Experimental study of <italic style="font-style: italic">Xiao</italic>’<italic style="font-style: italic">er Jianpi Yifei</italic> Formula improving airway hyperresponsiveness in asthmatic mice

CHEN Xun; YAN Xiaoru; LI Min; WAN Tong; JIANG Yanwen; JI Xiaohua

doi:10.3969/j.issn.1006-2157.2023.09.012

您当前的位置：

首页 >

文章列表页 >

Experimental study of Xiao’er Jianpi Yifei Formula improving airway hyperresponsiveness in asthmatic mice

Experimental Studies | 更新时间：2023-10-09

- Experimental study of Xiao’er Jianpi Yifei Formula improving airway hyperresponsiveness in asthmatic mice
- Journal of Beijing University of Traditional Chinese Medicine Vol. 46, Issue 9, Pages: 1267-1279(2023)
- 作者机构：
  
  中国中医科学院西苑医院　北京　100091
- 作者简介：
  
  Prof. JI XiaoHua, Chief physician. Xiyuan Hospital, China Academy of Chinese Medical Sciences, No.1, Xiyuan Playground, Haidian District, Beijing 100091. E-mail: 13611199408@163.com
- 基金信息：
  
  National Natural Science Foundation of China(82104934)
- DOI：10.3969/j.issn.1006-2157.2023.09.012
  CLC： R285.5
- Received：29 March 2023，
  
  Online First：27 July 2023，
  
  Published：30 September 2023
- 稿件说明：
移动端阅览
CHEN Xun, YAN Xiaoru, LI Min, et al. Experimental study of Xiao’er Jianpi Yifei Formula improving airway hyperresponsiveness in asthmatic mice[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023, 46(9): 1267-1279. DOI: 10.3969/j.issn.1006-2157.2023.09.012.
DOI：

CHEN Xun, YAN Xiaoru, LI Min, et al. Experimental study of Xiao’er Jianpi Yifei Formula improving airway hyperresponsiveness in asthmatic mice[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023, 46(9): 1267-1279. DOI: 10.3969/j.issn.1006-2157.2023.09.012. DOI：

摘要

目的

探讨小儿健脾益肺方通过高迁移率组蛋白B1(HMGB1)/Toll样受体4(TLR4)/核转录因子-κB(NF-κB)炎症信号通路改善哮喘小鼠气道高反应的作用机制。

方法

60只SPF级Balb/c小鼠随机数字表法分为空白组、模型组、布地奈德组、小儿健脾益肺方组、布地奈德＋小儿健脾益肺方组，用卵清白蛋白(OVA)致敏、激发建立小鼠急性哮喘模型，分别予布地奈德(0.2 g/L)、小儿健脾益肺方(1.875 g/kg)干预。肺功能检测气道阻力，支气管肺泡灌洗液吉姆萨染色检测小鼠各种细胞占比情况，肺组织切片HE及PAS染色观察，ELISA检测血清及支气管肺泡灌洗液中OVA-IgE、白细胞介素(IL)-6、IL-1β、γ干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)含量，蛋白质印迹法及免疫荧光检测HMGB1、髓样分化因子88(MyD88)、TLR4、p-NF-κB/NF-κB等通路蛋白表达情况。

结果

与空白组比较，模型组气道阻力明显升高(

0.01)，支气管肺泡灌洗液中嗜酸性粒细胞比例明显升高(

0.01)，组织病理学染色较空白组出现明显炎性细胞浸润、杯状细胞增生、气道上皮水肿、管腔狭窄，血清及支气管肺泡灌洗液中OVA-IgE、IL-6、IL-1β、IFN-γ、TNF-α表达升高(

0.01)，HMGB1、MyD88、TLR4、NF-κB等通路蛋白表达升高(

0.05)，NF-κB磷酸化程度升高(

0.05)。与模型组比较，布地奈德组、小儿健脾益肺方组、布地奈德＋小儿健脾益肺方组气道阻力降低，支气管肺泡灌洗液中巨噬细胞占比升高、嗜酸性粒细胞降低(

0.05)，组织病理学染色结果显示炎症细胞浸润改善、平滑肌层增厚缓解、上皮完整性增加，杯状细胞化生减少，HE评分、PAS评分与模型组比较差异具有统计学意义(

0.05)。血清和支气管肺泡灌洗液中，小儿健脾益肺方组、布地奈德组、布地奈德＋小儿健脾益肺方组与模型组比较均能明显降低各项指标的含量(

0.01)；与布地奈德组比较，小儿健脾益肺方组血清OVA-IgE含量明显升高(

0.05)，小儿健脾益肺方组和布地奈德＋小儿健脾益肺方组HMGB1含量明显降低(

0.05)。蛋白质印迹法检测结果显示，与模型组比较，小儿健脾益肺方组、布地奈德组、布地奈德＋小儿健脾益肺方组均能有效降低HMGB1、MyD88和TLR4蛋白的表达和NF-κB的磷酸化程度，差异具有统计学意义(

