溃结爽灌肠组方不同组分对实验性溃疡性结肠炎大鼠的氧化应激及能量代谢水平的影响
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摘要
目的:分别从氧化应激及能量代谢角度研究溃结爽灌肠方对实验性溃疡性结肠炎(UC)大鼠的作用及其疗效机制,并应用正交试验设计,分析溃结爽灌肠方中各组分在不同水平对UC大鼠的的单因素效应及量效关系。方法:按照L9(34)正交表设计,70只清洁级雄性wistar大鼠进行9组试验。葡聚糖硫酸钠(DSS)法复制UC大鼠模型后,以溃结爽方灌肠10 d,麻醉取材,计算结肠黏膜损伤评分(CMDI),制作病理切片;生化法检测大鼠结肠黏膜谷胱甘肽(GSH)、超氧化物歧化酶(SOD)活性及丙二醛(MDA)、腺苷三磷酸(ATP)、一氧化氮(NO)表达水平。结果:模型组抗氧化因子SOD、GSH活性明显降低,脂质过氧化产物MDA及NO水平显著增高,并且能量代谢受到影响,ATP水平降低,与空白组比差异有显著性(P<0.05)。溃结爽灌肠能够不同程度改善UC大鼠氧化应激损伤及能量代谢水平,提高SOD、GSH活性及ATP水平,降低MDA、NO的表达,并且以B组效应最佳(P<0.05)。在结肠黏膜损伤评分(CMDI)方面,A组B组C组E组F组I组均较模型组有不同程度改善(P<0.05),结果与抗氧化因子活性(SOD、GSH)及能量代谢(ATP酶)表达水平变化具有一致性。4味中药黄柏、苦参、白及、地榆在高、中、低剂量均可不同程度提高抗氧化因子SOD、GSH活性及ATP的表达,同时降低大鼠结肠黏膜MDA、NO的含量,但随剂量变化其表达水平呈现不同变化趋势。结论:溃结爽灌肠方可以明显降低UC模型大鼠结肠黏膜氧化应激损伤,提高能量代谢水平,从而减轻UC大鼠肠道炎症损伤,促进肠道黏膜修复。
Objective: To study the effect and the therapeutic mechanism of Kui Jie Shuang enema on experimental ulcerative colitis(UC) rats from the angle of oxidative stress and energy metabolism,and to analyze the effect of the single factor effect and the doseeffect relationship on the UC rats at different levels by orthogonal design. Methods: according to L9(34) orthogonal design,70 clean male Wistar rats were tested in 9 groups. After replicating the rat model of ulcerative colitis with dextran sodium sulfate(DSS) method,Kui Jie Shuang was used for enema for 10 d; materials was selected by anesthesia,the colonic mucosa injury score(CMDI) was calculated,and the pathological sections were made. The biochemical method was used to detect the glutathione(GSH),the activity of superoxide dismutase(SOD),and the expression levels of malondialdehyde(MDA),adenosine triphosphate(ATP) and nitric oxide(NO) in the rat's colon mucosa. Results: the activity of antioxidant factor SOD and GSH decreased significantly in the model group,the level of MDA and NO increased significantly,the energy metabolism was influenced,the level of ATP decreased,and the difference was significant(P < 0. 05). Kui Jie Shuang enema could improve the oxidative stress injury and energy metabolism of UC rats in different degrees,improve the activity of SOD and GSH and the level of ATP,and reduce the expression of MDA and NO,with the best effect in group B(P < 0. 05). Compared to model group,CMDI in group A,B,C,E,F and I had different degrees of improvement(P < 0. 05),and the result was consistent with the changes of expression level of activity of SOD and GSH and energy ATP expression. The 4 Chinese herbs,Huangbai,Kushen,Baiji and Diyu of different doses could increase the expression of SOD,GSH activity and ATP expression,and reduce the content of MDA and NO in the colon mucosa of rats,but the expression level varied with dose.Conclusion: Kui Jie Shuang enema can significantly reduce the oxidative stress damage in the colon mucosa of UC model rats and improve the level of energy metabolism,thus reducing the damage of intestinal inflammation and promoting the repair of intestinal mucosa.
