清肺化痰汤通过抑制JAK2/STAT3通路上调CFTR的表达治疗COPD

屈飞; 邱沙英; 张明月; 黄俊浩; 曾杰; 刘婕; 崔艳茹

doi:10.13422/j.cnki.syfjx.20201602

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清肺化痰汤通过抑制JAK2/STAT3通路上调CFTR的表达治疗COPD

经典名方 | 更新时间：2020-09-09

- 清肺化痰汤通过抑制JAK2/STAT3通路上调CFTR的表达治疗COPD
- Efficacy of Qingfei Huatantang in Treatment of COPD by Up-regulating Expression of CFTR Through JAK2/STAT3 Pathway
- 中国实验方剂学杂志 2020年26卷第18期页码：1-7
- 作者机构：
  
  1.江西中医药大学药学院，江西省中药药理学重点实验室，南昌 330004
  2.江西中医药大学中医学院，南昌 330004
- 作者简介：
  
  屈飞，博士，副教授，硕士生导师，从事中药药理学研究，E-mail：quf0917@163.com
  崔艳茹，硕士，副教授，硕士生导师，从事中药药理学研究，Tel：0791-87118930，E-mail：cuiyanru2@163.com
- 基金信息：
  
  国家自然科学基金项目(81760744);江西省教育厅科技重点项目(GJJ190633)
- DOI：10.13422/j.cnki.syfjx.20201602
  中图分类号： R2-0;R22;R285.5;R289
- 收稿日期：2019-12-11，
  
  网络出版日期：2020-06-05，
  
  纸质出版日期：2020-09-20
- 稿件说明：
移动端阅览
屈飞,邱沙英,张明月等.清肺化痰汤通过抑制JAK2/STAT3通路上调CFTR的表达治疗COPD[J].中国实验方剂学杂志,2020,26(18):1-7.

QU Fei,QIU Sha-ying,ZHANG Ming-yue,et al.Efficacy of Qingfei Huatantang in Treatment of COPD by Up-regulating Expression of CFTR Through JAK2/STAT3 Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(18):1-7.
屈飞,邱沙英,张明月等.清肺化痰汤通过抑制JAK2/STAT3通路上调CFTR的表达治疗COPD[J].中国实验方剂学杂志,2020,26(18):1-7. DOI： 10.13422/j.cnki.syfjx.20201602.

QU Fei,QIU Sha-ying,ZHANG Ming-yue,et al.Efficacy of Qingfei Huatantang in Treatment of COPD by Up-regulating Expression of CFTR Through JAK2/STAT3 Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(18):1-7. DOI： 10.13422/j.cnki.syfjx.20201602.

摘要

目的

探索清肺化痰汤对慢性阻塞性肺疾病（COPD）的治疗作用及其机制研究。

方法

应用烟雾吸入联合脂多糖（LPS）灌肺复合素的方法，建立COPD大鼠模型，造模成功后将大鼠随机分为6组，分别为正常组，COPD模型组，清肺化痰汤低、中、高剂量组和氨溴索组。造模28 d后开始给药，清肺化痰汤低、中、高剂量7.5，15，30 g·kg

-1

，氨溴索35 mg·kg

-1

，连续给药14 d。免疫组化和实时荧光定量聚合酶链式反应（Real-time PCR）检测肺组织囊性纤维化跨膜转导调节因子（CFTR）蛋白和mRNA的表达。应用LPS诱导NCI-H292细胞建立黏液高分泌模型，实验分为8组，分别为空白组，LPS组，LPS+10%胎牛血清，LPS+生理血清，LPS+5%含药血清，LPS+10%含药血清，LPS+20%含药血清，LPS+AG490组。免疫荧光、蛋白免疫印迹法（Western blot）和Real-time PCR观察LPS刺激后的NCI-H292细胞中CFTR蛋白和mRNA的表达，Western blot检测LPS刺激后的NCI-H292细胞中Janus激酶2/信号转导与转录激活因子3（JAK2/STAT3）信号通路表达。

结果

正常组大鼠肺组织管腔周围有大量棕褐色颗粒，COPD表达升高，与正常组比较，模型组大鼠肺组织管腔周围的棕褐色颗粒极少，COPD表达较低。与正常组比较，COPD模型组大鼠肺组织中CFTR mRNA和蛋白表达明显降低（

