虾青素对小鼠结肠急性氧化损伤的保护作用
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摘要
试验旨在研究虾青素对脂多糖(LPS)诱导的小鼠急性结肠炎及氧化应激的影响。选择6周龄健康雄性ICR小鼠32只,随机分为4组,其中对照组和LPS组小鼠连续15 d灌胃橄榄油,虾青素组和虾青素保护组小鼠连续15 d灌胃50 mg/kg虾青素,15 d后对照组和虾青素组腹腔注射生理盐水,LPS组和虾青素保护组腹腔注射1 mg/kg LPS,3 h后处死,采集血液及结肠组织。ELISA和荧光定量PCR检测血清及结肠中炎性因子和抗氧化酶水平,生物化学法测定结肠中氧化水平,Western blot检测结肠中p-NF-κB p65蛋白相对表达量,HE染色观察结肠病理形态学变化。结果表明:与对照组相比,LPS组小鼠血清和结肠组织中炎症因子的表达及NF-κB p65蛋白的活化均显著升高(P<0.05),组织抗氧化水平低(P<0.05),结肠黏膜损伤严重;与LPS组相比,保护组小鼠血清和结肠组织中炎症因子表达及NF-κB p65蛋白的活化均显著降低(P<0.05),组织抗氧化水平高(P<0.05),且结肠黏膜损伤程度低。虾青素可保护小鼠结肠黏膜结构,通过抑制NF-κB信号通路降低机体中炎性因子的分泌,提高组织抗氧化水平,缓解LPS引起的急性结肠损伤。
To investigate the effects of astaxanthin(ATX) on acute colon inflammation and oxidative stress induced by lipopolysaccharide(LPS) in mice. Thirty-two healthy male ICR mice were randomly allocated into four groups. ATX and olive oil was administrated by oral gavage for 15 d consecutively before the saline or LPS(1 mg/kg) injected. Blood and colon tissues were collected after 3 h later. The levels of inflammatory factors and antioxidant enzyme in serum or colon were measured by ELISA or fluorescence quantitative PCR. The activities of antioxidant enzyme were measured by biochemical methods. p-NF-κB p65 protein was measured by western blot. H&E staining was used to observe the histopathological changes. Compared with the control group, the expression of inflammatory factors and the activation of NF-κB p65 protein in serum and colon tissues of LPS group were significantly increased(P<0.05), the tissue antioxidant level was low, and the colonic mucosa was severely damaged. Compared with LPS group, the expression of inflammatory factors and the activation of NF-κB p65 protein in serum and colon tissues of the protective group were significantly decreased(P<0.05), and the tissue antioxidant level was high. In addition, the degree of colon mucosal damage was low. ATX treatment can protect colonic mucosa structure in mice, and relieve acute colon injury by suppressing the secretion of inflammatory cytokines, increase antioxidant levels and inhibiting the activation of NF-κB p65 protein in NF-κB pathway in LPS-induced mice.
To investigate the effects of astaxanthin(ATX) on acute colon inflammation and oxidative stress induced by lipopolysaccharide(LPS) in mice. Thirty-two healthy male ICR mice were randomly allocated into four groups. ATX and olive oil was administrated by oral gavage for 15 d consecutively before the saline or LPS(1 mg/kg) injected. Blood and colon tissues were collected after 3 h later. The levels of inflammatory factors and antioxidant enzyme in serum or colon were measured by ELISA or fluorescence quantitative PCR. The activities of antioxidant enzyme were measured by biochemical methods. p-NF-κB p65 protein was measured by western blot. H&E staining was used to observe the histopathological changes. Compared with the control group, the expression of inflammatory factors and the activation of NF-κB p65 protein in serum and colon tissues of LPS group were significantly increased(P<0.05), the tissue antioxidant level was low, and the colonic mucosa was severely damaged. Compared with LPS group, the expression of inflammatory factors and the activation of NF-κB p65 protein in serum and colon tissues of the protective group were significantly decreased(P<0.05), and the tissue antioxidant level was high. In addition, the degree of colon mucosal damage was low. ATX treatment can protect colonic mucosa structure in mice, and relieve acute colon injury by suppressing the secretion of inflammatory cytokines, increase antioxidant levels and inhibiting the activation of NF-κB p65 protein in NF-κB pathway in LPS-induced mice.
引文
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[6]Reuter S,Gupta S C,Chaturvedi M M,et al.Oxidative stress,inflammation,and cancer:how are they linked?[J].Free Radical Bio Med,2010,49(11):1603-1616.
[7]Pahl H L.Activators and target genes of Rel/NF-κB transcription factors[J].Oncogene,1999,18(49):6853.
[8]Balkwill F,Mantovani A.Inflammation and cancer:back to Virchow?[J].Lancet,2001,357(9255):539-545.
[9]Nagendraprabhu P,Sudhandiran G.Astaxanthin inhibits tumor invasion by decreasing extracellular matrix production and induces apoptosis in experimental rat colon carcinogenesis by modulating the expressions of ERK-2,NFkB and COX-2[J].Invest New Drug,2011,29(2):207-224.
[10]Tanwar L,Vaish V,Sanyal S N.Chemoprevention of 1,2-dimethylhydrazine-induced colon carcinogenesis by a non-steroidal anti-inflammatory drug,etoricoxib,in rats:Inhibition of nuclear factor kappaB[J].Asian Pac J Cancer Prev,2009,10:1141-1146.
[11]Qin X,Qiu C,Zhao L.Lysophosphatidylcholine perpetuates macrophage polarization toward classically activated phenotype in inflammation[J].Cell Immunol,2014,289(1-2):185-190.
[12]Johnson E R,Matthay M A.Acute lung injury:epidemiology,pathogenesis,and treatment[J].J Aerosol Med Pulm D,2010,23(4):243-252.
[13]Zhou L,Gao M,Xiao Z,et al.Protective effect of astaxanthin against multiple organ injury in a rat model of sepsis[J].J Surg Res,2015,195(2):559-567.
[14]Bi J,Cui R,Li Z,et al.Astaxanthin alleviated acute lung injury by inhibiting oxidative/nitrative stress and the inflammatory response in mice[J].Biomed Pharmacother,2017,95:974-982.
[15]Cao J,Wang W.Effects of astaxanthin and esterified glucomannan on hematological and serum parameters,and liver pathological changes in broilers fed aflatoxin-B1-contaminated feed[J].Anim Sci J,2014,85(2):150-157.
[16]Abraham E,Carmody A,Shenkar R,et al.Neutrophils as early immunologic effectors in hemorrhage-or endotoxemia-induced acute lung injury[J].Am J Physiol Lung Cell Mol Physiol,2000,279(6):L1137-L1145.
[17]Cicho?-Lach H,Michalak A.Oxidative stress as a crucial factor in liver diseases[J].World J Gastroenterol,2014,20(25):8082-8091.
[18]Urso M L,Clarkson P M.Oxidative stress,exercise,and antioxidant supplementation[J].Toxicology,2003,189(1-2):41-54.
[19]Aktan F.iNOS-mediated nitric oxide production and its regulation[J].Life Sci,2004,75(6):639-653.
[20]Al-Amin M M,Reza H M,Saadi H M,et al.Astaxanthin ameliorates aluminum chloride-induced spatial memory impairment and neuronal oxidative stress in mice[J].Eur J Pharmacol,2016,777:60-69.
[21]Park J S,Chyun J H,Kim Y K,et al.Astaxanthin decreased oxidative stress and inflammation and enhanced immune response in humans[J].Nutr Metab,2010,7(1):18.