0.01)；与布地奈德组相比，小儿健脾益肺方组HMGB1的表达明显下降，而TLR4、MyD88的表达升高，布地奈德＋小儿健脾益肺方组HMGB1、TLR4、MyD88的表达均明显下降，差异均具有统计学意义(

0.05)。免疫荧光检测结果显示，小儿健脾益肺方组、布地奈德组、布地奈德＋小儿健脾益肺方组HMGB1与模型组比较明显降低(

0.01)，布地奈德＋小儿健脾益肺方组的HMGB1和TLR4较布地奈德组均明显降低(

0.05)。

结论

小儿健脾益肺方和布地奈德均能有效缓解哮喘相关病理改变，降低炎症因子表达，抑制HMGB1/TLR4/NF-κB炎症信号通路，改善气道高反应。小儿健脾益肺方和布地奈德联合应用优于布地奈德单独应用。HMGB1/TLR4/NF-κB炎症信号通路是健脾益肺方改善儿童哮喘气道高反应性的潜在作用机制。

Abstract

Objective

We sought to explore the mechanism of

Xiao

’

er Jianpi Yifei

Formula in improving airway hyperresponsiveness in asthmatic mice signaling pathway through the HMGB1/TLR4/NF-κB inflammatory signaling pathway.

Methods

Sixty SPF Balb/c mice were randomly divided into the control group

the model group

the budesonide group

the

Xiao

’

er Jianpi Yifei

Formula group

and the budesonide +

Xiao

’

er Jianpi Yifei

Formula group. The acute asthma model was established by sensitization and stimulation with ovalbumin(OVA). Budesonide(0.2 g/L) and

Xiao

’

er Jianpi Yifei

Formula(1.875 g/kg) were used for intervention. The degree of airway hyperresponsiveness was evaluated by measuring airway resistance. The proportion of different types of cells in bronchoalveolar lavage fluid(BALF) was detected by Giemsa staining. Pathological changes were observed by HE staining and PAS staining in lung tissue sections. The expression levels of OVA-IgE

IL-6

IL-1β

IFN-γ

and TNF-α in serum and BALF were evaluated by ELISA

and the expression of HMGB1

MyD88

TLR4

and p-NF-κB/NF-κB were detected by Western blotting and immunofluorescence.

Results

Compared to the control group

the model group showed a significant increase in airway resistance(

0.01)

an obvious increase in the proportion of eosinophils in bronchoalveolar lavage fluid(

0.01)

significant inflammatory cell infiltration

goblet cell hyperplasia

epithelial swelling

luminal stenosis in tissue pathology staining

and a significant increase in the expression levels of OVA-IgE

IL-6

IL-1β

IFN-γ

and TNF-α in serum and BALF(

0.01). The expression levels of HMGB1

MyD88

TLR4

and NF-κB were also elevated(

0.05). The phosphorylation of NF-κB was increased(

0.05). Compared to the model group

the budesonide group

the

Xiao

’

er Jianpi Yifei

Formula group

and the budesonide+

Xiao

’

er Jianpi Yifei

Formula group exhibited reduced airway resistance

an increased proportion of macrophages in BALF

and reduced eosinophil counts(

0.05). The histopathological staining result showed that the infiltration of inflammatory cells was improved

the smooth muscle layer thickening reduced

the epithelial integrity was increased

and the metaplasia of Goblet cell was reduced. HE scores and PAS scores compared to the model group were statistical significant(