0.05）；与模型组比较，清肺化痰汤低、中、高剂量组明显升高大鼠肺组织CFTR mRNA和蛋白表达水平（

0.05）。与空白组比较，LPS组NCI-H292细胞CFTR mRNA和蛋白表达显著降低（

0.01），p-JAK2，p-STAT3蛋白表达明显升高（

0.05）；与模型组比较，清肺化痰汤5%，10%，20%含药血清组明显升高CFTR mRNA和蛋白表达，明显降低p-JAK2，p-STAT3蛋白表达（

0.05，

0.01）。

结论

清肺化痰汤通过抑制JAK2/STAT3通路来上调CFTR治疗COPD。

Abstract

Objective

To explore the efficacy and mechanism of Qingfei Huatan Tang on chronic obstructive pulmonary disease (COPD).

Method

The rat model of COPD was established through smoke inhalation combined with lipopolysaccharide (LPS) and pulmonary compound injection. After successful modeling

the rats were randomly divided into 6 groups

namely the control group

the COPD model group

low

medium and high-dose Qingfei Huatan Tang groups and the ambroxol group. After 28 days of modeling

the drug was administered. Low

medium and high-dose Qingfei Huatan Tang (7.5

30 g·kg

-1

) and ambroxol (35 mg·kg

-1

) were administered continuously for 14 days. Immunohistochemistry and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect protein expression and mRNA expression of cystic fibrosis transmembrane conductance regulator (CFTR) in pulmonary fibrosis. NCI-H292 cells were induced by LPS to establish a mucus hypersecretion model. The experiment was divided into 8 groups

namely the blank control group

LPS group

LPS+10% fetal bovine serum group

LPS+ physiological serum group

LPS+5% drug serum group

LPS+10% drug serum group

LPS+20% drug serum group and LPS+AG490 group. Immunofluorescence

Western blot and Real-time PCR were used to observe the protein and mRNA expressions of CFTR in NCI-H292 cells after LPS stimulation

and western blot was used to detect the expression of tyrosine kinase 2/transcription factor 3 (JAK2/STAT3) signaling pathway in NCI-H292 cells after LPS stimulation.

Result

There were a large number of brown particles around the lumen of lung tissues in the normal group

with increased COPD expression. There were a few brown particles around the lumen of lung tissues in the model group compared with the normal group

with decreased COPD expression. Compared with the normal group

mRNA and protein expressions of CFTR in the lung tissues of the COPD model group were significantly decreased (

0.05). Compared with the model group

mRNA and protein expressions of CFTR in the lung tissues of low

medium and high-dose Qingfei Huatan Tang groups (

0.05). Compared with the blank control group

mRNA and protein expressions of CFTR in NCI-H292 cells of the LPS group (

0.05)

with significant increases in protein expressions of p-JAK2 and p-STAT3 (

0.05). Compared with the model group

10%

20% Qingfei Huatan Tang-containing serum groups showed significant increases in mRNA and protein expressions of CFTR

but with significant decreases in p-JAK2

p-STAT3 protein expressions (

0.05

0.01).

Conclusion

Qingfei Huatan Tang up-regulated CFTR in the treatment of COPD by inhibiting JAK2/STAT3 pathway.

关键词

Keywords

references

VESTBO J , HURD S S , AGUSTÍ A G , et al . Global Strategy for the diagnosis,management,and prevention of chronic obstructive pulmonary disease: GOLD executive summary [J]. Am J Respir Crit Care Med , 2013 , 187 ( 4 ): 347 - 365 .

KHAKBAN A , SIN D D , FITZGERALD J M , et al . The projected epidemic of chronic obstructive pulmonary disease hospitalizations over the next 15 years.A population-based perspective [J]. Am J Respir Crit Care Med , 2017 , 195 ( 3 ): 287 - 291 .

BARNES P J . Chronic obstructive pulmonary disease: a growing but neglected global epidemic [J]. PLoS Med , 2007 , 4 ( 5 ): e112 .

XIE C ， TANG X ， XU W ， et al . A host defense mechanism involving CFTR-mediated bicarbonate secretion in bacterial prostatitis [J]. PLoS One , 2010 , 5 ( 12 ): 1 - 7 .

