推拿调控NLRP3炎症小体通路对腰椎间盘突出症大鼠的抗炎镇痛效应OA

Objective:To observe the effects of Tuina Huantiao(GB30)on rats with lumbar disc herniation(LDH)and its anti-inflammatory and analgesic effect mechanism.Methods:A total of 20 clean-grade male SD rats were randomly divided into four groups:sham operation group,blank group,Tuina group,and model group,with 5 rats in each group.The model group was prepared by exposing the L4 and L5 intervertebral foramina through an incision in the right spinal skin and muscle and implanting a sterile L-shaped stainless steel rod to induce LDH.After successful modeling,the rats were housed for three days.On the fourth day,the Tuina group received Tuina therapy at the right Huantiao(GB30)for 15 minutes using a custom-made rat restraint,while the model group was restrained without Tuina therapy.The sham operation group had their skin and muscle incised and then sutured without any further intervention,and the blank group received no intervention.The changes in paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)were recorded in rats in each group 1 day before modelling and 4,6,8,10,13 and 17 days after surgery.After Tuina,HE staining was performed on the L4 and L5 intervertebral discs of each group to observe internal tissue changes.Protein expression levels of IL-1β,IL-18,NLRP3,ASC,and Caspase-1 in the L4 and L5 dorsal root ganglion(DRG)were detected using Western Blot analysis.Additionally,immunofluorescence(IF)was used to assess differences in NLRP3 activation levels in the L5 DRG across the groups.Results:On days 13 and 17 after post-surgery,the PWL and PWT in the Tuina group were significantly higher than the model group(P＜0.05,P＜0.01).Compared with the control group and the sham operation group,the model group and the Tuina group showed inflammatory cell invasion and degeneration in the lumbar intervertebral discs,Tuina was able to slow down degeneration and alleviate inflammation.Protein expression of IL-iβ,IL-18,NLRP3,ASC and Caspase-1 was significantly lower in the Tuina group than in the model group(P＜0.01,P＜0.05).The fluorescence intensity of NLRP3 in the model group was obviously higher than that in the blank and sham operation groups.After Tuina intervention,the fluorescence intensity of NLRP3 decreased obviously.Conclusion:Tuina therapy can inhibit the activation of the NLRP3 inflammasome in the DRG of LDH model rats and reduce the protein expression levels of downstream IL-1β and IL-18,thereby alleviating pain behavior and inflammatory responses in LDH rats.