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[摘要] 目的 探討黄精多糖（PSP）体外抗氧化作用及其对小鼠炎症性肠病是否具有保护作用。 方法 体外抗氧化实验观察PSP对羟自由基（·OH）的产生、·OH对红细胞破坏作用及肝匀浆脂质产生是否具有抑制作用；建立小鼠炎症性肠病模型并给予PSP治疗，观察小鼠疾病活动度积分（DAI）评分，检测小鼠结肠组织中MDA、SOD、MPO，并应用Western blot对小鼠结肠组织中NF-κB蛋白表达进行测定。 结果 PSP在体外对·OH的产生及·OH对红细胞破坏具有抑制作用（P<0.01）、抑制肝匀浆脂质过氧化过程（P<0.01），且其体外抗氧化作用具有剂量依赖性（P<0.05～0.01）；与模型组相比，PSP能降低DAI评分（P<0.01）、抑制结肠组织中MDA、MPO产生（P<0.05～0.01），增加SOD含量（P<0.05），并对NF-κB蛋白表达具有抑制作用（P<0.05）。 结论 黄精多糖具有体外抗氧化作用，其对小鼠炎症性肠病的抑制作用可能是抑制NF-κB通路，通过减少MDA、MPO产生和增加SOD含量实现的。
[关键词] 黄精多糖；炎症性肠病；体外抗氧化能力；NF-κB通路
[中图分类号] R285 [文献标识码] A [文章编号] 1673-9701（2017）29-0027-04
Study on the antioxidant effect in vitro of polygonatum sibiricum polysaccharides and its effect on inflammatory bowel disease in mice
XUE Xuebin1 FANG Shuhua2 WANG Huajun3
1.Department of Drug and Equipment， Zhenjiang Second People’s Hospital in Jiangsu Province， Zhenjiang 212000， China； 2.Department of Pharmacy， Zhenjiang First People’s Hospital in Jiangsu Province， Zhenjiang 212000， China； 3.Department of Pharmacy， the Affiliated Hospital of Jiangsu University， Zhenjiang 212001， China
[Abstract] Objective To investigate the anti-oxidative effects of polygonatum sibiricum polysaccharides（PSP） in vitro and its protective effect on inflammatory bowel disease in mice. Methods In vitro anti-oxidative experiment was carried out to observe the effect of PSP on the generation of hydroxyl radical（·OH）， the effect of ·OH on the destruction of erythrocytes and whether lipid production of liver homogenate had inhibitory effect. The model of mice inflammatory bowel disease was established and was given PSP treatment. Mice disease activity score（DAI） was calculated， MDA， SOD and MPO in colonic tissue of mice were detected， and the expression of NF-κB protein in the colonic tissue of mice was determined. Results PSP showed inhibitory effect on the generation of ·OH in vitro and ·OH showed inhibitory effect on the destruction of erythrocytes（P<0.01）， which inhibited liver homogenate lipid peroxidation process（P<0.01）， and it’s in vitro anti-oxidant effect was dose-dependent（P<0.05-0.01）. Compared with the model group， PSP can reduce the DAI score（P<0.01）， inhibit the generation of MDA and MPO in colonic tissues（P<0.05-0.01）， and increase the content of SOD（P<0.05）. It also showed inhibitory effect on the NF-κB protein expression（P<0.05）. Conclusion Polygonatum sibiricum polysaccharide has antioxidant effect in vitro. Its inhibitory effect on mice inflammatory bowel disease may be the inhibition pathway of NF-κB， which is achieved by reducing the MDA， MPO production and increasing the SOD content. [參考文献]
[1] 武晓琳，赵亚楠，王丽波，等. 炎症性肠病相关信号通路的研究进展[J]. 国际消化病杂志，2016，36（2）：105-107.
[2] Yang Y，Bazhin AV，Werner J，et al. Reactive oxygen species in the immune system[J]. Int Rev Immunol，2013， 32（3）：249-270.
[3] 段华，王保，张跃文.黄精多糖对肝癌H22 移植瘤小鼠的抑瘤作用及机制研究[J]. 中药新药与临床药理研究， 2014，25（1）：5-7.
[4] 李友元，邓洪波，张萍，等.黄精多糖降脂及抗动脉粥样硬化的实验研究[J]. 中国动脉硬化杂志，2005，13（4）： 429-431.
[5] 雷升萍，王靓，龙子江，等. 黄精多糖通过TLR4-MyD88-NF-κB通路抑制缺氧/复氧H9C2心肌细胞炎性因子释放[J]. 中国药理学通报，2017，33（2）：255-260.
[6] Jin M，Cai YX，Li JR，et al. 10-Phennanthroline-Fe2 oxidative assay of hydroxyl radical produced by H2O2/Fe2 [J].Prog Biochem Biophys，1996，23（6）：553-556.
