Notch1在新生期小鼠肺炎链球菌感染诱导Th1/Th2失衡

王艺颖; 简鼎; 朱琳; 张瀚之; 李媛媛; 陈诗懿; 张光莉; 罗征秀

doi:10.16016/j.2097-0927.202204021

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Notch1在新生期小鼠肺炎链球菌感染诱导Th1/Th2失衡

基础医学 | 更新时间：2026-01-09

- Notch1在新生期小鼠肺炎链球菌感染诱导Th1/Th2失衡
- Notch1 induces Th1/Th2 imbalance in neonatal mice with Streptococcus pneumoniae infection
- 陆军军医大学学报 2022年44卷第21期页码：2138-2145
- 作者机构：
  
  400014 重庆，重庆医科大学附属儿童医院呼吸科，国家儿童健康与疾病临床医学研究中心，儿童发育与疾病教育部重点实验室，儿科学重庆市重点实验室, 儿童感染免疫重庆市重点实验室
- 作者简介：
  
  [罗征秀，E-mail：luozhengxiu816@163.comLUO Zhengxiu, E-mail: luozhengxiu816@163.com
- 基金信息：
  
  庆市自然科学基金(cstc2020jcyj-msxmX0677);儿童发育疾病研究教育部重点实验室基础研究一般项目(GBRP-202112)
- DOI：10.16016/j.2097-0927.202204021
  中图分类号： R378.12;R392.13;R563.1
- 收稿：2022-04-02，
  
  修回：2022-06-23，
  
  纸质出版：2022-11-15
- 稿件说明：
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王艺颖, 简鼎, 朱琳, 等. Notch1在新生期小鼠肺炎链球菌感染诱导Th1/Th2失衡[J]. 陆军军医大学学报, 2022,44(21):2138-2145.

Yiying WANG, Ding JIAN, Lin ZHU, et al. Notch1 induces Th1/Th2 imbalance in neonatal mice with Streptococcus pneumoniae infection[J]. Journal of Army Medical University, 2022, 44(21): 2138-2145.
王艺颖, 简鼎, 朱琳, 等. Notch1在新生期小鼠肺炎链球菌感染诱导Th1/Th2失衡[J]. 陆军军医大学学报, 2022,44(21):2138-2145. DOI： 10.16016/j.2097-0927.202204021.

Yiying WANG, Ding JIAN, Lin ZHU, et al. Notch1 induces Th1/Th2 imbalance in neonatal mice with Streptococcus pneumoniae infection[J]. Journal of Army Medical University, 2022, 44(21): 2138-2145. DOI： 10.16016/j.2097-0927.202204021.

摘要

目的

研究Notch1信号通路在新生期小鼠肺炎链球菌感染诱导Th1/Th2失衡中的作用。

方法

12只1周龄Balb/c新生小鼠分

为3组(

=4)：新生期肺炎链球菌肺炎(

Streptococcus pneumoniae

pneumonia，S.pp)组、新生期肺炎链球菌肺炎Notch1抑制组[S.pp+DAPT(γ分泌酶抑制剂)

]

和对照组。S.pp组、S.pp+DAPT组小鼠鼻腔滴注10 μL含有1×10

cfu肺炎链球菌的悬液，对照组同期滴注等量PBS。小鼠感染肺炎链球菌后第7天，S.pp+DAPT组鼻腔滴注0.3 mg/kg的Notch1抑制剂DAPT，S.pp组和对照组鼻腔滴注等量PBS。小鼠发育至6周(成年)，检测气道反应性，收集支气管肺泡灌洗液(bronchoalveolar lavage fluid，BALF)行细胞分类计数，肺组织HE染色病理切片观察组织病理变化；Western blot检测肺组织Notch1蛋白水平；RT-PCR检测肺组织

