异功散通过抑制炎症反应促进慢性病贫血小鼠脾组织结构修复研究
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摘要
目的观察异功散通过抑制炎症反应,对慢性病贫血小鼠脾组织结构修复的影响。方法将60只8周龄雄性C57 BL/6小鼠随机分为空白对照组、模型组、异功散组,每组20只。除空白对照组外,其余各组复制慢性病贫血小鼠模型(第1天腹腔注射脂多糖,第7天腹腔注射酵母糖A)。注射酵母糖A后第5天,异功散组灌胃异功散水煎液(15.413 g·kg~(-1)·d~(-1)),空白对照组、模型组灌胃0.2 ml超纯水。各组分别于连续干预1周和2周后采集全血,自动细胞计数仪检测白细胞计数、中性粒细胞计数及血红蛋白水平;分离脾组织,HE染色观察脾组织结构,电镜观察小鼠脾脏线粒体情况。结果与空白对照组比较,模型组白细胞及中性粒细胞计数升高(P<0.01),血红蛋白水平下降(P<0.01);脾脏明显增大,脾脏指数增加(P<0.01),脾组织线粒体正常结构消失。连续干预1周和2周后,与模型组比较,异功散组小鼠白细胞及中性粒细胞计数明显下降(P<0.01),血红蛋白水平明显升高(P<0.01);脾脏均明显缩小,脾脏指数降低(P<0.01);电镜下观察到小鼠脾脏线粒体结构接近正常,受损线粒体数量减少。结论异功散可通过抑制炎症反应,减少炎症因子对脾组织结构的损伤,进而改善慢性病贫血小鼠的贫血状况。
Objective To observe the effect of Yigong Powder on repair of spleen tissue structure by inhibiting inflammatory reactions in mice with anemia of chronic disease(ACD). Methods Sixty 8-week-old male C57 BL/6 mice were randomly assigned into blank control group, model group and Yigong Powder group, 20 mice in each group. Except the blank control group, the model of mice with ACD was copied in the other groups(lipopolysaccharide and zymosan A were injected intraperitoneally at day 1 and day 7 respectively). At 5 days after injection of zymosan A, Decoction of Yigong Powder(15.413 g·kg~(-1)·d~(-1)) was administered by gavage to mice in the Yigong Powder group and 0.2 ml of ultra-pure water was given by gavage to the blank control and model groups. After 1 week and 2 weeks of treatment, whole blood was collected, white blood cell(WBC) count, neutrophil count and hemoglobin level were determined with an automated cell counter; the spleen tissue was isolated to observe the structure by HE staining; and mitochondria of spleen was observed under electron microscope. Results Compared to the blank control group, WBC count and neutrophil count increased and the hemoglobin level decreased(P<0.01), the spleen enlarged significantly, the spleen index increased(P<0.01), and the normal structure of mitochondria disappeared in spleen tissue in the model group. After 1 week and 2 weeks of treatment, compared to the model group,WBC count and neutrophil count decreased significantly and the hemoglobin level increased significantly( P<0.01); the spleen shrank significantly( P<0.01), and the spleen index decreased( P<0.01) in the Yigong Powder group. The mitochondrial structure was observed to be close to normal under electron microscope,and the number of damaged mitochondria was reduced. Conclusion Yigong Powder improves anemia in mice with ACD by inhibiting inflammatory reactions and reducing damage to the spleen tissue structure caused by inflammatory factors.