0.05). In the serum and BALF

the contents of various indexes in the

Xiao

’

er Jianpi Yifei

Formula group

the budesonide group

and the budesonide+

Xiao

’

er Jianpi Yifei

Formula group were lower than those in the model group(

0.01); Compared to the budesonide group

the serum OVA-IgE content in the

Xiao

’

er Jianpi Yifei

Formula group was greater (

0.05)

while the HMGB1 content in the

Xiao

’

er Jianpi Yifei

Formula group and the budesonide+

Xiao

’

er Jianpi Yifei

Formula group was lower(

0.05). Western blotting result showed that

compared to the model group

the expression of HMGB1

MyD88

and TLR4 proteins

and the phosphorylation degree of NF-κB could be effectively reduced (

0.01) in the

Xiao

’

er Jianpi Yifei

Formula group

the budesonide group

and the budesonide+

Xiao

’

er Jianpi Yifei

Formula group . Compared to the budesonide group

the expression of HMGB1 in the

Xiao

’

er Jianpi Yifei

Formula group was decreased significantly

while the expression of both TLR4 and MyD88 was increased

and the expressions of HMGB1

TLR4

and MyD88 in the budesonide+

Xiao

’

er Jianpi Yifei

Formula group were decreased significantly

with statistically significant differences(

0.05). The IF result showed that HMGB1 expression levels in the

Xiao

’

er Jianpi Yifei

Formula group

budesonide group

and budesonide+

Xiao

’

er Jianpi Yifei

Formula group were significantly lower than that in the model group(

0.01)

and the HMGB1 and TLR4 expression levels in the budesonide+

Xiao

’

er Jianpi Yifei

Formula group were significantly lower than those in the budesonide group(

0.05).

Conclusion

Xiao

’

er Jianpi Yifei

Formula and budesonide could effectively alleviate asthma-related pathological changes

reduce the expression of inflammatory cytokines

inhibit the HMGB1/TLR4/NF-κB inflammatory signaling pathway

and improve airway hyperresponsiveness. The combination of

Xiao

’

er Jianpi Yifei

Formula and budesonide was superior to budesonide alone. The HMGB1/TLR4/NF-κB inflammatory signaling pathway may be a potential mechanism for

Xiao

’

er Jianpi

Yifei Formula to improve airway hyperresponsiveness in children with asthma.

关键词

Keywords

references

中华儿科杂志编辑委员会，中华医学会儿科学分会呼吸学组，中国医师协会儿科医师分会儿童呼吸专业委员会．儿童支气管哮喘规范化诊治建议(2020年版) [J]. 中华儿科杂志， 2020 , 58 ( 9 ): 708 － 717 .

Global Initiative for Asthma . Global Strategy for Asthma Management and Prevention [EB/OL].( 2023－05－02 )[ 2023－06－26 ]. https：//ginasthma.org/wp-content/uploads/2023/05/GINA－2023－Full-Report－2023－WMS.pdf .

肖惠迪，书文，李梦龙，等 . 中国2011—2018年儿童哮喘患病率Meta分析 [J]. 中国学校卫生， 2020 , 41 ( 8 ): 1208 － 1211 .

ELISEEVA TI , GEPPE NA , TUSH EV , et al . Body Height of Children with Bronchial Asthma of Various Severities [J/OL]. Can Respir J , 2017 , 2017 ： 8761404 [ 2022－11－08 ]. https：//www.hindawi.com/journals/crj/2017/8761404/ https：//www.hindawi.com/journals/crj/2017/8761404/ .

ULLAH MA , LOH Z , GAN WJ , et al . Receptor for advanced glycation end products and its ligand high-mobility group box－1 mediate allergic airway sensitization and airway inflammation [J]. J Allergy Clin Immunol , 2014 , 134 ( 2 ): 440 － 450 .

ZHOU Y , JIANG YQ , WANG WX , et al . HMGB1 and RAGE levels in induced sputum correlate with asthma severity and neutrophil percentage [J]. Hum Immunol , 2012 , 73 ( 11 ): 1171 － 1174 .

SHALABY KH , HEIALY SA , TSUCHIYA K , et al . The TLR4－TRIF pathway can protect against the development of experimental allergic asthma [J]. Immunology , 2017 , 152 ( 1 ): 138 － 149 .

ZHANG H , YANG N , WANG TY , et al . Vitamin D reduces inflammatory response in asthmatic mice through HMGB1/TLR4/NF－κB signaling pathway [J]. Mol Med Rep , 2018 , 17 ( 2 ): 2915 － 2920 .

刘礼荣，符诒慧，周黎明，等 . 抑制HMGB1对支气管哮喘小鼠气道炎症的影响及机制 [J]. 中华医院感染学杂志， 2022 , 32 ( 11 ): 1632 － 1637 .

姚如婕，刘沉涛，黄榕，等 . 布地奈德雾化治疗对哮喘小鼠糖皮质激素受体及核因子－κB表达的影响 [J]. 中国当代儿科杂志， 2015 , 17 ( 1 ): 86 － 89 .

