电针刺激大肠俞及足三里穴对肝缺血再灌注大鼠肠道屏障功能的影响
2021-07-27王成琛魏来谭思由邹毅孔高茵
王成琛 魏来 谭思由 邹毅 孔高茵



〔摘要〕 目的 探究術前电针(electric acupuncture, EA)刺激大肠俞及足三里穴,是否对肝缺血再灌注(ischemia-reperfusion, I/R)大鼠的肠道屏障功能具有保护作用。方法 32只SD大鼠随机分为假手术组(Sham组)、缺血再灌注组(I/R组)、穴位刺激缺血再灌注组(AS+I/R组)、非穴位刺激缺血再灌注组(nAS+I/R组);于肝脏缺血1 h再灌注4 h后分别取其下腔静脉血及小肠标本,使用血清学方法检测丙氨酸转氨酶(alanine aminotransferase, ALT)、天冬氨酸转氨酶(aspartate aminotransferase, AST)浓度,Chiu's法评价HE染色后光镜下肠黏膜损伤程度,ELISA法检测血清D-乳酸(D-lactate, D-Lac)、二胺氧化酶(diamine oxidase, DAO)水平,免疫组化检测小肠组织中闭锁小带(ZO-1 tight junction, ZO-1)蛋白表达情况。结果 与Sham组比较,I/R组大鼠AST、ALT浓度较高,肠黏膜损伤严重,血清D-lac、DAO浓度较高,ZO-1蛋白表达较低(P<0.05);与I/R组和nAS+I/R组比较,AS+I/R组大鼠Chiu's评分较低,血清AST、ALT浓度较低,血清D-Lac、DAO水平较低,ZO-1蛋白表达较高(P<0.05);且与I/R组比较,nAS+I/R组血清AST、ALT浓度和血清DAO水平较低,ZO-1蛋白表达较高(P<0.05)。结论 EA刺激大肠俞及足三里穴,能减轻肝I/R大鼠的肠道损伤,减少菌群入血,对其肠道的屏障功能具有保护作用。
〔关键词〕 肝缺血再灌注;肠道屏障功能;电针;大肠俞;足三里
〔中图分类号〕R245 〔文献标志码〕A 〔文章编号〕doi:10.3969/j.issn.1674-070X.2021.03.010
〔Abstract〕 Objective To explore whether electroacupuncture (EA) stimulation on Dachangshu (BL25) and Zusanli (ST36) acupoints before surgery has protective effects on intestinal barrier function in hepatic ischemia reperfusion (I/R) rats. Methods 32 SD rats were randomly divided into the sham operation group (Sham group), ischemia-reperfusion group (I/R group), acupoint-stimulated ischemia reperfusion group (AS+I/R group) and non-acupoint-stimulated ischemia reperfusion group (nAS+I/R group). After 1 hour of ischemia and 4 hours of reperfusion, inferior vena cava blood specimens and small intestine samples of rats were taken respectively, serological methods were used to detect alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations, Chius evaluation method was used to evaluate the degree of intestinal mucosal damage under microscope after HE staining, ELISA method was used to detect serum D-lacticate (D-Lac) and diamine oxidase (DAO), immunohistochemistry was used to detect the expression of ZO-1 tight junction (ZO-1) protein in small intestine. Results Compared with the Sham group, the concentrations of AST and ALT in I/R group were higher, the intestinal mucosa injury was serious, the concentrations of serum D-lac and DAO were higher, and the expression of ZO-1 protein was lower (P<0.05); compared with I/R group and nAS+I/R group, the Chiu's score, the concentrations of serum AST and ALT, the levels of serum D-lac and DAO were lower, and the expression of ZO-1 protein was higher in AS+I/R group (P<0.05); and compared with I/R group, the concentrations of serum AST and ALT, the level of serum DAO were lower, the expression of ZO-1 protein was higher in nAS+I/R group (P<0.05). Conclusion The stimulation of EA on Dachangshu (BL25) and Zusanli (ST36) points can alleviate the intestinal damage of hepatic I/R rats, reduce the influx of bacteria into the blood, and protect the intestinal barrier function.
〔Keywords〕 hepatic ischemia reperfusion; intestinal barrier function; electroacupuncture; Dachangshu (BL25); Zusanli (ST36)
肝脏手术常需暂时性阻断入肝血流,而重新恢复血液供应后可能造成肝缺血再灌注损伤(hepatic ischemia-reperfusion injury, HIRI)[1-3]。这种损伤不仅仅局限于肝脏本身,也可能对全身其他脏器功能造成影响[4],导致围术期一系列不良反应及并发症,肠道是缺血再灌注(ischemia-reperfusion, I/R)损伤最常累及的器官。穴位刺激(acupoint stimulation, AS)是中医的重要组成部分,电针(electric acupuncture, EA)作为目前临床上常用的AS方法,通过对神经、内分泌、免疫等系统的调控,对多个器官和系统产生保护作用[5-6]。在现有的针对胃肠保护行AS的动物实验及临床研究中,大肠俞和足三里穴选用较多,功能效果较为明确。本实验拟探究肝脏手术前EA刺激大肠俞及足三里穴对肝I/R大鼠肠道的屏障功能的影响,为AS保护肠道功能运用于临床提供理论依据。……
