烟雾所致轻度稳定期COPD小鼠肺组织中MRP1表达增加与Nrf2信号通路有关
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摘要
目的探讨Nrf2信号通路对烟雾致轻度稳定期COPD小鼠肺组织中MRP1表达的影响。方法被动香烟烟吸法建立轻度COPD小鼠模型,检测肺功能;病理切片观察肺组织病理学改变;免疫组化及Western blot检测相关蛋白在肺组织中的表达水平。结果与正常组相比,野生型(WT)及Nrf2~(-/-)模型组各肺功能指标明显降低;并且Nrf2~(-/-)模型组与WT模型组相比,各肺功能指标的下降更为明显。HE染色结果显示,WT及Nrf2~(-/-)模型小鼠肺泡中均发生弥漫性炎症反应,肺泡支气管结构受损,并且在Nrf2~(-/-)模型小鼠中病理改变更为明显。免疫组化及Western blot结果显示,与WT正常组相比,MRP1在Nrf2~(-/-)正常组小鼠肺组织中的表达明显减少;被动香烟烟吸后,与WT正常组相比,MRP1、Nrf2、HO-1在WT模型组中的表达明显增多,但是与Nrf2~(-/-)正常组相比,在Nrf2~(-/-)模型组中MRP1的表达并未发生明显改变。结论 香烟烟雾所致轻度稳定期COPD小鼠肺组
Aim To investigate the effect of Nrf2 pathway on the expression of MRP1 in mildly stable COPD mice. Methods The mild COPD mouse model was established by passive cigarette smoking. The pathological changes of lung tissues were examined by HE staining. Immunohistochemistry and Western blot were used to detect the protein expression of MRP1, Nrf2 and HO-1.Results Compared with normal group, each lung function index of the mild-moderate COPD model group was significantly lower, but compared with wide type(WT) model group, the reduction was more significant in Nrf2~(-/-) model group. HE results showed diffuse inflammatory reaction and alveolar bronchial structure damage in alveolar of WT and Nrf2~(-/-) model mice, and it was more pronounced in Nrf2~(-/-) mice. Immunohistochemistry and Western blot results showed that the expression of MRP1 in lung tissue of Nrf2~(-/-) normal mice was significantly reduced compared with the normal WT group. After passive cigarette smoking, The expression of MRP1, Nrf2 and HO-1 in WT model group increased significantly, but compared with Nrf2~(-/-) normal mice, there was no significant change in the expression of MRP1 in Nrf2~(-/-) model group.Conclusions Mildly stable COPD mice may counteract the xenobiotic damage caused by cigarette smoke through up-regulating the expression of MRP1 protein, which may be associated with Nrf2 signaling activation.
Aim To investigate the effect of Nrf2 pathway on the expression of MRP1 in mildly stable COPD mice. Methods The mild COPD mouse model was established by passive cigarette smoking. The pathological changes of lung tissues were examined by HE staining. Immunohistochemistry and Western blot were used to detect the protein expression of MRP1, Nrf2 and HO-1.Results Compared with normal group, each lung function index of the mild-moderate COPD model group was significantly lower, but compared with wide type(WT) model group, the reduction was more significant in Nrf2~(-/-) model group. HE results showed diffuse inflammatory reaction and alveolar bronchial structure damage in alveolar of WT and Nrf2~(-/-) model mice, and it was more pronounced in Nrf2~(-/-) mice. Immunohistochemistry and Western blot results showed that the expression of MRP1 in lung tissue of Nrf2~(-/-) normal mice was significantly reduced compared with the normal WT group. After passive cigarette smoking, The expression of MRP1, Nrf2 and HO-1 in WT model group increased significantly, but compared with Nrf2~(-/-) normal mice, there was no significant change in the expression of MRP1 in Nrf2~(-/-) model group.Conclusions Mildly stable COPD mice may counteract the xenobiotic damage caused by cigarette smoke through up-regulating the expression of MRP1 protein, which may be associated with Nrf2 signaling activation.