[7] 周海玲，马麟，易智彪. 基于三种体外抗氧化方法对白木香种子抗氧化能力的研究[J]. 中国医药导报，2016， 13（22）：12-15.
[8] Fonseca SF，Padilha NB，Thurow S，et al. Ultrasound-promoted copper-catalyzed synthesis of bis-arylselanyl chrysin derivatives with boosted antioxidant and anticancer activities[J]. Ultrason Sonochem，2017，39：827-836.
[9] Daniela Impellizzeri，Giuseppe Bruschetta，Rosanna Di Paola，et al. The anti-inflammatory and antioxidant effects of bergamot juice extract（BJe）in an experimental model of inflammatory bowel disease[J]. Clinical Nutrition，2015，34（6）：1146-1154.
[10] Pallio G，Bitto A，Pizzino G，et al. Adenosine receptor stimulation by polydeoxyribonucleotide improves tissue repair and symptomology in experimental colitis[J]. Front Pharmacol，2016，（7）：273.
[11] Tanida S，Mizoshita T，Mizushima T，et al. Involvement of oxidative stress and mucosal addressin cell adhesion molecule-1（MAdCAM-1）in inflammatory bowel disease[J].J Clin Biochem Nutr，2011，48（2）：112-116.
[12] Pravda J. Radical induction theory of ulcerative colitis[J].World J Gastroenterol，2005，11（16）：2371-2384.
[13] Pavlick KP，Laroux FS，Fuseler J，et al. Role of reactive metabolites of oxygen and nitrogen in inflammatory bowel disease[J]. Free Radic Biol Med，2002，33（3）：311-322.
[14] Hatsugai M，Kurokawa MS，Kouro T，et al. Protein profiles of peripheral blood mononuclear cells are useful for differential diagnosis of ulcerative colitis and Crohn’s disease[J]. J Gastroenterol，2010，45（5）：488-500.
[15] Cracowski JL，Bonaz B，Bessard G，et al. Increased urinary F2-isoprostanes in patients with Crohn’s disease[J]. Am J Gastroenterol，2002，97（7）：99-103.
[16] Tuzun A，Erdil A，Inal V，et al. Oxidative stress and antioxidant capacity in patients with inflammatory bowel disease[J]. Clin Biochem，2002，35（7）：569-572.
[17] 王皓，欧阳欽，胡仁伟. 二硝基苯磺酸结肠炎动物模型的建立[J]. 胃肠病学，2001，6（1）：7-10.
[18] Michiels C，Minet E，Mottet D，et al. Regulation of gene expression by oxygen：NF-kappa B and HIF-1，two extremes[J]. Free RadicBiol Med，2002，33（9）：1231-1242.
[19] Ghosh S，Hayden MS. New regulators of NF-kappaB in inflammation[J]. Nat Rev Immunol，2008，8（11）：837-848.
[20] Wang K，Wan YJ. Nuclear receptors and inflammatory diseases[J]. ExP Biol Med（Maywood），2008，233（5）：496-506.
[21] Wahli W. A gut feeling of the PXR，PPAR and NF-kappaB connection[J]. J Intern Med，2008，263（6）：613-619.
（收稿日期：2017-08-31）
[关键词] 黄精多糖；炎症性肠病；体外抗氧化能力；NF-κB通路
[中图分类号] R285 [文献标识码] A [文章编号] 1673-9701（2017）29-0027-04
Study on the antioxidant effect in vitro of polygonatum sibiricum polysaccharides and its effect on inflammatory bowel disease in mice
XUE Xuebin1 FANG Shuhua2 WANG Huajun3
1.Department of Drug and Equipment， Zhenjiang Second People’s Hospital in Jiangsu Province， Zhenjiang 212000， China； 2.Department of Pharmacy， Zhenjiang First People’s Hospital in Jiangsu Province， Zhenjiang 212000， China； 3.Department of Pharmacy， the Affiliated Hospital of Jiangsu University， Zhenjiang 212001， China
[Abstract] Objective To investigate the anti-oxidative effects of polygonatum sibiricum polysaccharides（PSP） in vitro and its protective effect on inflammatory bowel disease in mice. Methods In vitro anti-oxidative experiment was carried out to observe the effect of PSP on the generation of hydroxyl radical（·OH）， the effect of ·OH on the destruction of erythrocytes and whether lipid production of liver homogenate had inhibitory effect. The model of mice inflammatory bowel disease was established and was given PSP treatment. Mice disease activity score（DAI） was calculated， MDA， SOD and MPO in colonic tissue of mice were detected， and the expression of NF-κB protein in the colonic tissue of mice was determined. Results PSP showed inhibitory effect on the generation of ·OH in vitro and ·OH showed inhibitory effect on the destruction of erythrocytes（P<0.01）， which inhibited liver homogenate lipid peroxidation process（P<0.01）， and it’s in vitro anti-oxidant effect was dose-dependent（P<0.05-0.01）. Compared with the model group， PSP can reduce the DAI score（P<0.01）， inhibit the generation of MDA and MPO in colonic tissues（P<0.05-0.01）， and increase the content of SOD（P<0.05）. It also showed inhibitory effect on the NF-κB protein expression（P<0.05）. Conclusion Polygonatum sibiricum polysaccharide has antioxidant effect in vitro. Its inhibitory effect on mice inflammatory bowel disease may be the inhibition pathway of NF-κB， which is achieved by reducing the MDA， MPO production and increasing the SOD content. [參考文献]
[1] 武晓琳，赵亚楠，王丽波，等. 炎症性肠病相关信号通路的研究进展[J]. 国际消化病杂志，2016，36（2）：105-107.
