电针调节IL-10/DDiT4/mTOR信号通路改善大鼠溃疡性结肠炎的机制研究OA

Objective To investigate the potential mechanism by which electroacupuncture(EA)alleviates intestinal inflammation in rats with ulcerative colitis(UC)via modulating the interleukin-10(IL-10)/DNA damage-inducible transcript 4(DDiT4)/mammalian target of rapamycin(mTOR)signaling pathway and regulating colonic cell autophagy.Methods Thirty-two healthy male Sprague-Dawley(SD)rats aged 6-8 weeks were randomly divided into a normal control group,a UC model group,an EA group,and a Western medicine group,with 8 rats in each group.The UC model was induced by allowing rats free access to drinking water containing 5%dex-tran sulfate sodium(DSS)for 7 days.Rats in the EA group received EA at the bilateral"Tianshu"(ST 25),"Shangjuxu"(ST 37),and"Zhongwan"(CV 12)acupoints,stimulated with dense-sparse wave(10 Hz/50 Hz alternately)for 20 minutes daily for 3 days.Rats in the Western medicine group were administered mesalazine suspension(200 mg/kg·bw)by gavage once daily for 3 days.The disease activity index(DAI)was evaluated.hematoxylin-eosin(HE)staining,Alcian blue-periodic acid-Schiff(AB-PAS)staining,and trans-mission electron microscopy(TEM)were used to observe colonic pathology,mucus secretion,and ultrastructure,respectively.Serum levels of IL-10,IL-6,tumor necrosis factor-α(TNF-α),and interleukin-1β(IL-1β)were measured by enzyme-linked immunosorbent assay(ELISA).The protein expression levels of IL-10,DDiT4,mTOR,and the autophagy-related proteins LC3B,ATG5,and p62 in colonic tissues were detected by Western blot(WB).The mRNA expression levels of IL-10,DDiT4,and mTOR in colonic tissues were measured by quantitative reverse transcription polymerase chain reaction(RT-qPCR).Results Compared with the normal control group,the DAI level in the UC model group rats was increased(P<0.01),the content of anti-inflammatory factor IL-10 decreased(P<0.01),the contents of pro-inflammatory factors IL-6,TNF-α,and IL-1βincreased(P<0.01),the expressions of IL-10,DDIt4,and LC3Ⅱ/LC3Ⅰ,and ATG5 in the colonic tissue decreased(P<0.01),the expressions of mTOR and p62 increased(P<0.01),the mRNA expressions of IL-10 and DDiT4 decreased(P<0.01),and the mRNA expression of mTOR increased(P<0.01).Compared with the UC model group,the DAIs in the EA group and Western medicine group decreased(P<0.01),the content of IL-10 increased(P<0.01),the contents of IL-6,TNF-α,and IL-1β decreased(P<0.01),the expressions of IL-10,DDIt4,LC3 Ⅱ/LC3 Ⅰ,and ATG5 increased(P<0.01),the expressions of mTOR and p62 decreased(P<0.01),the mRNA expressions of IL-10 and DDiT4 increased(P<0.01),and the mRNA expression of mTOR decreased(P<0.01).Conclusion EA may promote colonic epithelial cell autophagy and alleviate intestinal inflammation in rats with UC by regulating the IL-10/DDiT4/mTOR signaling pathway,providing a new perspective for UC treatment.