甲烷盐水在脓毒症大鼠肠道损伤中的保护作用
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摘要
目的观察甲烷盐水对脓毒症大鼠肠道损伤的保护作用,并探讨其可能的作用机制。方法雄性SD大鼠128只,体重220～280 g,8～10周龄,采用随机数字表法,分为4组(n=32):假手术组(Sham组)、脓毒症组(Sepsis组)、甲烷盐水组(Sham+MS组)和脓毒症甲烷盐水组(Sepsis+MS组)。采用盲肠结扎穿孔(CLP)法制备脓毒症模型。CLP术后0.5 h和12 h时Sham+MS组和Sepsis+MS组腹腔注入10%甲烷盐水10 mL/kg。每组取20只大鼠,观察术后7 d内的生存率。CLP术后6、18 h处死大鼠,取中段空肠染色并作组织病理学Chiu′S评分;检测各组大鼠肠组织超氧化物歧化酶(SOD)、髓组织过氧化物酶含量(MPO)、肿瘤坏死因子-α(TNF-α)和白细胞介素-10(IL-10)水平,采用Western blot测肠道组织核因子E2相关因子2(Nrf2)、血红素氧合酶-1(HO-1)和肠道组织紧密连接蛋白(Occludin)的蛋白表达。结果与Sham组比较,Sepsis组和Sepsis+MS组肠道组织病理学Chiu′S评分升高,Nrf2、HO-1和Occludin蛋白表达上调(P<0.05),Sham+MS组上述各指标差异无统计学意义(P>0.05);与Sepsis组比较,Sepsis+MS组肠道组织病理学Chiu′S评分降低,Nrf2、HO-1和Occludin蛋白表达上调,差异有统计学意义(P<0.05)。结论甲烷盐水治疗减轻脓毒症大鼠肠道组织损伤的机制与激活Nrf2/ARE通路上调HO-1促进肠道Occludin表达有关。
Objective To investigate the role of HO-1 on methane saline treatment for intestinal injury caused by sepsis and to explore its possible mechanism. Methods A total of 128 male SD rats(weighing 220-260 g and 8-10 weeks old) were randomly divided into 4 groups(n=32), Group Sham, Group Sepsis, Group Sham+MS and Group Sepsis+MS. Sepsis model was established by cecal ligation and puncture(CLP). Half hour and 12 h after CLP, rats in Group Sham+MS and Sepsis+MS were intraperitoneally injected with 10% methane saline(10 mL/kg). Twenty rats in each group were used to observe the survival rate within 7 days after operation. CLP rats were sacrificed 6 h and 18 h after operation, and the jejunum tissues were collected for evaluation of histolopathological Chiu′s score and intestinal permeability. The peroxidation of superoxide dismutase(SOD), myeloperoxidase(MPO), tumor necrosis factor alpha(TNF-α) and interleukin-10(IL-10) in the intestinal mucosa were detected by Elisa. The expression of Nrf2, HO-1 and tight junction protein(Occludin) were assessed by Western blot.Results Compared with Group Sham, the Chiu′s scores were significantly increased, while the expression of Nrf2, HO-1 and Occludin proteins were significantly up-regulated in Group Sepsis and Sepsis+MS(P<0.05). Compared with Group Sepsis, Chiu′s score in Group Sepsis+MS was significantly decreased, but the expression of Nrf2, HO-1 and Occludin proteins were significantly up-regulated(P<0.05).Conclusion The mechanism of methane saline in reducing intestinal tissue injury in rats with sepsis is correlated to the activation of Nrf2/ARE pathway and the up-regulation of HO-1, thus to promote intestinal Occludin expression.
