with interleukin-1β for 24 hours.Cels in the blank control group were cultured innormal saline serum.Afterwards, cels in al the five groups were colected for detecting expression ofWnt 4, β-cateninandmatrix metaloproteinase 13at mRNA and proteinlevels using real-time PCR and western blot assay, respectively. RESULTS AND CONCLUSION:Compared with theblank control group, the expressionof Wnt 4,β-catenin, matrix metaloproteinase 13 wassignificantly increasedin the model group. Compared with the model group, the expression of Wnt 4, β-catenin, matrix metaloproteinase 13 in the TGXTC, BSRG and HXQF groups were decreasedsignificantly, sequenced as TGXTC group