The Effect Of Electroacupuncture At ST-36 On Murine Gastrointestinal Macrophages And Its Possible Mechanisms

Part 1 Electroacupuncture ameliorates the acute colitis via regulating lamina propria macrophages and its mechanismsAims Electroacupuncture(EA)at ST-36 can attenuate acute experimental colitis,but the mechanisms are unclear.We investigated the role of colonic lamina propria macrophages(L_pM_?)in the anti-inflammatory effects of EA and its molecular mechanisms.Methods Male C57BL/6 mice were randomized into five groups: normal control,dextran sulfate sodium(DSS)-induced acute colitis(DSS),DSS with sham EA(SEA),DSS with high-frequency EA(HEA)and DSS with low-frequency EA(LEA).Body weight,colon length,DAI score and histological score were evaluated during colitis progression.Serum and colonic levels of pro-and anti-inflammatory cytokines were detected with ELISA,cytometric beads array,RT-PCR and western blotting analysis.Colonic L_pM_? subsets were determined using flow cytometry.Magnetic-activated cell sorting was applied to isolate colonic L_pM_?,and molecular mechanisms were explored with western blotting,RT-PCR and immunofluorescence.Results(1)Compared with the DSS group,HEA and LEA attenuated body weight loss and decreased DAI and histological scores.(2)Serum levels and colonic protein and m RNA levels of IL-1?,TNF?,IL-6,IL-12 and IL17 were markedly decreased with HEA and LEA.IL-10 level was increased with HEA.(3)M1 percentage increased,while M2 percentage decreased in the DSS group;HEA and LEA reversed these proportions.(4)NLRP3/IL-1? protein and m RNA levels in isolated colonic L_pM_? decreased with HEA and LEA compared with the DSS treatment group;HEA increased Nrf2/HO-1 levels compared with levels in DSS mice.Conclusions Theanti-inflammatory effects of EA on DSS-induced acute colitis may rely on regulating L_pM_? polarization,NLRP3/IL-1? suppression and Nrf2/HO-1 promotion.Part 2 Electroacupuncture promotes gastrointestinal motility via regulating muscularis macrophages and its mechanismsAims Electroacupuncture(EA)at ST-36 could accelerate the delayed gastrointestinal(GI)motility in many GI motility dysfunction models,but the definite effect and mechanisms are unclear.Recently,the vital role of muscularis macrophages(MM_?)in regulating GI motility has been varified.In this study,We intended to investigate the effects of EA on intestinal manipulation(IM)mice model and involved mechanisms.Methods Male C57BL/6 mice were randomized into five groups: normal control,intestinal manipulation(IM),IM with sham EA(SEA),IM with high-frequency EA(HEA)and IM with low-frequency EA(LEA).The GI transit was evaluated.The infiltration of MM? and its phenotype were analyzed with flow cytometry.Magnetic-activated cell sorting was applied to isolate MM?,and the relationship between the MM_? and interstitial cells of Cajal(ICCs)was further investigatedResults(1)Compared with the IM group,HEA and LEA attenuated the delayed intestinal transit.(2)Both the HEA and LEA obviously reduced the MM? and suppressed the M1 activation of the MM? in the ileum.(3)EA restored the disrupted ICC networks through inhibiting the release of IL6 by the MM_?.Conclusions(1)EA at acupoint ST-36 could accelerate the delayed intestinal transit in the IM murine model.(2)EA could restore the disrupted ICC networks in IM model.(3)EA reduced the infiltration of MM_?,inhibited its M1 polarization and IL6 secretion and restored the ICC networks,resulting the recovery of delayed intestinal transit in IM model.