KANG JY , LEE SY , RHEE CK , et al . Effect of aging on airway remodeling and muscarinic receptors in a murine acute asthma model [J]. Clin Interv Aging , 2013 , 8 ： 1393 － 1403 .

PADRID P , SNOOK S , FINUCANE T , et al . Persistent airway hyperresponsiveness and histologic alterations after chronic antigen challenge in cats [J]. Am J Respir Crit Care Med , 1995 , 151 ( 1 ): 184 － 193 .

高桂玲 . 参苓白术散辅助丙酸氟替卡松气雾剂治疗肺脾气虚型哮喘缓解期患儿的效果 [J]. 临床医学， 2022 , 42 ( 8 ): 115 － 157 .

邬素珍，莫珊，陆柳丹，等 . 健脾补肺膏对咳嗽变异性哮喘缓解期儿童的影响 [J]. 中国中西医结合杂志， 2023 ， 43 ( 1 )： 113 － 116 .

杨宏志，郭光辉，杨俊姝，等 . 益肺Ⅱ号方对支气管哮喘缓解期免疫功能和血清炎症因子的影响 [J]. 中华中医药学刊， 2020 , 38 ( 4 ): 94 － 97 .

吴振奎，钟丽花，刘延霞，等 . 女贞子多糖联合布地奈德在哮喘幼鼠模型中的免疫调节及抗氧化作用研究 [J]. 中国免疫学杂志， 2022 , 38 ( 9 ): 1069 － 1074 .

HOU CC , KONG JC , LIANG Y , et al . HMGB1 contributes to allergen-induced airway remodeling in a murine model of chronic asthma by modulating airway inflammation and activating lung fibroblasts [J]. Cell Mol Immunol , 2015 , 12 ( 4 ): 409 － 423 .

LV YH , LI YL , ZHANG DD , et al . HMGB1－induced asthmatic airway inflammation through GRP75－mediated enhancement of ER-mitochondrial Ca 2＋ transfer and ROS increased [J]. J Cell Biochem , 2018 , 119 ( 5 ): 4205 － 4215 .

DI CANDIA L , GOMEZ E , VENEREAU E , et al . HMGB1 is upregulated in the airways in asthma and potentiates airway smooth muscle contraction via TLR4 [J]. J Allergy Clin Immunol , 2017 , 140 ( 2 ): 584 － 587 .

HOU CC , ZHAO HJ , LIU LY , et al . High mobility group protein B1(HMGB1) in Asthma: comparison of patients with chronic obstructive pulmonary disease and healthy controls [J]. Mol Med , 2011 , 17 ( 7/8 ): 807 － 815 .

OTTOW MK , KLAVER EJ , VAN DER POUW KRAAN TC , et al . The helminth Trichuris suis suppresses TLR4－induced inflammatory responses in human macrophages [J]. Genes Immun , 2014 , 15 ( 7 ): 477 － 486 .

KIM HS , CHUNG DH . TLR4－mediated IL－12 production enhances IFN－γ and IL－1β production, which inhibits TGF－β production and promotes antibody-induced joint inflammation [J/OL]. Arthritis Res Ther , 2012 , 14 ( 5 ): R210 [ 2022－11－08 ]. https：//arthritis-research.biomedcentral.com/articles/10.1186/ar4048 https：//arthritis-research.biomedcentral.com/articles/10.1186/ar4048 .

ZHU HT , BIAN C , YUAN JC , et al . Curcumin attenuates acute inflammatory injury by inhibiting the TLR4/MyD88/NF－κB signaling pathway in experimental traumatic brain injury [J/OL]. J Neuroinflammation , 2014 , 11 ： 59 [ 2022－11－08 ]. https：//jneuroinflammation.biomedcentral.com/articles/10.1186/1742－2094－11－59 https：//jneuroinflammation.biomedcentral.com/articles/10.1186/1742－2094－11－59 .

乔俊英，宋丽，张艳丽，等 . 哮喘小鼠HMGB1/TLR4/NF－κB信号通路及维生素D的作用 [J]. 中国当代儿科杂志， 2017 , 19 ( 1 ): 95 － 103 .

LEE CC , LAI YT , CHANG HT , et al . Inhibition of high-mobility group box 1 in lung reduced airway inflammation and remodeling in a mouse model of chronic asthma [J]. Biochem Pharmacol , 2013 , 86 ( 7 ): 940 － 949 .