T-bet

、

GATA

3 mRNA水平；流式细胞术检测肺组织Th1、Th2细胞水平并分析Th1/Th2比率。

结果

S.pp+DAPT组Notch1蛋白表达显著低于S.pp组(0.23±0.09

1.94±0.78，

0.01)。S.pp+DAPT组BALF细胞总数、中性粒细胞、单核细胞、淋巴细胞计数均明显低于S.pp组(

0.01)；肺组织中细支气管周围炎症(0.55±0.17

1.52±0.27

0.01)、血管周围炎症(0.05±0.10

0.88±0.25

0.01)、肺间质炎症评分(0.18±0.35

1.57±0.25

0.01)、肺部

GATA

3 mRNA表达(19.11±9.79

177.56±65.95

0.01)、Th2细胞水平[(0.16±0.02)%

(0.28±0.04)%

0.01

]

也显著低于S.pp组；而

T-bet

mRNA表达、Th1细胞水平与S.pp组比较差异无统计学意义(

0.05)，Th1/Th2比值较S.pp组明显增加(14.67±2.60

7.89±0.82

0.01)。S.pp+DAPT组小鼠吸入3.125~50 mg/mL浓度乙酰甲胆碱激发后，气道反应性较S.pp组显著降低(

0.01)。

结论

新生期肺炎链球菌肺炎可能通过激活Notch1信号诱导Th1/Th2失衡，促进气道高反应性及气道炎症形成。

Abstract

Objective

To investigate the role of Notch1 signaling pathway in Th1/Th2 imbalance induced by neonatal

Streptococcus pneumoniae

pneumonia (S.pp) infection.

Methods

Twelve Balb/c neonatal mice (1 week old) were randomly divided into neonatal S.pp group

neonatal S.pp with Notch1 inhibition treatment group (S.pp+DAPT

a γ-secretase inhibitor) and mock-infected control group (control gro

up) (

=4). The S.pp group and the S.pp+DAPT group were infected with 1×10

CFU of

S. pneumoniae

in 10 μL of PBS intranasally

while the control group received equivalent PBS. In 7 d after infection

the S.pp+DAPT groups were given 0.3 mg/kg DAPT by nasal instillation

and the S.pp and the control groups were instilled equivalent PBS. In 5 weeks after infection

airway hyperresponsiveness (AHR) was evaluated by invasive body plethysmography system. Differential cell counts in bronchoalveolar lavage fluid (BALF) were analyzed. HE staining was used to observe the histopathological changes of the lung tissue. Western blotting was used to detect the protein expression of Notch1 protein

and RT-PCR to measure the mRNA levels of

T-bet

and

GATA

3. And the percentages of Th1 and Th2 cells in the lung tissue were determined by flow cytometry and Th1/Th2 ratio was calculated.

Results

Notch1 expression was significantly lower in the S.pp+ DAPT group than the S.pp group (0.23±0.09

1.94±0.78

0.01). The total cell count and counts of neutrophils

monocytes and lymphocytes in the BALF from the S.pp+DAPT group were remarkably lower than those in the S.pp group (

0.01). The S.pp+DAPT mice had obviously lower histologic scores of pulmonary peribronchiolitis

pulmonary perivasculitis and pulmonary alveolitis in lung tissue than the S.pp group (0.55±0.17

1.52±0.27

0.01; 0.05±0.10

0.88±0.25

0.01; 0.18±0.35

1.57±0.25

0.01). The mRNA level of

GATA

3 and percentage of Th2 cells were significantly lower in the S.pp+DAPT group than the S.pp group [19.11±9.79

177.56±65.95

0.01; (0.16±0.02)%

(0.28±0.04)%

0.01

]

. While the mRNA level

T-bet

and percentage of Th1 cells were similar between the 2 groups. The S.pp+DAPT group had significantly higher Th1/Th2 when compared with the S.pp group (14.67±2.60

7.89±0.82

0.01). The AHR in the S.pp+DAPT group mice was significantly decreased when inhaled different concentrations of acetylcholine (3.125~50 mg/mL) as compared with the S.pp group (

0.01).

Conclusion

Neonatal S.pp may induce Th1/Th2 imbalance by activating Notch1 signal

and thus promote airway hyperreactivity and airway inflammation.

关键词

Keywords

references

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