电针"天枢""上巨虚"调控NF-κB通路改善腹泻型肠易激综合征结肠低度炎症的机制研究OA

Objective To observe the effects of electroacupuncture at Tianshu(ST25)and Shangjuxu(ST37)on long non-coding RNA NEAT1(lncRNA NEAT1),tumour necrosis factor receptor superfamily member 1B(TNFRSF1B),and nuclear factor-κB(NF-κB)in a irritable bowel syndrome with diarrhea(IBS-D)rat model,thereby elucidating the specific mechanisms underlying electroacupuncture treatment for IBS-D.Methods 53 newborn SD rat pups were randomly assigned to control group(with 12 pups)and modelling group(with 41 pups).Pups in modelling group underwent maternal separation,colonic instillation of acetic acid,and chronic restraint stress to establish the IBS-D model.Ultimately,36 successfully modeled pups were selected and randomly assigned to the model group,inhibitor group,and electroacupuncture group,with 12 rats in each group.The inhibitor group received intraperitoneal injections of the NF-κB inhibitor PDTC(50 mg/kg).The electroacupuncture group underwent bilateral electroacupuncture stimulation at the Tianshu(ST25)and Shangjuxu(ST37),administered once daily for 7 consecutive days.Diarrhea rate and colorectal distension pain threshold(AWR score)at baseline,post-modeling,and post-intervention in each group were compared.After the intervention,colon tissue samples from each group were collected for hematoxylin and eosin(HE)staining.Serum levels of TNF-α,IL-1β,and IL-8 were measured using enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of NEAT1,TNFRSF1B,and NF-κB p65 in colon tissues were determined by qRT-PCR.Western blot was performed to assess the protein expression of TNFRSF1B and NF-κB p65 in colon tissues.Immunofluorescence staining was used to detect the positive expression of NF-κB p65,TNF-α,IL-1β,IL-8,and TNFRSF1B in colonic tissue.Results Compared with the control group,rats in the model group exhibited a significantly reduced AWR scores(P<0.01),and markedly increased diarrhea rate(P<0.01).The colonic mucosa exhibited inflammatory infiltration along with structural disorganization.Serum levels of the inflammatory cytokines TNF-α,IL-1β,and IL-8 increased(P<0.01).The mRNA expression of NEAT1,TNFRSF1B,and NF-κB p65 in colon tissues was increased(P<0.01),protein expression of NF-κB p65,TNF-α,IL-1β,IL-8,and TNFRSF1B increased(P<0.01),the positive expression of NF-κB p65、TNF-α、IL-1β、IL-8 and TNFRSF1B increased(P<0.01).Compared with the model group,rats in the inhibitor and electroacupuncture groups exhibited significantly elevated AWR scores(P<0.01)and markedly reduced diarrhea rates(P<0.01).Colonic tissue morphology remained relatively intact,with diminished inflammatory responses and reduced inflammatory cell infiltration.Serum levels of TNF-α,IL-1β,and IL-8 decreased(P<0.01),and the mRNA expression of NEAT1,TNFRSF1B,and NF-κB p65 in colon tissues decreased(P<0.01).The positive expression of NF-κB p65,TNF-α,IL-1β,IL-8,and TNFRSF1B in colon tissues decreased(P<0.01).Conclusion Electroacupuncture stimulation at Tianshu(ST25)and Shangjuxu(ST37)significantly alleviates diarrhoea and visceral hypersensitivity symptoms in IBS-D rats.Its therapeutic mechanism may involve suppressing lncRNA NEAT1 expression,blocking TNFRSF1B/NF-κB signaling pathway activation,thereby reducing the production of core inflammatory mediators and ameliorating colonic low-grade inflammation.