Objective To observe the effect of Yigong Powder on repair of spleen tissue structure by inhibiting inflammatory reactions in mice with anemia of chronic disease(ACD). Methods Sixty 8-week-old male C57 BL/6 mice were randomly assigned into blank control group, model group and Yigong Powder group, 20 mice in each group. Except the blank control group, the model of mice with ACD was copied in the other groups(lipopolysaccharide and zymosan A were injected intraperitoneally at day 1 and day 7 respectively). At 5 days after injection of zymosan A, Decoction of Yigong Powder(15.413 g·kg~(-1)·d~(-1)) was administered by gavage to mice in the Yigong Powder group and 0.2 ml of ultra-pure water was given by gavage to the blank control and model groups. After 1 week and 2 weeks of treatment, whole blood was collected, white blood cell(WBC) count, neutrophil count and hemoglobin level were determined with an automated cell counter; the spleen tissue was isolated to observe the structure by HE staining; and mitochondria of spleen was observed under electron microscope. Results Compared to the blank control group, WBC count and neutrophil count increased and the hemoglobin level decreased(P<0.01), the spleen enlarged significantly, the spleen index increased(P<0.01), and the normal structure of mitochondria disappeared in spleen tissue in the model group. After 1 week and 2 weeks of treatment, compared to the model group,WBC count and neutrophil count decreased significantly and the hemoglobin level increased significantly( P<0.01); the spleen shrank significantly( P<0.01), and the spleen index decreased( P<0.01) in the Yigong Powder group. The mitochondrial structure was observed to be close to normal under electron microscope,and the number of damaged mitochondria was reduced. Conclusion Yigong Powder improves anemia in mice with ACD by inhibiting inflammatory reactions and reducing damage to the spleen tissue structure caused by inflammatory factors.
引文
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[14]谢坤莹,魏锦.炎症因子在RA合并慢性病贫血发病过程的相关性研究[J].世界最新医学信息文摘,2017,17(40):17-19.
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[16]罗梅宏.从脾主运化和运脾生血理论探讨慢性病贫血的中医病机和治疗[J].中医杂志,2013,54(18):1556-1557.
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[19]LIU X B,NGUYEN N B,MARQUESS K D,et al.Regulation of hepcidin and ferroportin expression by lipopolysaccharide in splenic macrophages[J].Blood Cells Mol Dis,2005,35(1):47-56.
[20]GARDENGHI S,RENAUD T M,MELONI A,et al.Distinct roles for hepcidin and interleukin-6 in the recovery from anemia in mice injected with heat-killed Brucella abortus[J].Blood,2014,123(8):1137-1145.
[21]KIM A,FUNG E,PARIKH S G,et al.A mouse model of anemia of inflammation:complex pathogenesis with partial dependence on hepcidin[J].Blood,2014,123(8):1129-1136.
[22]张引红.补骨脂素对类风湿性关节炎小鼠模型的免疫调节作用[J].中国实验动物学报,2017,25(2):207-210.
[23]张梦思.当归多糖与阿糖胞苷联合注射对人白血病模型小鼠脾脏结构与功能的影响[J].中药药理与临床,2014,30(4):62-66.
[24]ZHONG Z,SANCHEZLOPEZ E,KARIN M,et al.Autophagy,NLRP3inflammasome and auto-inflammatory/immune diseases[J].Clin Exp Rheumatol,2016,34(4 Suppl 98):12-16.
[25]林妘.NF-κB通过清除受损线粒体抑制炎症小体的活化[J].中医肿瘤生物杂志,2016,164(5):896-910.
[26]周露露,牛佳丽,陈陶陶,等.高浓度LPS损伤巨噬RAW264.7线粒体功能的研究[J].吉林医药学院学报,2015,36(1):17-18.
[2]NAIRZ M,THEURL I,WOLF D,et al.Iron deficiency or anemia of inflammation?[J].Wien Med Wochenschr,2016,166(13-14):411-423.
[3]WARDROP S L,RICHARDSON D R.Interferon-γand lipopolysaccharide regulate the expression of Nramp2 and increase the uptake of iron from low relative molecular mass complexes by macrophages[J].Eur J Biochem,2010,267(22):6586-6593.
[4]DELGADO-VEGA A M,ALARCON-RIQUELME M E,KOZYREV SV,et al.Genetic associations in type I interferon related pathways with autoimmunity[J].Arthritis Res Ther,2010(12 Suppl 1):S2.
[5]WEISS G,GOODNOUGH L T,LAWRENCE T,et al.Anemia of chronic disease[J].N Engl J Med,2005,352(10):1011-1023.
