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目的:探讨富氢生理盐水(HRS)对小肠缺血再灌注损伤(IIRI)大鼠肠黏膜屏障的影响。方法:将24只8周龄健康雄性SD大鼠按随机数字表法分为3组(每组8只)：假手术组、模型组和HRS组。HRS组大鼠在小肠缺血第30分钟时腹腔注射HRS(10 mL/kg)；模型组大鼠在相同时间点腹腔注射9 g/L盐水(10 mL/kg)。小肠缺血持续45 min，再灌注6 h后取材。采用酶联免疫吸附试验(ELISA)检测血清肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β和IL-17A水平；免疫组织化学染色(IHC)检测回肠黏膜闭合蛋白(Occludin)表达，免疫荧光(IF)检测黏连蛋白1(ZO-1)表达；Western blot检测回肠黏膜Occludin、ZO-1和Lysozyme蛋白表达，实时荧光PCR(qPCR)检测Lysozyme和α-defensin的mRNA表达。结果:ELISA结果显示：HRS组大鼠血清TNF-α与IL-1β水平较模型组明显降低[(62.02±29.97) ng/L比(113.40±44.58) ng/L，(21.68±0.35) ng/L比(28.29±3.49) ng/L ]；IL-17A水平升高[(28.18±5.28) ng/L比(15.10±3.60) ng/L ]，差异均有统计学意义(均n P<0.05)。IHC结果显示：与模型组大鼠相比，HRS组大鼠回肠黏膜Occludin染色较深，表达均匀连续。IF染色结果显示：与模型组大鼠相比，HRS组大鼠回肠黏膜ZO-1的荧光信号强度显著增加，分布连续清晰。Western blot结果显示：与模型组大鼠相比，HRS组大鼠回肠黏膜Occludin与ZO-1的蛋白表达明显增加(0.79±0.06比0.54±0.04，0.91±0.11比0.51±0.13)；Lysozyme的蛋白表达减少(1.50±0.40比2.99±0.80)，差异均有统计学意义(均n P<0.05)。qPCR结果显示：与模型组大鼠相比，HRS组大鼠Lysozyme的mRNA表达降低(1.64±0.33比2.20±0.40)；α-defensin的mRNA表达明显升高(0.82±0.19比0.47±0.13)，差异均有统计学意义(均n P<0.01)。n 结论:HRS通过维持肠黏膜屏障紧密连接蛋白Occludin和ZO-1的表达及调节抗菌肽α-defensin和Lysozyme的分泌减轻大鼠IIRI。“,”Objective:To investigate the effects of hydrogen rich-saline (HRS) on intestinal mucosal barrier in rat with intestinal ischemia/reperfusion injury (IIRI).Methods:Twenty-four healthy male Sprague-Dawley rats, aged 8 weeks, were randomly divided into 3 groups (8 in each group) by random number table method: sham group, model group and HRS group.Rats in HRS group were intraperitoneally injected with HRS (10 mL/kg) at 30 min of ischemia, and the same amount of normal saline was intraperitoneally injected in model group.After 45 min of ischemia and 6 h of reperfusion, rats were sacrificed.Serum and ileum were collected for further detection.Tumor necrosis factor alpha (TNF-α), interleukin (IL)- 1β and IL-17A expression levels in serum were detected by conducting enzyme-linked immunosorbent assay (ELISA). The localization expressions of tight junction protein Occludin was detected by immunohistochemical staining (IHC), while the localization expression of tight junction protein zonula occluden-1 (ZO-1) were detected by immunofluorescence staining (IF). The protein expression of Occludin, ZO-1, and Lysozyme were detected by performing Western blot.The mRNA expression of Lysozyme and α-defensin were detected by real-time PCR (qPCR).Results:ELISA results proved that the levels of serum TNF-α and IL-1β in HRS group rats were significantly lower than those in model group [(62.02±29.97) ng/Ln vs.(113.40±44.58) ng/L, (21.68±0.35) ng/L n vs.(28.29±3.49) ng/L], while the level of IL-17A increased [(28.18±5.28) ng/L n vs. (15.10±3.60) ng/L] (all n P<0.05). IHC staining: compared with model group, the expression of Occludin in HRS group was uniform and continuous, and the staining was darker.IF results: compared with model group, the fluorescence signal intensity of ZO-1 in HRS group rats significantly increased, and the distribution was clear and continuous.Wes-tern blot results: compared with model group, the expression levels of Occludin and ZO-1 proteins in HRS group rats remarkably increased (0.79±0.06n vs. 0.54±0.04, 0.91±0.11 n vs. 0.51±0.13), while Lysozyme protein decreased (1.50±0.40 n vs. 2.99±0.80) (all n P<0.05). qPCR results revealed that the expression level of Lysozyme mRNA in HRS group rats was lower than that in model group (1.64±0.33n vs. 2.20±0.40), while α-defensin mRNA obviously increased (0.82±0.19 n vs. 0.47±0.13) (all n P<0.01).n Conclusions:HRS protects intestinal mucosal barrier by inhibiting the expression of tight junctions and the secretion of antimicrobial peptides in rat suffering from IIRI.