[2] Yang Y，Bazhin AV，Werner J，et al. Reactive oxygen species in the immune system[J]. Int Rev Immunol，2013， 32（3）：249-270.
[3] 段华，王保，张跃文.黄精多糖对肝癌H22 移植瘤小鼠的抑瘤作用及机制研究[J]. 中药新药与临床药理研究， 2014，25（1）：5-7.
[4] 李友元，邓洪波，张萍，等.黄精多糖降脂及抗动脉粥样硬化的实验研究[J]. 中国动脉硬化杂志，2005，13（4）： 429-431.
[5] 雷升萍，王靓，龙子江，等. 黄精多糖通过TLR4-MyD88-NF-κB通路抑制缺氧/复氧H9C2心肌细胞炎性因子释放[J]. 中国药理学通报，2017，33（2）：255-260.
[6] Jin M，Cai YX，Li JR，et al. 10-Phennanthroline-Fe2 oxidative assay of hydroxyl radical produced by H2O2/Fe2 [J].Prog Biochem Biophys，1996，23（6）：553-556.
[7] 周海玲，马麟，易智彪. 基于三种体外抗氧化方法对白木香种子抗氧化能力的研究[J]. 中国医药导报，2016， 13（22）：12-15.
[8] Fonseca SF，Padilha NB，Thurow S，et al. Ultrasound-promoted copper-catalyzed synthesis of bis-arylselanyl chrysin derivatives with boosted antioxidant and anticancer activities[J]. Ultrason Sonochem，2017，39：827-836.
[9] Daniela Impellizzeri，Giuseppe Bruschetta，Rosanna Di Paola，et al. The anti-inflammatory and antioxidant effects of bergamot juice extract（BJe）in an experimental model of inflammatory bowel disease[J]. Clinical Nutrition，2015，34（6）：1146-1154.
[10] Pallio G，Bitto A，Pizzino G，et al. Adenosine receptor stimulation by polydeoxyribonucleotide improves tissue repair and symptomology in experimental colitis[J]. Front Pharmacol，2016，（7）：273.
[11] Tanida S，Mizoshita T，Mizushima T，et al. Involvement of oxidative stress and mucosal addressin cell adhesion molecule-1（MAdCAM-1）in inflammatory bowel disease[J].J Clin Biochem Nutr，2011，48（2）：112-116.
[12] Pravda J. Radical induction theory of ulcerative colitis[J].World J Gastroenterol，2005，11（16）：2371-2384.
[13] Pavlick KP，Laroux FS，Fuseler J，et al. Role of reactive metabolites of oxygen and nitrogen in inflammatory bowel disease[J]. Free Radic Biol Med，2002，33（3）：311-322.
[14] Hatsugai M，Kurokawa MS，Kouro T，et al. Protein profiles of peripheral blood mononuclear cells are useful for differential diagnosis of ulcerative colitis and Crohn’s disease[J]. J Gastroenterol，2010，45（5）：488-500.
[15] Cracowski JL，Bonaz B，Bessard G，et al. Increased urinary F2-isoprostanes in patients with Crohn’s disease[J]. Am J Gastroenterol，2002，97（7）：99-103.
[16] Tuzun A，Erdil A，Inal V，et al. Oxidative stress and antioxidant capacity in patients with inflammatory bowel disease[J]. Clin Biochem，2002，35（7）：569-572.
[17] 王皓，欧阳欽，胡仁伟. 二硝基苯磺酸结肠炎动物模型的建立[J]. 胃肠病学，2001，6（1）：7-10.
[18] Michiels C，Minet E，Mottet D，et al. Regulation of gene expression by oxygen：NF-kappa B and HIF-1，two extremes[J]. Free RadicBiol Med，2002，33（9）：1231-1242.
[19] Ghosh S，Hayden MS. New regulators of NF-kappaB in inflammation[J]. Nat Rev Immunol，2008，8（11）：837-848.
[20] Wang K，Wan YJ. Nuclear receptors and inflammatory diseases[J]. ExP Biol Med（Maywood），2008，233（5）：496-506.
[21] Wahli W. A gut feeling of the PXR，PPAR and NF-kappaB connection[J]. J Intern Med，2008，263（6）：613-619.
（收稿日期：2017-08-31）