Experimental Study On The Effects Of Point-pressing Manipulation With Spoon Needle On NF-κB P50,NF-κB P65 And Inflammatory Cytokines In Chondrocytes Of Rats With Cervicovertebral Disc Degeneration

Objective:To observe the effect of digital-pressing manipulation at "Dazhui" point with spoon needle on the nuclear factor kappa-B p50(NF-κB p50),nuclear factor kappa-B p65(NF-κB p65),Interleukin-la(IL-la)and Interleukin-17(IL-17)inflammatory cytokines in chondrocytes of rats with cervicovertebral disc degeneration,and explore the function of them in the process of intervertebral disc degeneration.Methods:40 healthy SD rats were randomly divided into sham group(n=10,male and female half),model group(n=30,male and female half).The model of rats was established degeneration of cervical intervertebral disc by using static-dynamic imbalance method.The sham group was only cutting the skin incision and made with the intervention of skin incision suture,not cutting off the muscle tissue.The model was confirmed successfully establishment by toluidine blue staining after 3 months of conventional breeding.The rats of model group were randomly divided into model control group(control group),model digital-pressing manipulation group(DPM group)and model meloxicam treatment group(MT group),8 model rats for each group.Using self-made elastic clamped DPM group of rats,the method of digital-pressing manipulation is that,the Flexiforce ELF System pressure measuring joint were put on the "Dazhui" point of the rats,while a professional man handed the spoon needle like holding a pen,and put it on the pressure measuring joint then applied digital-pressing manipulation.The power of stimulus was controlled in(4±0.5)N,continued for 5 seconds,two intervals for 3 seconds.The time of treatment was 20 minutes each time,1 time each day,consecutive 14 days for one course,two courses between interval of 2 days,a total of 2 courses of treatment.The MT group was treated with meloxicam with doses of 0.7878mg/kg/d by gavage.Both of sham group and model group were olny fixed in the same conditions without any treatment.Take the intervertebral disc after the end of treatment.Immunohistochemistry was used to detect IL-la and IL-17 protein expression in the disc cartilage cells;Western blotting was used to detect NF-κB p50 and NF-κB p65 proteins expression in the disc cartilage cells.Results:(1)Toluidine blue staining:Cartilage endplate toluidine blue stain was present purple blue.The thickness of the endplate thicker and the number of endplate cartilage cell increased markedly in the sham group.Cartilage endplate toluidine blue stain in the control group faded.The thickness of the endplate and the number of endplate cartilage cell decreased markedly.(2)Immunohistochemistry(IHC):Compared with the sham group,the expression of IL-la and IL-17 proteins in the control group were increased considerably(P<0.05);Compared with the control group,the contents of IL-la and IL-17 proteins in the DPM group and MT group were decreased considerably(P<0.05).(3)Western Blot:Compared with the sham group,the expression of NF-κB p50 and NF-κB p65 proteins in the control group were increased considerably(P<0.05).Compared with the control group,the contents of NF-κB p50 and NF-κB p65 proteins in the DPM group and MT group were decreased considerably(P<0.05).Conclusion:(1)Digital-pressing manipulation at "Dazhui" with spoon needle could reduce the contents of IL-la and IL-17 in chondrocytes of rats with cervical intervertebral disc degeneration.(2)Digital-pressing manipulation at "Dazhui" with spoon needle could decrease the expression of the NF-κB p50 and NF-κB p65 in the disc cartilages regulation.(3)Digital-pressing manipulation at "Dazhui" with spoon needle delay disc degeneration may be through controling the expression of NF-κB and the inflammatory cytokines of IL-la and IL-17 in chondrocytes.