Objective: To study the effect and the therapeutic mechanism of Kui Jie Shuang enema on experimental ulcerative colitis(UC) rats from the angle of oxidative stress and energy metabolism,and to analyze the effect of the single factor effect and the doseeffect relationship on the UC rats at different levels by orthogonal design. Methods: according to L9(34) orthogonal design,70 clean male Wistar rats were tested in 9 groups. After replicating the rat model of ulcerative colitis with dextran sodium sulfate(DSS) method,Kui Jie Shuang was used for enema for 10 d; materials was selected by anesthesia,the colonic mucosa injury score(CMDI) was calculated,and the pathological sections were made. The biochemical method was used to detect the glutathione(GSH),the activity of superoxide dismutase(SOD),and the expression levels of malondialdehyde(MDA),adenosine triphosphate(ATP) and nitric oxide(NO) in the rat's colon mucosa. Results: the activity of antioxidant factor SOD and GSH decreased significantly in the model group,the level of MDA and NO increased significantly,the energy metabolism was influenced,the level of ATP decreased,and the difference was significant(P < 0. 05). Kui Jie Shuang enema could improve the oxidative stress injury and energy metabolism of UC rats in different degrees,improve the activity of SOD and GSH and the level of ATP,and reduce the expression of MDA and NO,with the best effect in group B(P < 0. 05). Compared to model group,CMDI in group A,B,C,E,F and I had different degrees of improvement(P < 0. 05),and the result was consistent with the changes of expression level of activity of SOD and GSH and energy ATP expression. The 4 Chinese herbs,Huangbai,Kushen,Baiji and Diyu of different doses could increase the expression of SOD,GSH activity and ATP expression,and reduce the content of MDA and NO in the colon mucosa of rats,but the expression level varied with dose.Conclusion: Kui Jie Shuang enema can significantly reduce the oxidative stress damage in the colon mucosa of UC model rats and improve the level of energy metabolism,thus reducing the damage of intestinal inflammation and promoting the repair of intestinal mucosa.
引文
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[27]贺国芳,丁伊玲,徐清霞,等.白及多糖对小鼠急性酒精性肝损伤的保护作用[J].中国医院药学杂志,2015,35(18):1 658-1 661
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[32]周承志,姜楠,周从辉,等.复方苦参汤对溃疡性结肠炎模型大鼠的干预及作用机制研究[J].中国药师,2016,19(10):1 816-1 820
[2]胡品津.炎症性肠病诊断与治疗的共识意见(2012年·广州)解读[J],胃肠病学,2012,17(12):709-711
[3]Almeer RS1,Mahmoud SM2,Amin HK3.Ziziphus spinachristi fruit extract suppresses oxidative stress and p38 MAPK expression in ulcerative colitis in rats via induction of Nrf2 and HO-1 expression[J].Food Chem Toxicol,2018,115(5):49-62
[4]Lee SH,Kwon JE,Cho ML1.Immunological pathogenesis of inflammatory bowel disease[J].Intest Res,2018,16(1):26-42
[5]Shouval DS,Rufo PA.The Role of Environmental Factors in the Pathogenesis of Inflammatory Bowel Diseases:A Review[J].JAMA Pediatr,2017,171(10):999-1 005
[6]Mrowicka M1,Mrowicki J1,Mik M2,et al.Association between SOD1,CAT,GSHPX1 polymorphisms and the risk of inflammatory bowel disease in the Polish population[J].Oncotarget,2017,8(65):109 332-109 339
[7]Shalkami AS1,Hassan M1,Bakr AG1.Anti-inflammatory,antioxidant and anti-apoptotic activity of diosmin in acetic acid-induced ulcerative colitis[J].Hum Exp Toxicol,2018,37(1):78-86
