清肺透邪方对MPP小鼠炎症因子IL-10、IL-17疗效机制研究
|
摘要
目的:通过肺炎支原体滴鼻方法建立感染幼龄Balb/c小鼠MPP模型,采用Elisa法检测小鼠外周血中IL-17、IL-10表达水平变化,观察清肺透邪方对肺炎支原体肺炎(MPP)小鼠治疗效果及其时效关系,为临床治疗肺炎支原体肺炎提供实验依据。方法:90只幼龄Balb/c小鼠随机分为正常组、模型组、阿奇霉素组、中药组和中药+阿奇组,共5组,每组18只,除正常组外,其余4组均建立MPP模型。连续滴鼻3 d后,阿奇霉素组小鼠予阿奇霉素0.2 mL/(d·20 g)灌胃,中药组予0.2 mL/(d·20 g)灌胃,中药+阿奇组予分别阿奇霉素和清肺透邪方颗粒剂各0.2 mL/(d·20 g)灌胃,正常组与模型组予等体积生理盐水灌胃。分别于给药后第3、7、14天处死取材,每组6只采用Elisa法检测外周血中IL-10、IL-17表达水平变化,观察清肺透邪方对MPP小鼠的治疗作用及其时效关系。结果:感染第3、7天时,5组间IL-10水平比较均无统计学差异;感染第14天时模型组IL-10水平均低于正常组(P<0.05),3组给药组均明显高于模型组(P<0.05),且中药+阿奇组明显高于其他两组给药组(P<0.05)。感染第3天时,模型组、阿奇霉素组、中药组、中药+阿奇组小鼠IL-17细胞水平含量均高于正常组(P<0.05),3组给药组均低于模型组(P<0.05),且给药组组间比较无统计学差异;感染第7、14天时模型组与3组给药组小鼠IL-17水平均高于正常组(P<0.05),且中药+阿奇组明显低于其他两组给药组(P<0.05)。结论:清肺透邪方可以有效调控MPP小鼠IL-10、IL-17因子的表达,且联用阿奇霉素效果更优,这可能是清肺透邪方防治MPP以及减轻其炎症反应的作用机制之一。
Objective :To establish a model of MPP in Balb/c mice infected with mycoplasma pneumoniae by nose dropping. The expression of IL-10 and IL-17 in peripheral blood of mice was detected by Elisa method. The therapeutic effect of Qingfei Touxie Decoction on mycoplasma pneumoniae pneumonia in mice and its time-effect relationship were observed,so as to provide experimental basis for clinical treatment of mycoplasma pneumoniae pneumonia. Methods :Ninety young Balb/c mice were randomly divided into five groups :normal group,model group,azithromycin group,traditional Chinese medicine group and traditional Chinese medicine+azithromycin group. Eighteen mice in each group were divided into five groups. MPP models were established in all four groups except the normal group. After three consecutive days of nasal drip,mice in azithromycin group were given 0.2 mL/(d·20 g)intragastric administration of azithromycin,0.2 mL/(d·20 g)intragastric administration of traditional Chinese medicine group,0.2 mL/(d·20 g)intragastric administration of azithromycin and Qingfei Tuixie Decoction granules respectively,and 0.2 mL/(d·20 g)intragastric administration of normal saline and model group. Samples were sacrificed on the 3 rd,7 th and 14 th day after administration. The expression levels of IL-17 and IL-10 in peripheral blood were detected by Elisa method. The therapeutic effect of Qingfei Touxie Decoction on MPP mice and its time-effect relationship were observed. Results :At the 3 rd and 7 th day of infection,there was no significant difference among the five groups in the levels of IL-10. On the 14 th day of infection,the IL-10 level in the model group was lower than that in the normal group(P<0.05),and the IL-10 level in the three drug groups of drug administration was significantly higher than that in the model group(P<0.05),and that in the Chinese medicine + azithromycin group was significantly higher than that in the other two groups of drug administration(P<0.05). On the 3 rd day of infection,the levels of IL-17 cells in model group,azithromycin group,traditional Chinese medicine group and traditional Chinese medicine + azithromycin group were higher than those in normal group(P<0.05),and the levels of IL-17 in three groups of drug administration were lower than those in model group(P<0.05). There was no statistical difference between the three groups. On the 7 th and 14 th day of infection,the levels of IL-17 in model group and three groups of drug administration were higher than those in normal group(P<0.05),and the traditional Chinese medicine + azithromycin group was significantly lower than the other two groups of drug administration(P<0.05). Conclusion :Qingfei Touxie Decoction can effectively regulate the expression of IL-10 and IL-17 in MPP mice,and the combination of azithromycin is more effective,which may be one of the mechanisms of Qingfei Touxie Decoction in preventing MPP and reducing its inflammatory reaction.
