ISSN 1674-3865  CN 21-1569/R
主管：国家卫生健康委员会
主办：中国医师协会
　　　辽宁省基础医学研究所
　　　辽宁中医药大学附属医院
中国中西医结合儿科学

中国中西医结合儿科学 ›› 2017, Vol. 9 ›› Issue (6): 466-471.doi: 10.3969/j.issn.1674-3865.2017.06.002

• 实验论著 • 上一篇    下一篇

清燥救肺汤及其分解剂对肺炎支原体感染小鼠TLR-2/NF-κB信号通路的影响

吴振起，贾晓儒，敏娜，岳志军，王雪峰，张聪聪   

  1. 110032沈阳，辽宁中医药大学附属医院儿科（吴振起，岳志军，王雪峰）；110032沈阳，辽宁中医药大学中医儿科学专业研究生
  • 出版日期:2017-12-25 发布日期:2018-11-19
  • 通讯作者: 吴振起，E-mail：zhenqiwu@163.com
  • 作者简介:吴振起(1974-)，男，医学博士，主任医师，硕士研究生导师。研究方向：中医药防治感染性疾病
  • 基金资助:
    国家自然科学基金面上项目（81373687）；辽宁省科技厅项目（2014020044）

Influence of Qingzao Jiufei decoction and its decomposable agent on TLR-2/NF-κB signal pathway in MP infected mice

WU Zhenqi，JIA Xiaoru，MIN Na，YUE Zhijun，WANG Xuefeng，ZHANG Congcong   

  1. Affiliated Hospital of Liaoning University of TCM,Shenyang 110032，China
  • Online:2017-12-25 Published:2018-11-19

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摘要：
目的
观察清燥救肺汤及其分解剂对肺炎支原体（MP）感染小鼠Toll样受体2（TLR-2）、髓样分化因子88（MyD88)、核因子κB（NF-κB）信号通路的影响，探讨其抗MP的作用机制。
方法
选择SPF级BALB/c小鼠144只，随机分成正常组、模型组、全方组、分解剂Ⅰ组、分解剂Ⅱ组及阿奇霉素组，每组24只。除正常组其他各组制备MP感染模型，造模成功后予药物灌胃治疗，并于感染后第3、7、10、14天进行取材。采用苏木精-伊红染色观察肺脏病理组织学改变、qPCR方法检测肺组织中TLR-2 mRNA变化、ELISA法检测血清中MyD88、TNF-α的含量及Western blot法检测肺组织NF-κB的表达。
结果
MP感染后小鼠肺组织出现间质性炎症改变，7 d时炎症最明显，14 d时炎症逐渐减轻；治疗后各组肺部炎症均有改善，尤以第10、14天全方组、阿奇霉素组明显。MP感染后小鼠肺组织中TLR-2、NF-κB表达水平升高，血清中MyD88、TNF-α含量亦升高（P<0.05），各项指标出现峰值的时间不一致，其中NF-κB出现最早（第3天），TLR-2、TNF-α于第7天达峰值，而MyD88则相对较晚，于第10天达峰值。第7天，全方组即起效（P<0.05），肺组织中TLR-2、NF-κB表达水平下降，血清中MyD88、TNF-α含量亦降低；分解剂Ⅰ组在第10、14天亦能发挥类似全方组的作用，但较全方组偏弱（P>0.05）；分解剂Ⅱ组在第14天TLR-2 mRNA表达略升高。
结论
清燥救肺汤抗MP感染的机制与减少促炎因子的释放，调控TLR-2/NF-κB信号通路传导有关，其中分解剂Ⅰ起主要作用。

关键词: 肺炎支原体, 清燥救肺汤, TLR-2/NF-κB信号通路, 小鼠

Abstract:
Objective
To observe the effect of Qingzao Jiufei decoction (QJD) and its decomposable agent on the toll-like receptors(TLR) 2/myeloid differentiation primary response gene 88 (MyD88) / (NF-κB) signal pathway in MP infected mice，and to explore its mechanism of action of resistance to MP.
Methods
Totally 144 BALB/c mice, about 20 g in weight, were randomly divided into normal group(A),model group(B),QJD group(C),QJD decomposable agentⅠgroup(D),QJD decomposable agent Ⅱ group(E) and azithromycin group(F),with 24 mice in each group. Each treatment group was made into MP infection model and the corresponding group was given corresponding drugs by intragastric administration after modeling. The serum and the lung tissues were detected after 3,7,10 and 14 days of MP infection. HE staining was used to observe the pathological change of lung tissues, qPCR was used to detect the changes of TLR-2 mRNA in lung tissues, ELISA was used to detect the expressions of MyD88 and TNF-α in serum, and Western blot was used to detect the expression levels of NF-κB in lung tissues.
Results
The interstitial inflammatory changes of lung occurred 3d after MP infection，which were most significant on the 7d，and then gradually became less and less significant. Pulmonary inflammation in each treatment group was improved, especially in Group C and Group F on the 10d and 14d. The expression level of TLR-2 and NF-κB in the lung tissue of mice after MP infection increased, and the content of MyD88 and TNF-α in serum was also increased(P<0.05).The peak time of each indicator was different with NF-κB appearing earliest (3rd day),TLR-2 and TNF-α peaking on the 7th day, while MyD88 was relatively late，peaking on the 10th day. On the 7th day in Group C(P＜0.05),the expression level of TLR-2 and NF-κB in lung tissue decreased, and the content of MyD88 and TNF-α in serum was also decreased. Group D also played a similar role to Group C on the 10th day and 14th days, but weaker than Group C (P＞0.05); in Group E on the 14th day TLR-2 mRNA expression slightly increased.
Conclusion
The mechanism of QJD against MP infection is associated with reducing the release of proinflammatory factors and regulating the conduction of TLR-2/NF-κB signaling pathway，in which the decomposition agent Ⅰ plays a major role.

Key words: Mycoplasma pneumoniae, Qingzao Jiufei decoction, TLR-2/NF-κB signal pathway, Mice