引文
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[6] Fratta Pasini A M, Ferrari M, Stranieri C, et al. Nrf2 expression is increased in peripheral blood mononuclear cells derived from mild-moderate ex-smoker COPD patients with persistent oxidative stress[J]. Int J Chron Obstruct Pulmon Dis, 2016, 11: 1733-43.
[7] 曹君, 陈平, 杨悦, 等. 烟雾暴露所致肺气肿小鼠模型的建立与评价[J]. 中国实验动物学报, 2010, 18(4): 278-83. Cao J, Chen P, Yang Y, et al. Establishment and evaluation of mouse models of emphysema induced by smoke exposure[J]. Chin J Exp Anim, 2010,18(4): 278-83.
[8] 高振, 哈木拉提·吾甫尔, 廖春燕. 慢性阻塞性肺疾病动物模型研究概况[J]. 国际病理科学与临床杂志, 2012, 32(2): 169-74. Gao Z, Hammurati.Ugur, Liao C Y. A survey of animal models of chronic obstructive pulmonary disease[J]. Int J Pathol Sci Clin Med, 2012, 32(2): 169-74.
[9] 张兰英, 张婧, 欧阳瑶. 烟熏诱导慢性阻塞性肺疾病小鼠模型的建立及验证[J]. 山东医药, 2016, 56(32): 35-8. Zhang L Y, Zhang J, OuYang Y. Flue-induced chronic obstructive pulmonary disease mouse model establishment and validation[J]. Shandong Med, 2016, 56(32): 35-8.
[10] 赵萍, 高建. Nrf2/Keap1/ARE信号通路与难治性呼吸系统疾病研究进展[J].中国药理学通报, 2015, 31(11): 1488-92. Zhao P, Gao J. Nrf2/Keap1/ARE signaling pathway and refractory respiratory disease research progress[J]. Chin Pharmacol Bull, 2015, 31(11): 1488-92.
[11] 田戈, 姜敏. Nrf2-ARE信号通路在调节慢性阻塞性肺疾病氧化失衡过程中的作用机制[J]. 广东医学, 2016, 37(12): 1808-10. Tian G, Jiang M. Role of Nrf2-ARE signaling pathway in regulation of oxidative imbalance in chronic obstructive pulmonary diseases[J]. Guangdong Med, 2016, 37(12): 1808-10.
[12] Paumi C M, Chuk M, Snider J, et al. ABC transporters in Saccharomyces cerevisiae and their interactors: new technology advances the biology of the ABCC(MRP) subfamily[J]. Microbiol Mol Biol Rev, 2009, 73(4): 577-93.
[13] Xu X, Zhang Y, Li W, et al. Wogonin reverses multi-drug resistance of human myelogenous leukemia K562/A02 cells via downregulation of MRP1 expression by inhibiting Nrf2/ARE signaling pathway[J]. Biochem Pharmacol, 2014, 92(2): 220-34.
[14] Maher J M, Dieter M Z, Aleksunes L M, et al. Oxidative and electrophilic stress induces multidrug resistance-associated protein transporters via the nuclear factor-E2-related factor-2 transcriptional pathway[J]. Hepatology,2007,46(5): 1597-610.
[15] Perez M J, Gonzalez-Sanchez E, Gonzalea-Loyola A, et al. Mitochondrial genome depletion dysregulates bile acid- and paracetamol-induced expression of the transporters Mdr1, Mrp1, Mrp4 in liver cells[J]. Br J Pharmacol, 2011, 162(8): 1686-99.
[2] Budulac S E, Postma D S, Hiemstra P S, et al. Multidrug resistance-associated protein-1(MRP1) genetic variants, MRP1 protein levels and severity of COPD[J]. Respir Res, 2010, 11: 60.
[3] 李航, 段惠军. Nrf2/ARE信号通路及其调控的抗氧化蛋白[J]. 中国药理学通报, 2011, 27(3) : 300-3. Li H, Duan H J. Nrf2/ARE signaling pathway and its regulatory antioxidant proteins[J].Chin Pharmacol Bull, 2011, 27(3) : 300-3.