Objective :To establish a model of MPP in Balb/c mice infected with mycoplasma pneumoniae by nose dropping. The expression of IL-10 and IL-17 in peripheral blood of mice was detected by Elisa method. The therapeutic effect of Qingfei Touxie Decoction on mycoplasma pneumoniae pneumonia in mice and its time-effect relationship were observed,so as to provide experimental basis for clinical treatment of mycoplasma pneumoniae pneumonia. Methods :Ninety young Balb/c mice were randomly divided into five groups :normal group,model group,azithromycin group,traditional Chinese medicine group and traditional Chinese medicine+azithromycin group. Eighteen mice in each group were divided into five groups. MPP models were established in all four groups except the normal group. After three consecutive days of nasal drip,mice in azithromycin group were given 0.2 mL/(d·20 g)intragastric administration of azithromycin,0.2 mL/(d·20 g)intragastric administration of traditional Chinese medicine group,0.2 mL/(d·20 g)intragastric administration of azithromycin and Qingfei Tuixie Decoction granules respectively,and 0.2 mL/(d·20 g)intragastric administration of normal saline and model group. Samples were sacrificed on the 3 rd,7 th and 14 th day after administration. The expression levels of IL-17 and IL-10 in peripheral blood were detected by Elisa method. The therapeutic effect of Qingfei Touxie Decoction on MPP mice and its time-effect relationship were observed. Results :At the 3 rd and 7 th day of infection,there was no significant difference among the five groups in the levels of IL-10. On the 14 th day of infection,the IL-10 level in the model group was lower than that in the normal group(P<0.05),and the IL-10 level in the three drug groups of drug administration was significantly higher than that in the model group(P<0.05),and that in the Chinese medicine + azithromycin group was significantly higher than that in the other two groups of drug administration(P<0.05). On the 3 rd day of infection,the levels of IL-17 cells in model group,azithromycin group,traditional Chinese medicine group and traditional Chinese medicine + azithromycin group were higher than those in normal group(P<0.05),and the levels of IL-17 in three groups of drug administration were lower than those in model group(P<0.05). There was no statistical difference between the three groups. On the 7 th and 14 th day of infection,the levels of IL-17 in model group and three groups of drug administration were higher than those in normal group(P<0.05),and the traditional Chinese medicine + azithromycin group was significantly lower than the other two groups of drug administration(P<0.05). Conclusion :Qingfei Touxie Decoction can effectively regulate the expression of IL-10 and IL-17 in MPP mice,and the combination of azithromycin is more effective,which may be one of the mechanisms of Qingfei Touxie Decoction in preventing MPP and reducing its inflammatory reaction.
引文
[1] Stenger S,Hanson DA,Teitelbaum R,et al. An antimicrobial activity of citolytic T cells mediated by granulysin[J]. Science,1998,282(5386):121-125.
[2] Adam N Odeh,Jerry W Simecka. Regulatory CD4+CD25+T cells dampen inflammatory disease in murine mycoplasma pneumonia and promote IL-17 and IFN-γresponses[J].PLoS One,2016,11(5):e0155648.
[3]孙诗炜,赵长江,宋锦萍,等. 4246例儿童肺炎支原体感染流行特征[J].安徽医学,2016,37(11):1374-1378.
[4] Atkinson T P,Balish M F,Waites K B. Epidemiology,clinical manifestations,pathogenesis and laboratory detection of Mycoplasma pneumoniae infections[J]. FEMS Microbiol Rev,2008,32(6):956-973.
[5]陈正荣,严永东.小儿肺炎支原体感染流行病学特征[J].中国实用儿科杂志,2015,30(3):180-183.
[6]孙汉庆.2006~2010年肺炎支原体肺炎发病情况调查分析[J].安徽医药,2012,16(2)222-224.