Objective To investigate the role of HO-1 on methane saline treatment for intestinal injury caused by sepsis and to explore its possible mechanism. Methods A total of 128 male SD rats(weighing 220-260 g and 8-10 weeks old) were randomly divided into 4 groups(n=32), Group Sham, Group Sepsis, Group Sham+MS and Group Sepsis+MS. Sepsis model was established by cecal ligation and puncture(CLP). Half hour and 12 h after CLP, rats in Group Sham+MS and Sepsis+MS were intraperitoneally injected with 10% methane saline(10 mL/kg). Twenty rats in each group were used to observe the survival rate within 7 days after operation. CLP rats were sacrificed 6 h and 18 h after operation, and the jejunum tissues were collected for evaluation of histolopathological Chiu′s score and intestinal permeability. The peroxidation of superoxide dismutase(SOD), myeloperoxidase(MPO), tumor necrosis factor alpha(TNF-α) and interleukin-10(IL-10) in the intestinal mucosa were detected by Elisa. The expression of Nrf2, HO-1 and tight junction protein(Occludin) were assessed by Western blot.Results Compared with Group Sham, the Chiu′s scores were significantly increased, while the expression of Nrf2, HO-1 and Occludin proteins were significantly up-regulated in Group Sepsis and Sepsis+MS(P<0.05). Compared with Group Sepsis, Chiu′s score in Group Sepsis+MS was significantly decreased, but the expression of Nrf2, HO-1 and Occludin proteins were significantly up-regulated(P<0.05).Conclusion The mechanism of methane saline in reducing intestinal tissue injury in rats with sepsis is correlated to the activation of Nrf2/ARE pathway and the up-regulation of HO-1, thus to promote intestinal Occludin expression.
引文
[1] Hotchkiss RS, Karl IE. The pathophysiology and treatment of sepsis[J]. New Engl J Med, 2003,348(2): 138-150.
[2] 李媛, 谢克亮, 陈红光,等. Nrf2在氢气治疗严重脓毒症肠损伤中的作用[J]. 中华危重病急救医学, 2014, 26(6):415-419.
[3] Chen Z, Zhang Y, Liang M, et al. Nrf2 plays a pivotal role in protection against burn trauma-induced intestinal injury and death[J]. Oncotarget, 2016,7(15): 19272-19283.
[4] Li Y, Xie K, Chen H, et al. Hydrogen gas inhibits high-mobility group box 1 release in septic mice by upregulation of heme oxygenase 1[J]. J Surg Res, 2015,196(1): 136-148.
[5] Kensler TW, Wakabayashi N, Biswal S. Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway[J]. Annu Rev Pharmacol Toxico, 2007,47(1): 89-116.
[6] Nguyen T, Nioi P, Pickett CB. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress[J]. J Bio Chem, 2009,284(20): 13291-13295.
[7] Chai J, Luo L, Hou F, et al. Agmatine Reduces Lipopolysaccharide-Mediated Oxidant Response via Activating PI3K/Akt Pathway and Up-Regulating Nrf2 and HO-1 Expression in Macrophages[J]. Plos One, 2016,11(9): e0163634.
[8] 陈广响, 任乐豪, 单可树, 等. 缺氧条件下血红素氧合酶-1对肠黏膜屏障的保护作用[J]. 中华实验外科杂志, 2014,31(10): 2169-2171.
[9] Chen O, Ye Z, Cao Z, et al. Methane attenuates myocardial ischemia injury in rats through anti-oxidative, anti-apoptotic and anti-inflammatory actions[J]. Free Radic Biol Med, 2015,90: 1-11.
[10] Xu Z, Na L, Han S, et al. Methane limit LPS-induced NF-κB/MAPKs signal in macrophages and suppress immune response in mice by enhancing PI3K/AKT/GSK-3β-mediated IL-10 expression[J]. Sci Rep-UK, 2016,6: 29359.
[11] Wang L, Yao Y, He R, et al. Methane ameliorates spinal cord ischemia-reperfusion injury in rats: Antioxidant, anti-inflammatory and anti-apoptotic activity mediated by Nrf2 activation[J]. Free Radic Biol Med, 2017,103: 69-86.
[12] Wu JY, Tsou MY, Chen TH, et al. Therapeutic effects of melatonin on peritonitis-induced septic shock with multiple organ dysfunction syndrome in rats[J]. J Pineal Res, 2008,45(1): 106-116.