SHANG LQ , WANG L , SHI XL , et al . HMGB1 was negatively regulated by HSF1 and mediated the TLR4/MyD88/NF－κB signal pathway in asthma [J/OL]. Life Sci , 2020 , 241 ： 117120 [ 2022－11－08 ]. http：// www.sciencedirect.com/science/article/abs/pii/S0024320519310-471？via%3Dihub http：// www.sciencedirect.com/science/article/abs/pii/S0024320519310-471？via%3Dihub .

何苗 . 布地奈德对高氧致新生小鼠支气管肺发育不良及HMGB1/TLR4/NF－κB通路的影响 [J]. 中国医师杂志， 2019 , 21 ( 8 ): 1199 － 1204 .

刘虹，王洁英，刘娜，等 . 布地奈德混悬液对哮喘模型小鼠肺组织TLR4/MyD88/NF－κB通路的影响 [J]. 现代生物医学进展， 2021 , 21 ( 23 ): 4432 － 4435 .

李星，李竹英，王珏 . 平喘颗粒对哮喘气道相关炎症因子及肺组织HMGB1、TLR4表达的影响 [J]. 中国中医急症， 2021 , 30 ( 6 ): 941 － 944 .

程哲，代灵灵，曹德飞，等．支气管哮喘患者诱导痰中HMGB1和RAGE水平的变化及临床意义 [J] . 中华医学杂志， 2011 ， 91 ( 22 ): 1538 － 1542 .

DONG YM , YAN H , ZHAO X , et al . Gu-Ben-Fang-Xiao Decoction Ameliorated Murine Asthma in Remission Stage by Modulating Microbiota-Acetate-Tregs Axis [J/OL]. Front Pharmacol , 2020 , 11 ： 549 [ 2022－11－08 ]. https：//www.frontiersin.org/articles/10.3389/fphar.2020.00549/full https：//www.frontiersin.org/articles/10.3389/fphar.2020.00549/full .

陈国榕，吴东 . 肠道菌群代谢产物丁酸减轻重症急性胰腺炎相关肺损伤的机制 [J]. 中华临床营养杂志， 2020 ， 28 ( 6 )： 363 － 367 .

吴玉苗，吴要伟，朱万青，等 . 短链脂肪酸与儿童哮喘的关系及治疗研究进展 [J]. 中国现代医学杂志， 2022 ， 32 ( 16 )： 37 － 42 .

丁泽君，孙明洁，刘佳，等．丁酸钠对脂多糖诱导的急性肺损伤大鼠的肺保护研究 [J]. 国际麻醉学与复苏杂志， 2013 ， 34 ( 9 )： 779 － 782 .

LI N , LIU XX , HONG M , et al . Sodium butyrate alleviates LPS-induced acute lung injury in mice via inhibiting HMGB1 release [J]. Int Immunopharmacol , 2018 , 56 ： 242 － 248 .

Views

1221

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect and mechanism of Xiao' er Zhixiao Pingchuan Granules in improving airway remodeling in asthmatic mice

Pregnancy preservation mechanism of Shoutai Pills in promoting decidualization by delaying endometrial stromal cell senescence

Study of the mechanism of electroacupuncture for alleviating chronic hyperalgesia in mice through modulation of spinal dorsal horn microglia

Bushen Tiaojing Formula on ovarian granulosa cell proliferation and steroid hormone secretion in aged mice by regulating LIF and its receptor

Study on electroacupuncture at "Ciliao" (BL32) improving stress urinary incontinence in mice by promoting endogenous bone marrow mesenchymal stem cell homing

Related Author

CHEN Xun

AN Zimeng

LI Min

WAN Tong

HUANG Jing

JI Xiaohua

YAN Xiaoru

ZHANG Luyi

Related Institution

中国中医科学院西苑医院北京 100091Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091, China

Hebei University of Chinese Medicine

Hebei Key Laboratory of Integrated Traditional Chinese and Western Medicine for Liver and Kidney Diseases

Hebei Collaborative Innovation Center for Integrated Traditional Chinese and Western Medicine in Reproductive Diseases

Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences

AI问答

Postal code：100029
Tel：(010)53911857 Email：jbutcm@bucm.edu.cn
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰

Experimental study of Xiao’er Jianpi Yifei Formula improving airway hyperresponsiveness in asthmatic mice

DOI：10.3969/j.issn.1006-2157.2023.09.012

摘要

Abstract

关键词

Keywords

references