[7] ZHAO F,LIU G,WU J,et al. Surveillance of macrolide-resistant mycoplasma pneumoniae in Beijing,China,from 2008 to 2012[J].Antimicrob Agents Chemother,2013,57(3):1521-1523.
[8]毕美芬,沈立飚,黄如红,等.加味玉屏风散防治支原体肺炎后反复呼吸道感染[J].浙江中医杂志,2012,47(8):563.
[9]曹文才.中药佐治肺炎支原体肺炎38例临床观察[J].中医临床研究,2012,4(4):100.
[10]何军,张姝,许琦,等.肺炎支原体肺炎患儿血清中IL-9、IL-17水平[J].贵阳医学院学报,2015,40(7),712-714.
[11]蔡建敏,陈同辛,王振海,等.肺炎支原体肺炎患儿血清白细胞介素-10、转化生长因子-β1检测的意义[J].实用儿科临床杂志,2007,22(4):256.
[12]潘黎明,王跃,张海英.IL-10在小儿支原体肺炎发病机制中的作用[J].中国实验诊断学,2009,13(3):376-378.
[13]尹嘉宁. CysLTs、IL-10、TNF-α在小儿支原体肺炎发病机制中的作用[D].长春:吉林大学,2008.
[14]刘芳,赵宇华,陈霜慧.IL-6、IL-8及IL-10在肺炎支原体肺炎患儿血清中的表达及临床意义[J].实用临床医药杂志,2015,19(17):61-63,66.
[15]熊远青,黄河,姚霖,等.支原体肺炎患儿外周血Treg/Th17细胞平衡变化及临床意义[J].中国医学创新,2015,12(36):59-61.
[16]杜许芳,周炯英. Th17/Treg失衡在肺炎支原体肺炎患儿中的作用及其机制[J].临床与病理杂志,2016,36(12):2017-2021.
[17]韦媛媛,徐峰,陈侠,等.桑白皮总黄酮的镇咳祛痰作用[J].沈阳药科大学学报,2009,26(8):644-647.
[18]李倩楠,葛晓群.黄芩苷的解热作用及对细胞因子的影响[J].中国中药杂志,2010,35(8):1068-1072.
[19]付璟,石继和.黄芩素体外抑菌与体内抗炎作用研究[J].中国药房,2014,25(23):2136-2138.
[20]孙艳平,焦晓黎,吴秉纯.8味中药提取物体外抗肺炎支原体的试验研究[J].陕西中医,2008,29(6):727.
[21]杨昕宇,肖长芳,张凯熠,等.麻黄临床应用与药理作用研究进展[J].中华中医药学刊,2015,33(12):2874-2877.
[22]董文婷.麻黄-石膏药对配伍的化学成分、药效及代谢组学研究[J].齐齐哈尔医学院学报,2016,37(33):4191-4193.
[23] KOU JP,SUN Y,LIN YW,et al. Anti-inflammatory activities of aqueous extract from radix Ophiopogon japonicas and its two constituents[J].Biol Pharm Bull,2005,28(7):1234-1238.
[24]张海防,窦昌贵,刘晓华,等.虎杖提取物抗炎作用的实验研究[J].药学进展,2003,27(4):230-233.
[25]王志洁.虎杖大黄素抗HSV-2,CVB3病毒的作用初探[J].安徽中医药学院学报,1999,18(3):42-44.
[26]王永奇,邢福有,刘凡亮,等.紫苏子镇咳、祛痰、平喘作用的药理研究[J].中南药学,2003,1(3):136-138.
[27]肖朝霞,蒋萌蒙,王向军.杏仁的功能性及其药理研究进展[J].试验研究,2011(11):71-73.
[28]张树臣,杨晓静.白花桔梗皂苷与紫花桔梗的药理作用的比较[J].中成药研究,1984(2):37.
[2] Adam N Odeh,Jerry W Simecka. Regulatory CD4+CD25+T cells dampen inflammatory disease in murine mycoplasma pneumonia and promote IL-17 and IFN-γresponses[J].PLoS One,2016,11(5):e0155648.
[3]孙诗炜,赵长江,宋锦萍,等. 4246例儿童肺炎支原体感染流行特征[J].安徽医学,2016,37(11):1374-1378.