[13] Chiu CJ, Mcardle AH, Brown R, et al. Intestinal Mucosal Lesion in Low-Flow States: I. A Morphological, Hemodynamic, and Metabolic Reappraisal[J]. Arch Surg- Chicago, 1970,101(4): 478-483.
[14] Piliponsky AM, Chen CC, Nishimura T, et al. Neurotensin increases mortality and mast cells reduce neurotensin levels in a mouse model of sepsis[J]. Nat Med, 2008,14(4): 392-398.
[15] 朱雪琦, 刘清泉, 姚咏明. 脓毒症动物模型制备方法的研究进展[J]. 中华危重病急救医学, 2006,18(2): 114-116.
[16] 韩磊, 任爱民. 脓毒症胃肠功能损伤机制及保护修复的研究进展[J]. 中国中西医结合急救杂志,2009,16(5): 314-317.
[17] Yuhai GU, Zhao Z. Significance of the changes occurring in the levels of interleukins, SOD and MDA in rat pulmonary tissue following exposure to different altitudes and exposure times[J]. Exp Ther Med, 2015,10(3): 915-920.
[18] Nakazato T, Sagawa M, Yamato K, et al. Myeloperoxidase is a key regulator of oxidative stress mediated apoptosis in myeloid leukemic cells[J]. J Am Assoc Cancer Res, 2007,13(18 Pt 1): 5436-5445.
[19] Rocha-Ferreira E, Kelen D, Faulkner S, et al. Systemic pro-inflammatory cytokine status following therapeutic hypothermia in a piglet hypoxia-ischemia model[J]. J Neuroinflamm, 2017,14(1): 44.
[20] Li X, Liu X, Yu S. Psychological Stress-Derived Prolactin Modulates Occludin Expression in Vaginal Epithelial Cells to Compromise Barrier Function[J]. Cell Physiol Biochem, 2015,37(1): 153-161.
[21] Ma Q. Role of Nrf2 in Oxidative Stress and Toxicity[J]. Annu Rev Pharmacol Toxicol, 2013,53(1): 401-426.
[22] 朱子夫. HO-1抗氧化损伤的研究进展[J]. 医学综述, 2010,16(15): 2266-2270.
[23] Tan Y, Wang Q, She Y, et al. Ketamine reduces LPS-induced HMGB1 via activation of the Nrf2/HO-1 pathway and NF-κB suppression[J]. J Tr Acute Care Surg, 2015,78(4): 784-792.
[24] 骆德强, 陈自力. 甲烷的抗炎作用及其机制研究进展[J]. 中国中西医结合急救杂志, 2017,24(1): 107-110.
[25] Zhang L, Liu YL, Chen GX, et al. Heme oxygenase-1 promotes Caco-2 cell proliferation and migration by targeting CTNND1[J]. Chin J Ophthalmol, 2013,126(16): 3057-3063.
[2] 李媛, 谢克亮, 陈红光,等. Nrf2在氢气治疗严重脓毒症肠损伤中的作用[J]. 中华危重病急救医学, 2014, 26(6):415-419.
[3] Chen Z, Zhang Y, Liang M, et al. Nrf2 plays a pivotal role in protection against burn trauma-induced intestinal injury and death[J]. Oncotarget, 2016,7(15): 19272-19283.
[4] Li Y, Xie K, Chen H, et al. Hydrogen gas inhibits high-mobility group box 1 release in septic mice by upregulation of heme oxygenase 1[J]. J Surg Res, 2015,196(1): 136-148.
[5] Kensler TW, Wakabayashi N, Biswal S. Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway[J]. Annu Rev Pharmacol Toxico, 2007,47(1): 89-116.
[6] Nguyen T, Nioi P, Pickett CB. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress[J]. J Bio Chem, 2009,284(20): 13291-13295.
[7] Chai J, Luo L, Hou F, et al. Agmatine Reduces Lipopolysaccharide-Mediated Oxidant Response via Activating PI3K/Akt Pathway and Up-Regulating Nrf2 and HO-1 Expression in Macrophages[J]. Plos One, 2016,11(9): e0163634.
