Intervention effect of Tuina pressing and kneading the Huantiao (GB30) acupoint on NF-κB p65 protein at spinal cord dorsal horn in sciatica rats / 中国骨伤

OBJECTIVE@#To observe the analgesic effect of Tuina by pressing and kneading the Huantiao (GB30) acupoint on rats with chronic constriction injury (CCI) and to explore the analgesic mechanism of Tuina on sciatica rats.@*METHODS@#Thirty-two SPF male SD rats weighing 180 to 220 g were randomly divided into fore groups:blank group (without any treatment), sham group (only exposed without sciatic nerve ligating), model group (sciatic nerve ligating) and Tuina group (manual intervention after lsciatic nerve ligating). The CCI model was prepared by ligating the right sciatic nerve of the rats, on the third day of modeling, the rats in the Tuina group were given pressing and kneading the Huantiao (GB30) point for 14 days, and the changes of paw withdrawal threshold(PWT), paw withdrawal latency(PWL) were measured before and on the 1st, 3rd, 7th, 10th, 14th and 17th days after modeling. The changes of sciatic functional index(SFI) were measured before and on the 1st and 17th day after modeling. The morphological changes of the sciatic nerve were observed by hematoxylin-eosin(HE) staining;and the differences in NF-κB protein expression in the right dorsal horn of the spinal cord of rats were detected.@*RESULTS@#Following modeling, there was no significant difference in PWT, PWL and SFI between the blank group and the sham group (P>0.05), but the PWT, PWL and SFI of the model group and the Tuina group decreased significantly (P<0.01). After manual intervention, the pain threshold of rats in Tuina group increased. On the 8th day of manual intervention (the 10th day after modeling), PWT in Tuina group increased significantly compared with that in model group (P<0.01). On the 5th day of manual intervention (the 7th day after modeling), the PWL of the massage group was significantly higher than that of the model group (P<0.01). The pain threshold of rats in Tuina group continued to rise with the continuous manipulation intervention. After 14 days of manipulative intervention, the sciatic nerve function index of rats in the Tuina group increased significantly(P<0.01). Compared with the blank group and sham group, the myelinated nerve fibers of sciatic nerve in the model group were disordered and the density of axons and myelin sheath was uneven. Compared with the model group, the nerve fibers of rats in the Tuina group were gradually continuous and the axons and myelin sheath were more uniform than those in the model group. Compared with the blank group and sham group, the expression of NF-κB protein in the right spinal dorsal horn of the model group was significantly increased(P<0.01). Compared with the model group, the expression of NF-κB protein in the right spinal dorsal horn of rats in Tuina group decreased significantly(P<0.01).@*CONCLUSION@#Pressing and kneading the Huantiao (GB30) point restores nerve fiber alignment;and improves the PWT、PWL and SFI in the CCI model by decreasing NF-κB p65 protein expression in the spinal dorsal horn. There fore, Tuina demmstrates an analgesic effect and improves the gait of rats with sciatica.

Effects and mechanism of β-carotene on inflammatory factors in LPS-induced RAW264.7 cells / 中国免疫学杂志

Objective:To investigate the effects and mechanism of β-carotene on inflammatory factors (IL-1 β,IL-6,TNF-α) in LPS-induced RAW264.7 cells.Methods:Firstly,RAW264.7 cells of being induced by 4 (5 μg/ml)for 24 h were treated with different concentration of β-carotene (20,40,80,160 pmol/L)for 3 h.The cells viability was measured by MTIT,the mRNA relative expression of IL-1 β,IL-6,TNF-cα was detected by fluorescence quantitative PCR,the secretion capacity of IL-1 β,IL-6,TNF-α was detected by ELISA and the protein relative expression of NF-κB p65 protein was measured by Western blot.Secondly,RAW264.7 cells were induced by LPS(5 μg/ml) and different concentration of PDTC(1,5,10 μg/ml)for 24 h,NF-κB p65 protein was measured by Western blot and inflammatory factors were detected by fluorescence quantitative PCR and ELISA.Finally,compared the changes in the relative expression of inflammatory factors and NF-κB p65 protein between LPS+PDTC group and LPS+PDTC + β-carotene group.Results:Compared with the LPS-induced group,β-carotene could increase the cell viability of LPS-induced RAW264.7 cells and inhibied the relative expression of inflammatory factors and NF-κB p65 protein.Inhibited the relative expression of NF-κB p65 protein could reduce the relative expression of inflammatory factors.Compared with the LPS+PDTC group,LPS +PDTC + β-carotene group could inhibit the relative expression of inflammatory factors significantly (P＜0.05).But,there was little difference about the relative expression of NF-κB p65 protein between this two groups.Conclusion:β-carotene inhibits the relative expression of inflammatory factors(IL-1 β,IL-6,TNF-α) in LPS-induced RAW264.7 cells through inhibition of NF-κB p65 protein in NF-κB pathway,this pathway isn't unique.