马齿苋-肉桂联用提取工艺及其对DSS诱导溃疡性结肠炎的作用研究
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摘要
目的:对马齿苋-肉桂联用最佳配比和多糖提取工艺及其多糖抗实验性溃疡性结肠炎(UC)作用进行研究。方法:采用L_9(3~4)正交试验对马齿苋-肉桂联用最佳配比以及多糖的提取工艺进行优化;用3%葡聚糖硫酸钠(DSS)诱导小鼠UC模型,灌胃300 mg·kg~(-1)柳氮磺胺吡啶为阳性对照,马齿苋-肉桂低中高实验组分别以100、200、400 mg·kg~(-1)的多糖水溶液灌胃治疗,观察小鼠给药前后的体征、结肠组织形态、血清白细胞介素-10(IL-10)及肿瘤坏死因子(TNF-α)的含量变化。结果:最佳提取工艺为马齿苋-肉桂12∶5,料液比1∶30,90℃提取5 h;给药治疗UC小鼠后,HE染色显示小鼠结肠黏膜上皮溃疡处有不同程度修复;给药组小鼠血清抑炎因子IL-10含量较模型组显著上升(P<0.01),促炎因子TNF-α含量较模型组显著降低(P<0.01)。结论:本实验得出马齿苋配伍肉桂联用最佳配比和最佳提取工艺。给药治疗UC小鼠后,从疾病活动指数(DAI)评分、HE染色、IL-10及TNF-α含量反映,马齿苋-肉桂联用对UC有治疗作用,为临床治疗UC提供了新思路。
Objective:To explore the optimal ratio of Portulaca oleracea combing with Cinnamomm cassia and evaluate their protecitive against DSS induced ulcerative colitis.Methods:Orthogonal experiment design L_9(3~4) was employed to optimize the extraction process.UC mice were induced by 3% DSS and the DAI,H&E staining were enrolled to evaluated the protective effect of the extraction.The expression levels of IL-10 and TNF-α were also detected.Results:The best ratio of P.oleracea combing with C.cassia was 12:5.The extraction process was verified at 90 ℃ for 5 h.After administration of the extraction,the morphological structure of UC was improved and the inflammatory cells were almost disappeared,indicating the satisfied therapeutic effect.The expression level of IL-10 was greatly increased,while the pro-inflammatory cytokine TNF-α was decreased significantly.Conclusion:P.oleracea combing with C.cassia could improve the health conditions of UC mice via regulating the levels of inflammatory cytokines.And the extraction also can improve the morphological structure of UC.
Objective:To explore the optimal ratio of Portulaca oleracea combing with Cinnamomm cassia and evaluate their protecitive against DSS induced ulcerative colitis.Methods:Orthogonal experiment design L_9(3~4) was employed to optimize the extraction process.UC mice were induced by 3% DSS and the DAI,H&E staining were enrolled to evaluated the protective effect of the extraction.The expression levels of IL-10 and TNF-α were also detected.Results:The best ratio of P.oleracea combing with C.cassia was 12:5.The extraction process was verified at 90 ℃ for 5 h.After administration of the extraction,the morphological structure of UC was improved and the inflammatory cells were almost disappeared,indicating the satisfied therapeutic effect.The expression level of IL-10 was greatly increased,while the pro-inflammatory cytokine TNF-α was decreased significantly.Conclusion:P.oleracea combing with C.cassia could improve the health conditions of UC mice via regulating the levels of inflammatory cytokines.And the extraction also can improve the morphological structure of UC.
引文
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[19]冯津津.马齿苋的化学成分及药理作用研究进展[J].云南中医中药杂志,2013,34(7):66-68.
[20]冯澜,李绍民,代立娟,等.马齿苋多糖对溃疡性结肠炎小鼠肠黏膜细胞因子及肠道菌群的影响[J].中国微生态学杂志,2015,27(2):139-142.
[21]赵月.肉桂黄酮类抗氧化活性成分的HPLC法识别及其成分分析[D].天津:天津商业大学,2017.