[8] 陈广响, 任乐豪, 单可树, 等. 缺氧条件下血红素氧合酶-1对肠黏膜屏障的保护作用[J]. 中华实验外科杂志, 2014,31(10): 2169-2171.
[9] Chen O, Ye Z, Cao Z, et al. Methane attenuates myocardial ischemia injury in rats through anti-oxidative, anti-apoptotic and anti-inflammatory actions[J]. Free Radic Biol Med, 2015,90: 1-11.
[10] Xu Z, Na L, Han S, et al. Methane limit LPS-induced NF-κB/MAPKs signal in macrophages and suppress immune response in mice by enhancing PI3K/AKT/GSK-3β-mediated IL-10 expression[J]. Sci Rep-UK, 2016,6: 29359.
[11] Wang L, Yao Y, He R, et al. Methane ameliorates spinal cord ischemia-reperfusion injury in rats: Antioxidant, anti-inflammatory and anti-apoptotic activity mediated by Nrf2 activation[J]. Free Radic Biol Med, 2017,103: 69-86.
[12] Wu JY, Tsou MY, Chen TH, et al. Therapeutic effects of melatonin on peritonitis-induced septic shock with multiple organ dysfunction syndrome in rats[J]. J Pineal Res, 2008,45(1): 106-116.
[13] Chiu CJ, Mcardle AH, Brown R, et al. Intestinal Mucosal Lesion in Low-Flow States: I. A Morphological, Hemodynamic, and Metabolic Reappraisal[J]. Arch Surg- Chicago, 1970,101(4): 478-483.
[14] Piliponsky AM, Chen CC, Nishimura T, et al. Neurotensin increases mortality and mast cells reduce neurotensin levels in a mouse model of sepsis[J]. Nat Med, 2008,14(4): 392-398.
[15] 朱雪琦, 刘清泉, 姚咏明. 脓毒症动物模型制备方法的研究进展[J]. 中华危重病急救医学, 2006,18(2): 114-116.
[16] 韩磊, 任爱民. 脓毒症胃肠功能损伤机制及保护修复的研究进展[J]. 中国中西医结合急救杂志,2009,16(5): 314-317.
[17] Yuhai GU, Zhao Z. Significance of the changes occurring in the levels of interleukins, SOD and MDA in rat pulmonary tissue following exposure to different altitudes and exposure times[J]. Exp Ther Med, 2015,10(3): 915-920.
[18] Nakazato T, Sagawa M, Yamato K, et al. Myeloperoxidase is a key regulator of oxidative stress mediated apoptosis in myeloid leukemic cells[J]. J Am Assoc Cancer Res, 2007,13(18 Pt 1): 5436-5445.
[19] Rocha-Ferreira E, Kelen D, Faulkner S, et al. Systemic pro-inflammatory cytokine status following therapeutic hypothermia in a piglet hypoxia-ischemia model[J]. J Neuroinflamm, 2017,14(1): 44.
[20] Li X, Liu X, Yu S. Psychological Stress-Derived Prolactin Modulates Occludin Expression in Vaginal Epithelial Cells to Compromise Barrier Function[J]. Cell Physiol Biochem, 2015,37(1): 153-161.
[21] Ma Q. Role of Nrf2 in Oxidative Stress and Toxicity[J]. Annu Rev Pharmacol Toxicol, 2013,53(1): 401-426.
[22] 朱子夫. HO-1抗氧化损伤的研究进展[J]. 医学综述, 2010,16(15): 2266-2270.
[23] Tan Y, Wang Q, She Y, et al. Ketamine reduces LPS-induced HMGB1 via activation of the Nrf2/HO-1 pathway and NF-κB suppression[J]. J Tr Acute Care Surg, 2015,78(4): 784-792.
[24] 骆德强, 陈自力. 甲烷的抗炎作用及其机制研究进展[J]. 中国中西医结合急救杂志, 2017,24(1): 107-110.
[25] Zhang L, Liu YL, Chen GX, et al. Heme oxygenase-1 promotes Caco-2 cell proliferation and migration by targeting CTNND1[J]. Chin J Ophthalmol, 2013,126(16): 3057-3063.