李建生，王至婉，余学庆 , 等 . 中药治疗慢性阻塞性肺疾病稳定期的系统评价 [J]. 辽宁中医杂志 , 2010 , 37 ( 2 ): 229 - 231 .

彭波，李泽庚，童佳兵 , 等 . 慢性阻塞性肺疾病中医药研究进展 [J]. 辽宁中医药大学学报 , 2011 , 13 ( 6 ): 36 - 38 .

陈斯宁 , 徐贞 , 黄美杏 . 清肺化痰汤对COPD急性加重期痰热郁肺证患者细胞因子的影响 [J]. 云南中医中药杂志 , 2008 , 29 ( 9 ): 6 .

屈飞 , 黄小婷 , 崔艳茹 , 等 . 清肺化痰汤对慢性阻塞性肺疾病大鼠模型气道炎症和黏液分泌的调控作用研究 [J]. 中华中医药学刊 , 2019 , 37 ( 9 ): 2064 - 2067 .

宋小莲 , 王昌惠 , 白冲 . 脂多糖结合熏烟法和单纯熏烟法建立慢性阻塞性肺病大鼠模型的比 [J]. 第二军医大学学报 , 2010 , 31 ( 3 ): 246 - 249 .

YAN X , SONG Y , SHEN C , et al . Mucoactive and antioxidant medicines for COPD:consensus of a group of Chinese pulmonary physicians [J]. Int J Chron Obstruct Pulmon Dis , 2017 , 12 : 803 - 812 .

朱仕兵 , 许安 , 张德文 . 清肺化痰汤对老年慢性阻塞性肺疾病急性加重期合并呼吸衰竭患者肺功能、炎症因子及血气分析的影响 [J]. 中国老年学杂志 , 2017 , 37 ( 19 ): 4838 - 4840 .

余彩云 , 杜永明 , 陈国宁 , 等 . 清肺化痰汤雾化对慢性阻塞性肺疾病急性加重期的临床运用分析 [J]. 中国中医基础医学杂志 , 2014 , 20 ( 5 ): 694 - 696 .

屈飞 , 崔艳茹 , 李惠兰 . 清肺化痰汤抗COPD大鼠模型黏蛋白作用研究 [J]. 亚太传统医药 , 2016 , 12 ( 10 ): 11 - 13 .

KESIMER M , FORD A A , CEPPE A , et al . Airway mucin concentration as a marker of chronic bronchitis [J]. N Engl J Med , 2017 , 377 ( 10 ): 911 - 922 .

MARTIN C , FRIJA-MASSON J , BURGEL P R . Targetingmucus hypersecretion: new therapeutic opportunities for COPD? [J]. Drugs , 2014 , 74 ( 10 ): 1073 - 1089 .

STOLTZ D A , MEYERHOLZ D K , WELSH M J . Origins of cystic fibrosis lung disease [J]. N Engl J Med , 2015 , 372 : 1574 - 1575 .

PEZZULO A A , TANG X X , HOEGGER M J , et al . Reduced airway surface pH impairs bacterial killing in the porcine cystic fibrosis lung [J]. Nature , 2012 , 487 : 109 - 113 .

ROY M G , LIVRAGHI-BUTRICO A , FLETCHER A A , et al . Muc5b is required for airway defence [J]. Nature , 2014 , 505 : 412 - 416 .

HOEGGER M J , FISCHER A J , MCMENIMEN J D , et al . Impaired mucus detachment disrupts mucociliary transport in a piglet model of cystic fibrosis [J]. Science , 2014 , 345 : 818 - 822 .

BARNES P J , BURNEY P G , SILVERMAN E K , et al . Chronic obstructive pulmonary disease [J]. Nat Rev Dis Primers , 2015 , 1 : 15076 .

RAJU S V , LIN V Y , LIU L , et al . The cystic fibrosis transmembrane conductance regulator potentiator ivacaftor augments mucociliary clearance abrogating cystic fibrosis transmembrane conductance regulator inhibition by cigarette smoke [J]. Am J Respir Cell Mol Biol , 2017 , 56 ( 1 ): 99 - 108 .

QU F , QIN X Q , CUI Y R , et al . Ozone stress down-regulates the expression of cystic fibrosis transmembrane conductance regulator in human bronchial epithelial cells [J]. Chem Biol Interact , 2009 , 179 : 219 - 226 .

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