[8]Formentini L,Santacatterina F,Núez de Arenas C,et al.Mitochondrial ROS Production Protects the Intestine from Inflammation through Functional M2 Macrophage Polarization[J].Cell Rep,2017,19(6):1 202-1 213
[9]Patlevi8 P1,Va3kováJ2,3vorc P Jr3,et al.Reactive oxygen species and antioxidant defense in human gastrointestinal diseases[J].Integr Med Res,2016,5(4):250-258
[10]Sakthivel KM1,Guruvayoorappan C.Amentoflavone inhibits i NOS,COX-2 expression and modulates cytokine profile,NF-κB signal transduction pathways in rats with ulcerative colitis[J].Int Immunopharmacol,2013,17(3):907-16
[11]Chu XQ,Wang J,Chen GX,et al.Overexpression of microRNA-495 improves the intestinal mucosal barrier function by targeting STAT3 via inhibition of the JAK/STAT3 signaling pathway in a mouse model of ulcerative colitis[J].Pathol Res Pract,2018,214(1):151-162
[12]El-Kheshen G,Moeini M,Saadat M.Susceptibility to Ulcerative Colitis and Genetic Polymorphisms of A251G SOD1 and C-262T CAT[J].J Med Biochem,2016,35(3):333-336
[13]Kruidenier L,Kuiper,Lamers CB,et al.Intestinal oxidative damage in inflammatory bowel disease:semi-quantification,localization,and association with mucosal antioxidants[J].J Pathol,2003,201(1):28-36
[14]Zhang Z,Yang L,Wang B,et al.Protective role of liriodendrin in mice with dextran sulphate sodium-induced?ulcerative colitis[J].Int Immunopharmacol,2017 Nov,52:203-210
[15]Shao J,Liu Z,Wang L,et al.Screening of the optimized prescription from Suqingwan in terms of its therapeutic effect on DSS-induced ulcerative colitis by its regulation of inflammatory and oxidative mediators[J].J Ethnopharmacol,2017,202(4):54-62
[16]Akazawa Y,Kubo M,Zhang R,et al.Inhibition of arginase ameliorates experimental ulcerative colitis in mice[J].Free Radic Res,2013,47(3):137-45
[17]Nguyen K,Garcia A,Sani MA,et al.Interaction of N-terminal peptide analogues of the Na+,K+-ATPase?with membranes[J].Biochim Biophys Acta,2018,1860(6):1 282-1 291
[18]邱林,李美红,夏洽思,等.中医药对线粒体保护机制的研究进展[J].湖南中医杂志,2017,33(6):214-215
[19]温超玮,仇如意,胡念琪,等.线粒体DNA单体型M8a对转线粒体细胞线粒体能量代谢的影响[J].中国细胞生物学学报,2017,39(5):607-614
[20]Jin X,Chen D1,Zheng RH,et al.miRNA-133a-UCP2pathway regulates inflammatory bowel disease progress by influencing inflammation,oxidative stress and energy metabolism[J].World J Gastroenterol,2017,23(1):76-86
[21]吴嘉瑞,张冰,张光敏.黄柏药理作用研究进展[J].亚太传统医药,2009,5(11):160-162
[22]Chen YY,Li RY,Shi MJ,et al.Demethyleneberberine alleviates inflammatory bowel disease in mice through regulating NF-κB signaling and T-helper cell homeostasis[J].Inflamm Res,2017,66(2):187-196
[23]徐珊,张凡,刘蓬蓬.基于大鼠物质、能量代谢研究炮制对黄柏药性的影响[J].中药材,2015,38(9):1 835-1 841
[24]代良敏,熊永爱,范奎,等.地榆化学成分与药理作用研究进展[J].中国实验方剂学杂志,2016,22(20):189-195
[25]Jang E,Inn KS,Jang YP,et al.Phytotherapeutic Activities of?Sanguisorba officinalis and its Chemical Constituents:A Review[J].Am J Chin Med,2018,12:1-20
[26]孙爱静,庞素秋,王国权.中药白及化学成分与药理活性研究进展[J].环球中医药,2016,9(4):507-511
[27]贺国芳,丁伊玲,徐清霞,等.白及多糖对小鼠急性酒精性肝损伤的保护作用[J].中国医院药学杂志,2015,35(18):1 658-1 661
[28]Fan H,Liu XX,Zhang LJ,et al.Intervention effects of QRZSLXF,a Chinese medicinal herb recipe,on the DOR-beta-arrestin1-Bcl2 signal transduction pathway in a rat model of ulcerative colitis[J].J Ethnopharmacol,2014,154(1):88-97
[29]王悦,姜雪,丁菲,等.中药苦参药理作用及应用研究进展[J].山东化工,2017,46(15):66-69
[30]Fu X,Sun F,Wang F,et al.Aloperine Protects Mice against DSS-Induced Colitis by PP2A-Mediated PI3K/Akt/m TOR Signaling Suppression[J].Mediators Inflamm,2017,20(17):5 706 152
[31]张顺景,吕芳.苦参对大鼠溃疡性结肠炎中SOD、MDA表达的影响[J].包头医学院学报,2005,21(2):112-113
[32]周承志,姜楠,周从辉,等.复方苦参汤对溃疡性结肠炎模型大鼠的干预及作用机制研究[J].中国药师,2016,19(10):1 816-1 820