摘要
目的:建立大肠埃希菌(Escherichia coli)感染哺乳期西藏小型猪腹泻模型,研究葛根芩连汤(GQT)治疗婴幼儿E.coli感染性腹泻作用机制。方法:按照体重灌胃不同剂量的E.coli,灌胃后每3 h进行体重,体温,和腹泻状况的观察并收集粪便进行E.coli计数;分别设置阳性药蒙脱石散组(93.5 mg·kg~(-1)),GQT高、中、低剂量组(含生药12.1,6.05,1.21 g·kg~(-1)),给药3 d,每天2次。每次给药后观察腹泻指数、体重变化,统计生存率,检测粪便中E.coli菌落数,血液白细胞数(WBC),酶联免疫吸附法(ELISA)检测血清肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)含量。结果:E.coli剂量为1×109CFU·kg~(-1)时,6 h仍为水样泻,粪便E.coli计数>1×1011CFU·g-1,无明显自愈倾向;GQT低剂量组可显著抑制腹泻,中、高剂量组无明显止泻作用;高、中、低剂量组可显著降低粪便E.coli和血液WBC的数量,抑制血清TNF-α和IL-6的表达。结论:当E.coli灌服剂量为1×109CFU·kg~(-1)时,6 h仍为水样泻,且粪便E.coli计数>1×1011CFU·g-1,则判定造模成功;GQT可通过抗炎和抑制E.coli的增殖减轻E.coli感染哺乳期小型猪腹泻。其抗炎作用可能与减少炎症因子TNF-α和IL-6有关。
Objective: To establish the Escherichia coli-infected diarrhea model of suckling Tibetan minipigs,in order to study the mechanism of Gegen Qinlian Tang(GQT) on E. coli-infected diarrhea in infants.Method: The mini-pig diarrhea model was established through oral administration with pathogenic E. coli. The animals were divided into control group(Montmorillonite powder,93. 5 mg·kg~(-1)),low-dose group(1. 21 g·kg~(-1)),middle-dose group(6. 05 g·kg~(-1)) and high-dose group(12. 1 g·kg~(-1)). Fecal E. coli count and white blood cell count were detected after each administration,and the serum tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) were detected by enzyme linked immunosorbent assay(ELISA) kit. The diarrhea index,weight change and survival rate were observed. Result: When the dose of E. coli was 1 × 109 CFU·kg~(-1),the watery diarrhea was maintained at 6 h,the count of fecal coliform was higher than 1 × 1011 CFU·g-1,and there was no obvious trend of self-cure. High,medium and low-dose GQT groups showed significant reduction in fecal E. coli count and blood white blood count,and inhibition of the expressions of serum TNF-α and IL-6. Low-dose group showed significant inhibition of diarrhea,and middle and high-dose groups had no obvious antidiarrheal effect.Conclusion: When the dose of E. coli was 1 × 109 CFU·kg~(-1),the feces were still watery at 6 h,and the E. coli count of fecal was higher than 1 × 1011 CFU·g-1,which proved that the E. coli-infected diarrhea mini-pig model was successfully established. GQT could relieve E. coli-infected diarrhea among suckling pigs by resisting inflammation and inhibiting the proliferation of E. coli. The anti-inflammatory effect may be related to the reduction of inflammatory factors TNF-α and IL-6.
引文
[1]王君梅.中药汤剂清金化痰汤加减治疗小儿支原体肺炎临床疗效观察[J].海峡药学,2017,29(4):196-197.
[2]孙谦,路娜娜,孙雯,等.探究儿童抗生素相关性腹泻中药学干预的作用与价值[J].中外医疗,2017,36(8):1-3.
[3]崔新洁,夏瑾,邵铁娟,等.中药抗耐甲氧西林金黄色葡萄球菌生物膜研究进展[J].中国中医药信息杂志,2017,24(2):132-136.
[4]王新华.抗生素联合中药保留灌肠治疗慢性盆腔炎疗效观察[J].临床合理用药杂志,2017,10(1):48-49.
[5]张振巍,张娜娜,石磊,等.中药“抗生素”临床应用思考[J].中医学报,2014,29(7):1019-1020.
[6]刘惠瑾,黄春霞,牛艳慧.中西药结合治疗婴幼儿支气管肺炎抗生素相关性腹泻的临床疗效观察[J].中成药,2017,39(5):1103-1105.
[7]张凌波,郝瑞芳.葛根芩连汤合平胃散加减治疗小儿秋季腹泻(湿热型)40例临床研究[J].亚太传统医药,2016,12(20):126-127.
[8]叶晶晶,钟夏君.葛根芩连汤加减治疗小儿秋季腹泻的疗效观察[J].海峡药学,2015,27(8):176-177.
[9]阿米娜·克然木.中西医结合治疗婴儿腹泻疗效观察[J].世界最新医学信息文摘,2015,15(18):142.