[6]MENG Z S,JIA H Y,WU X Q,et al.Iron metabolism and overload[J].Med Recapitulate,2012,18(1):43-46.
[7]RAMM G A,RUDDELL R G.Iron homeostasis,hepatocellular injury,and fibrogenesis in hemochromatosis:the role of inflammation in a noninflammatory liver disease[J].Semin Liver Dis,2010,30(3):271-287.
[8]郑秦,管宇,王志成,等.异功散对脂多糖介导的小鼠铁代谢紊乱的影响[J].中医杂志,2015,56(20):1767-1770.
[9]ZHENG Q,GUAN Y,XIA L,et al.Effect of Yi Gong San Decoction on iron homeostasis in a mouse model of acute inflammation[J/OL].Evid Based Complement Alternat Med,2016[2016-04-07].https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4838806.
[10]郑秦,姜一陵,罗梅宏,等.异功散对小鼠慢性病贫血的治疗作用[J].实验动物与比较医学,2017,37(6):1-6.
[11]LASOCKI S,MILLOT S,ANDRIEU V,et al.Phlebotomies or erythropoietin injections allow mobilization of iron stores in a mouse model mimicking intensive care anemia[J].Crit Care Med,2008,36(8):2388-2394.
[12]魏伟,吴希美,李元建,等.药理实验方法学[M].北京:人民卫生出版社,2010:69-72.
[13]张凤奎.慢性疾病性贫血发病机制的研究进展[J].临床血液学杂志,2012,25(11):696-699.
[14]谢坤莹,魏锦.炎症因子在RA合并慢性病贫血发病过程的相关性研究[J].世界最新医学信息文摘,2017,17(40):17-19.
[15]张慧燕.慢性病贫血的发病机制诊断与治疗[J].医学综述,2002,8(4):229-230.
[16]罗梅宏.从脾主运化和运脾生血理论探讨慢性病贫血的中医病机和治疗[J].中医杂志,2013,54(18):1556-1557.
[17]LIANG C,ZHANG S H,CAI Z D,et al.Effects of early intestinal application of sijunzi decoction on immune function in post-operational patients of gastrointestinal tumor[J].Zhongguo Zhong Xi Yi Jie He Za Zhi,2005,25(12):1070-1073.
[18]KIM S J,SHIN H J,LEE B J,et al.The antiinflammatory mechanism of Igongsan in mouse peritoneal macrophages via suppression of nfkappab/caspase-1 activation[J].Phytother Res,2014,28(5):736-744.
[19]LIU X B,NGUYEN N B,MARQUESS K D,et al.Regulation of hepcidin and ferroportin expression by lipopolysaccharide in splenic macrophages[J].Blood Cells Mol Dis,2005,35(1):47-56.
[20]GARDENGHI S,RENAUD T M,MELONI A,et al.Distinct roles for hepcidin and interleukin-6 in the recovery from anemia in mice injected with heat-killed Brucella abortus[J].Blood,2014,123(8):1137-1145.
[21]KIM A,FUNG E,PARIKH S G,et al.A mouse model of anemia of inflammation:complex pathogenesis with partial dependence on hepcidin[J].Blood,2014,123(8):1129-1136.
[22]张引红.补骨脂素对类风湿性关节炎小鼠模型的免疫调节作用[J].中国实验动物学报,2017,25(2):207-210.
[23]张梦思.当归多糖与阿糖胞苷联合注射对人白血病模型小鼠脾脏结构与功能的影响[J].中药药理与临床,2014,30(4):62-66.
[24]ZHONG Z,SANCHEZLOPEZ E,KARIN M,et al.Autophagy,NLRP3inflammasome and auto-inflammatory/immune diseases[J].Clin Exp Rheumatol,2016,34(4 Suppl 98):12-16.
[25]林妘.NF-κB通过清除受损线粒体抑制炎症小体的活化[J].中医肿瘤生物杂志,2016,164(5):896-910.
[26]周露露,牛佳丽,陈陶陶,等.高浓度LPS损伤巨噬RAW264.7线粒体功能的研究[J].吉林医药学院学报,2015,36(1):17-18.