[4] Atkinson T P,Balish M F,Waites K B. Epidemiology,clinical manifestations,pathogenesis and laboratory detection of Mycoplasma pneumoniae infections[J]. FEMS Microbiol Rev,2008,32(6):956-973.
[5]陈正荣,严永东.小儿肺炎支原体感染流行病学特征[J].中国实用儿科杂志,2015,30(3):180-183.
[6]孙汉庆.2006~2010年肺炎支原体肺炎发病情况调查分析[J].安徽医药,2012,16(2)222-224.
[7] ZHAO F,LIU G,WU J,et al. Surveillance of macrolide-resistant mycoplasma pneumoniae in Beijing,China,from 2008 to 2012[J].Antimicrob Agents Chemother,2013,57(3):1521-1523.
[8]毕美芬,沈立飚,黄如红,等.加味玉屏风散防治支原体肺炎后反复呼吸道感染[J].浙江中医杂志,2012,47(8):563.
[9]曹文才.中药佐治肺炎支原体肺炎38例临床观察[J].中医临床研究,2012,4(4):100.
[10]何军,张姝,许琦,等.肺炎支原体肺炎患儿血清中IL-9、IL-17水平[J].贵阳医学院学报,2015,40(7),712-714.
[11]蔡建敏,陈同辛,王振海,等.肺炎支原体肺炎患儿血清白细胞介素-10、转化生长因子-β1检测的意义[J].实用儿科临床杂志,2007,22(4):256.
[12]潘黎明,王跃,张海英.IL-10在小儿支原体肺炎发病机制中的作用[J].中国实验诊断学,2009,13(3):376-378.
[13]尹嘉宁. CysLTs、IL-10、TNF-α在小儿支原体肺炎发病机制中的作用[D].长春:吉林大学,2008.
[14]刘芳,赵宇华,陈霜慧.IL-6、IL-8及IL-10在肺炎支原体肺炎患儿血清中的表达及临床意义[J].实用临床医药杂志,2015,19(17):61-63,66.
[15]熊远青,黄河,姚霖,等.支原体肺炎患儿外周血Treg/Th17细胞平衡变化及临床意义[J].中国医学创新,2015,12(36):59-61.
[16]杜许芳,周炯英. Th17/Treg失衡在肺炎支原体肺炎患儿中的作用及其机制[J].临床与病理杂志,2016,36(12):2017-2021.
[17]韦媛媛,徐峰,陈侠,等.桑白皮总黄酮的镇咳祛痰作用[J].沈阳药科大学学报,2009,26(8):644-647.
[18]李倩楠,葛晓群.黄芩苷的解热作用及对细胞因子的影响[J].中国中药杂志,2010,35(8):1068-1072.
[19]付璟,石继和.黄芩素体外抑菌与体内抗炎作用研究[J].中国药房,2014,25(23):2136-2138.
[20]孙艳平,焦晓黎,吴秉纯.8味中药提取物体外抗肺炎支原体的试验研究[J].陕西中医,2008,29(6):727.
[21]杨昕宇,肖长芳,张凯熠,等.麻黄临床应用与药理作用研究进展[J].中华中医药学刊,2015,33(12):2874-2877.
[22]董文婷.麻黄-石膏药对配伍的化学成分、药效及代谢组学研究[J].齐齐哈尔医学院学报,2016,37(33):4191-4193.
[23] KOU JP,SUN Y,LIN YW,et al. Anti-inflammatory activities of aqueous extract from radix Ophiopogon japonicas and its two constituents[J].Biol Pharm Bull,2005,28(7):1234-1238.
[24]张海防,窦昌贵,刘晓华,等.虎杖提取物抗炎作用的实验研究[J].药学进展,2003,27(4):230-233.
[25]王志洁.虎杖大黄素抗HSV-2,CVB3病毒的作用初探[J].安徽中医药学院学报,1999,18(3):42-44.
[26]王永奇,邢福有,刘凡亮,等.紫苏子镇咳、祛痰、平喘作用的药理研究[J].中南药学,2003,1(3):136-138.
[27]肖朝霞,蒋萌蒙,王向军.杏仁的功能性及其药理研究进展[J].试验研究,2011(11):71-73.
[28]张树臣,杨晓静.白花桔梗皂苷与紫花桔梗的药理作用的比较[J].中成药研究,1984(2):37.