[10]彭佩佩.加味葛根芩连汤治疗小儿湿热型泄泻的临床观察[D].石家庄:河北医科大学,2012.
[11]张海英.葛根芩连汤保留灌肠治疗小儿秋季腹泻的临床观察[C]∥中华中医药学会.第四次全国民间传统诊疗技术与验方整理研究学术会论文集.北京,2011:2.
[12]曾菊香.葛根芩连汤加味治疗婴幼儿腹泻60例[J].现代中西医结合杂志,2011,20(17):2156-2157.
[13]赵益,赖小东,叶争荣,等.葛根芩连汤对溃疡性结肠炎模型大鼠抗氧化及抗炎的作用机制[J].中华中医药杂志,2016,31(5):1741-1745.
[14]毛莹,张贵君,彭慧,等.葛根芩连汤药效组分解热抗炎药效学研究[J].辽宁中医药大学学报,2014,16(1):30-32.
[15]杨婧,杨丽珍,王亚贤,等.不同制备工艺葛根芩连汤抗炎抑菌作用比较[J].中国实验方剂学杂志,2013,19(1):224-226.
[16]郭震浪,苏振宁,戴韵峰,等.葛根芩连汤治疗小儿轮状病毒性肠炎疗效的Meta分析[J].中国实验方剂学杂志,2015,21(24):199-203.
[17]陈丽红,唐于平,王强.不同提取方法制备的葛根芩连配方颗粒抑菌药效研究[J].中成药,2012,34(10):2016-2017.
[18]王新,杨婧,马凯,等.葛根芩连汤防治小鼠感染大肠埃希菌的作用观察[J].黑龙江畜牧兽医,2017,(14):147-149,155.
[19]刘茜明,杨光勇,何光志,等.葛根芩连汤对抗生素相关腹泻肠道乳酸杆菌属的影响研究[J].家畜生态学报,2016,37(11):74-78.
[20]刘茜明,杨光勇,何光志,等.FQ-PCR技术分析葛根芩连汤对抗生素相关性腹泻模型肠道双歧杆菌属的影响[J].时珍国医国药,2016,27(8):1820-1823.
[21]Yandza T,Tauc M,Saint-Paul M,et al.The pig as a preclinical model for intestinal ischemia-reperfusion and transplantation studies[J].J Surg Res,2012,178(2):807-819.
[22]王俊丽,张要齐,孙雪峰,等.中药复方对猪大肠埃希菌的体外抑菌活性研究[J].中兽医学杂志,2013(1):6-8.
[23]陈甜甜,杨忠福,刘建柱.猪大肠杆菌病研究进展[J].猪业科学,2008(8):20-25.
[24]李晋涛,魏静,牟芝蓉,等.人源轮状病毒的小型猪腹泻模型建立[J].中国比较医学杂志,2006,16(9):577.
[25]崔立.实验用小型猪种群净化技术及其不同模型的建立和应用[D].南京:南京农业大学,2005.
[26]刘宇政,章军,王跃生,等.葛根芩连汤剂量相关问题探讨[J].中国实验方剂学杂志,2010,16(16):216-218.
[27]罗佳波,谭晓梅,余林中,等.葛根芩连汤配伍规律的研究[J].中草药,2005,36(4):512-518.
[28]张赛奇.葛芪复方对仔猪大肠杆菌性腹泻作用效果研究[D].北京:中国农业科学院,2015.
[29]邱爱珠,陈宝田.葛根芩连汤证为感染性腹泻病治的论证研究[J].热带医学杂志,2010,10(6):640-642.
[30]姜丽霞.细菌和病毒感染性腹泻患儿IL-1、IL-6、IL-8和TNF-α水平观察[J].检验医学,2006,21(1):88..
[31]兰洁.降钙素原、白介素-6、C反应蛋白鉴别感染性腹泻的灵敏度、特异性分析[J].现代仪器与医疗,2017,23(3):87-89.
[32]杨立华,傅晓凤,聂微萱,等.感染性腹泻患儿炎前细胞因子检测及临床意义[J].中国实用儿科杂志,2008,23(9):697-698.
[33]高萍芝,王殿敏,王国利,等.感染性腹泻息儿检测血清白细胞介素-6的临床意义[J].临床儿科杂志,1998,16(4):237-238.
[34]邱赛红,孙必强,李磊,等.常用苦寒药过量使用对胃黏膜屏障功能影响的实验研究[J].中国医药导刊,2007,9(2):140-142.
[35]邱赛红,李飞艳,尹健康,等.9味苦寒药对小鼠胃肠运动与肝肾功能影响的实验研究[J].湖南中医学院学报,2004,24(5):1-3,6.
[36]朱朝敏,幸琳琳.感染性腹泻的发病机制[J].实用儿科临床杂志,2010,25(19):1456-1457.
