| ObjectiverTo observe the impact of digital-pressing manipulation at "Dazhui" point with spoon needle on the inflammatory cytokines and the expression of P-MKK3/6^ P-p38MAPK on the MKK3/6-p38MAPK signal transduction pathways in chondrocytes of rats with cervicovertebral disc degeneration.Methods:The 40 SD rats (male and female half) were randomly divided into a sham group (n=10) and a model group (n=30). The sham group were only given the skin incision from the back of the neck and then sutured, while the model of intervertebral disc degeneration was established by using static-dynamic force unbalance in the model group. When the model was confirmed successfully by picro-sirius red (PSR) staining 3 months later, the rats in which were randomly divided into three groups:model control group (control group), model medication administration group (MA group) and model digital-pressing manipulation group (DPM group), with 8 rats for each group. In the digital-pressing manipulation group, the Tekscan’s Flexiforce ELF System pressure measuring joint were put on the "Dazhui" point of the rats, while a professional man handed a spoon needle like holding a pen, and put it on the pressure measuring joint then applied digital-pressing manipulation. Technique of power was measured by FlexiForce ELF System. The power of stimulus was controlled in the (4±0.5) N. After the needle applied digital-pressing manipulation on the point was about five seconds, the man who raised his hand and the needle three seconds digital-pressed the point as before.The time of treatment was 20 minutes each time,1 time each day, consecutive 14 days for one course, a total of 2 courses. A second course can be implemented after the first one was completed for 2 days. The MA group was given to the meloxicam tablets lavage treatment, one time each day, the same treatment time and courses as the DPM group. The sham group and the model group were only fixed in the same conditions without any treatment. After the end of the courses the rats from each group were killed. Immunohistochemistry and western blotting were used to detect the expression of TNF-a IL-1β and P-MKK3/6, P-p38MAPK in the disc cartilage cells from each group.Results:(1) Picro-sirius red staining:Compared with the sham group, the model group showed type I collagen incerased and type Ⅱ collagen decreased obviously while their proportion in the intervertebral disc type collagen system happened a significant change.(2) Immunohistochemistry (IHC):Compared with the sham group, the contents of TNF-α and IL-1β in the model group were increased significantly (P<0.05). Compared with the control group, the contents of TNF-α and IL-1β in the DPM group and the MA group were decreased significantly (P<0.05).(3) Western Blot:Compared with the sham group, the contents of P-MKK3/6 and P-p38MAPK in the model group were increased significantly (P<0.05). Compared with the control group, the contents of P-MKK3/6 and P-p38MAPK in the DPM group and the MA group were decreased significantly (P<0.05).Conclusion:(1) Digital-pressing manipulation at "Dazhui" with spoon needle could reduce the contents of TNF-α, IL-1β in chondrocytes of rats with cervical intervertebral disc degeneration.(2) Digital-pressing manipulation at "Dazhui" with spoon needle could decrease the expression of the P-MKK3/6、 P-p38MAPK in the disc cartilages regulation.(3) Digital-pressing manipulation at "Dazhui" with spoon needle could reduce the expression of the TNF-α, IL-1β in chondrocytes possibly through regulation of MKK3/6-p38MAPK signaling pathway, so as to reduce the inflammatory reaction in chondrocytes to delay the degeneration of cervical intervertebral disc. |
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