[2]BERNSTEIN C N,FRIED M,KRABSHUIS J H,et al.World Gastroenterology Organization Practice Guidelines for the diagnosis and management of IBD in 2010[J].Inflamm Bowel Dis,2010,16(1):112-124.
[3]CAANA C Sristina,NEAGOE I B,COZMA V,et al Contribution of the IL-17/IL-23 axis to the pathogenesis of inflammatory bowel disease[J].World J Gastroentero,2015,21(19):5823-5830.
[4]LI J,HUANG Y Q,HUANG C,et al.Plasma miR-92a as a promi sing predictive biomarker for arteriosclerosis in patients with essential hypertension[J].S Chin J Cariol,2016,17(4):196-203.
[5]柳亦松,唐昭山,刘兆颖,等.血根碱对大鼠结肠炎及炎症通路中相关因子的影响[J].中国现代中药,2017,19(10):43-48.
[6]王晓妍,孙蓉,张小平,等.四君痛泻方对溃疡性结肠炎患者血清NGF、TNF-α、IL-10的影响[J].河南中医,2018,38(4):578-581.
[7]汪荔.马齿苋多糖的提取分离及其抗溃疡性结肠炎作用机理研究[D].咸阳:陕西中医药大学,2017.
[8]张超.基于高通量测序研究肉桂精油对肠炎小鼠肠道菌群的影响[D].哈尔滨:东北农业大学,2017.
[9]郭志烨,韩丽,杨明,等.中药多糖定量测定方法的探讨[J].中成药,2014,36(10):2172-2176.
[10]刘瑞江,张业旺,闻崇炜,等.正交试验设计和分析方法研究[J].实验技术与管理,2010,27(9):52-55.
[11]魏永凯,耿福能,赵微,等.美沙拉嗪、柳氮磺胺吡啶对葡聚糖硫酸钠诱导的Balb/c小鼠急性溃疡性结肠炎的治疗和免疫影响[J].中国医院药学杂志,2016,36(14):1190-1195.
[12]衡宇,李晰,孙涛,等.葡聚糖硫酸钠自由饮用与灌胃诱导小鼠溃疡性结肠炎模型的对比研究[J].中国药师,2017,20(4):603-606.
[13]MURANO M,MAEMURA K,HIRATA I,et al.Therapeutic effect of intracolonically administered nuclear factor kappa B(p65)antisense oligonucleotide on mouse dextran sulphate sodium(DSS)-induced colitis.[J].Clin Exp Immunol,2000,120(1):51-58.
[14]SA'NCHEZ-FIDALGO S,CA'RDENO A,SA'NCHEZ-HIDALGOM,et al.Dietary extra virgin olive oil polyphenols supplementation modulates DSS-induced chronic colitis in mice[J].J Nutr Biochem,2013,24(7):1401-1413.
[15]孔鹏飞,赵兵,覃勤,等.葡聚糖硫酸钠致大鼠急性溃疡性结肠炎模型建立与评价[J].中华结直肠疾病电子杂志,2015,4(6):617-619.
[16]周正,马婷,冯澜,等.马齿苋多糖对溃疡性结肠炎小鼠肠道菌群及血内毒素的影响[J].中国微生态学杂志,2014,26(6):646-648.
[17]陈晓杨.芍药汤治疗溃疡性结肠炎36例临床观察[J].实用中医内科杂志,2014,28(5):39-40.
[18]SHEN H,TANG G,ZENG G,et al.Purification and characterization of an antitumor polysaccharide from Portulaca oleracea L[J].Carbohydr Polym,2013,93(2):395-400.
[19]冯津津.马齿苋的化学成分及药理作用研究进展[J].云南中医中药杂志,2013,34(7):66-68.
[20]冯澜,李绍民,代立娟,等.马齿苋多糖对溃疡性结肠炎小鼠肠黏膜细胞因子及肠道菌群的影响[J].中国微生态学杂志,2015,27(2):139-142.
[21]赵月.肉桂黄酮类抗氧化活性成分的HPLC法识别及其成分分析[D].天津:天津商业大学,2017.
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