[4] Van der Deen M, Marks H, Willemse B W, et al. Diminished expression of multidrug resistance-associated protein 1(MRP1) in bronchial epithelium of COPD patients[J]. Virchows Arch, 2006, 449(6): 682-8.
[5] Malhotra D, Thimmulappa R, Navas-Acien A, et al. Decline in NRF2-regulated antioxidants in chronic obstructive pulmonary disease lungs due to loss of its positive regulator, DJ-1[J]. Am J Respir Crit Care Med, 2008, 178(6): 592-604.
[6] Fratta Pasini A M, Ferrari M, Stranieri C, et al. Nrf2 expression is increased in peripheral blood mononuclear cells derived from mild-moderate ex-smoker COPD patients with persistent oxidative stress[J]. Int J Chron Obstruct Pulmon Dis, 2016, 11: 1733-43.
[7] 曹君, 陈平, 杨悦, 等. 烟雾暴露所致肺气肿小鼠模型的建立与评价[J]. 中国实验动物学报, 2010, 18(4): 278-83. Cao J, Chen P, Yang Y, et al. Establishment and evaluation of mouse models of emphysema induced by smoke exposure[J]. Chin J Exp Anim, 2010,18(4): 278-83.
[8] 高振, 哈木拉提·吾甫尔, 廖春燕. 慢性阻塞性肺疾病动物模型研究概况[J]. 国际病理科学与临床杂志, 2012, 32(2): 169-74. Gao Z, Hammurati.Ugur, Liao C Y. A survey of animal models of chronic obstructive pulmonary disease[J]. Int J Pathol Sci Clin Med, 2012, 32(2): 169-74.
[9] 张兰英, 张婧, 欧阳瑶. 烟熏诱导慢性阻塞性肺疾病小鼠模型的建立及验证[J]. 山东医药, 2016, 56(32): 35-8. Zhang L Y, Zhang J, OuYang Y. Flue-induced chronic obstructive pulmonary disease mouse model establishment and validation[J]. Shandong Med, 2016, 56(32): 35-8.
[10] 赵萍, 高建. Nrf2/Keap1/ARE信号通路与难治性呼吸系统疾病研究进展[J].中国药理学通报, 2015, 31(11): 1488-92. Zhao P, Gao J. Nrf2/Keap1/ARE signaling pathway and refractory respiratory disease research progress[J]. Chin Pharmacol Bull, 2015, 31(11): 1488-92.
[11] 田戈, 姜敏. Nrf2-ARE信号通路在调节慢性阻塞性肺疾病氧化失衡过程中的作用机制[J]. 广东医学, 2016, 37(12): 1808-10. Tian G, Jiang M. Role of Nrf2-ARE signaling pathway in regulation of oxidative imbalance in chronic obstructive pulmonary diseases[J]. Guangdong Med, 2016, 37(12): 1808-10.
[12] Paumi C M, Chuk M, Snider J, et al. ABC transporters in Saccharomyces cerevisiae and their interactors: new technology advances the biology of the ABCC(MRP) subfamily[J]. Microbiol Mol Biol Rev, 2009, 73(4): 577-93.
[13] Xu X, Zhang Y, Li W, et al. Wogonin reverses multi-drug resistance of human myelogenous leukemia K562/A02 cells via downregulation of MRP1 expression by inhibiting Nrf2/ARE signaling pathway[J]. Biochem Pharmacol, 2014, 92(2): 220-34.
[14] Maher J M, Dieter M Z, Aleksunes L M, et al. Oxidative and electrophilic stress induces multidrug resistance-associated protein transporters via the nuclear factor-E2-related factor-2 transcriptional pathway[J]. Hepatology,2007,46(5): 1597-610.
[15] Perez M J, Gonzalez-Sanchez E, Gonzalea-Loyola A, et al. Mitochondrial genome depletion dysregulates bile acid- and paracetamol-induced expression of the transporters Mdr1, Mrp1, Mrp4 in liver cells[J]. Br J Pharmacol, 2011, 162(8): 1686-99.
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