Mechanism of Number 2 Feibi Recipe in Ameliorating Pulmonary Fibrosis in Mice by Modulating Endoplasmic Reticulum Stress in AT2 Cells to Attenuate Apoptosis and Promote Alveolar Repair

CAI Yaodong; BEI Jialing; WEI Wan; XU Chengyan; LIU Yanli; WANG Yong; JIAO Yang; CHEN Yun

doi:10.13422/j.cnki.syfjx.20252509

您当前的位置：

首页 >

文章列表页 >

Mechanism of Number 2 Feibi Recipe in Ameliorating Pulmonary Fibrosis in Mice by Modulating Endoplasmic Reticulum Stress in AT2 Cells to Attenuate Apoptosis and Promote Alveolar Repair

更新时间：2026-04-28

- Mechanism of Number 2 Feibi Recipe in Ameliorating Pulmonary Fibrosis in Mice by Modulating Endoplasmic Reticulum Stress in AT2 Cells to Attenuate Apoptosis and Promote Alveolar Repair
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 32, Issue 10, Pages: 80-92(2026)
- 作者机构：
  
  1.北京中医药大学第二临床医学院，北京 100078
  2.北京中医药大学中医学院，北京 100029
  3.北京中医药大学东方医院，北京 100078
  4.北京中医药大学东直门医院，北京 100700
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20252509
  CLC： R289;R259;R285
- Received：25 September 2025，
  
  Revised：2026-01-07，
  
  Accepted：07 January 2026，
  
  Online First：08 January 2026，
  
  Published：20 May 2026
- 稿件说明：
移动端阅览
蔡耀东,贝佳凌,韦婉等.肺痹汤2号调控AT2细胞内质网应激抵抗凋亡促进肺泡修复改善小鼠肺纤维化的机制[J].中国实验方剂学杂志,2026,32(10):80-92.

CAI Yaodong,BEI Jialing,WEI Wan,et al.Mechanism of Number 2 Feibi Recipe in Ameliorating Pulmonary Fibrosis in Mice by Modulating Endoplasmic Reticulum Stress in AT2 Cells to Attenuate Apoptosis and Promote Alveolar Repair[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(10):80-92.
蔡耀东,贝佳凌,韦婉等.肺痹汤2号调控AT2细胞内质网应激抵抗凋亡促进肺泡修复改善小鼠肺纤维化的机制[J].中国实验方剂学杂志,2026,32(10):80-92. DOI： 10.13422/j.cnki.syfjx.20252509.

CAI Yaodong,BEI Jialing,WEI Wan,et al.Mechanism of Number 2 Feibi Recipe in Ameliorating Pulmonary Fibrosis in Mice by Modulating Endoplasmic Reticulum Stress in AT2 Cells to Attenuate Apoptosis and Promote Alveolar Repair[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(10):80-92. DOI： 10.13422/j.cnki.syfjx.20252509.

摘要

目的

探讨中药复方肺痹汤2号在特发性肺纤维化中的干预机制，聚焦其对肺泡Ⅱ型上皮细胞（AT2）内质网应激、凋亡、干性维持及再生修复能力的影响，验证“宗气衰微-肺泡痿痹”理论的现代转化路径。

方法

采用博来霉素（BLM）诱导小鼠肺纤维化模型，设空白组，模型组与肺痹汤2号低、高剂量组（9.1、18.2 g·kg

-1

）及醋酸波尼松龙组（6.5 mg·kg

-1

）。通过苏木素-伊红（HE）及马松（Masson）染色评估肺组织结构变化及胶原沉积情况，采用碱水解法测量羟脯氨酸（HYP）含量，测量肺系数及肺功能指标。应用实时荧光定量聚合酶链式反应（Real-time PCR）检测纤维化相关因子［胶原蛋白Ⅰ型

1链（ColⅠa1）、

平滑肌肌动蛋白（

α-

SMA）、金属蛋白酶组织抑制剂1（Timp1）］mRNA表达水平。使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL）评估细胞凋亡，表面活性蛋白C（SPC）和半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）双染免疫荧光法进一步评估AT2细胞凋亡，SPC和蛋白激酶R样内质网激酶（PERK）双染检测AT2细胞内质网应激情况，透射电镜观察AT2细胞内质网及板层小体超微结构变化。通过蛋白免疫印迹法（Western blot）检测内质网应激及凋亡通路关键蛋白PERK、激活转录因子4（Atf4）及Ca

spase-3表达情况。通过双染免疫荧光标记SPC和Ki-67抗原（Ki-67）分析AT2细胞增殖能力，利用谱系追踪技术（GFP

小鼠）联合Krt8标记检测分化中间状态，并观察AT2细胞向肺泡Ⅰ型上皮细胞（AT1）细胞形态的转化。

结果

BLM诱导后小鼠出现显著肺组织结构破坏、胶原沉积增加、肺系数升高、肺功能下降和纤维化因子上调（

0.01）。高剂量肺痹汤2号干预可显著改善肺组织损伤与功能障碍，显著降低HYP含量（

0.01），显著下调ColⅠa1、

-SMA、Timp1表达（

0.01）。凋亡分析显示模型组TUNEL阳性细胞比例上升，AT2细胞Caspase-3阳性率显著增加，而高剂量组显著降低。透射电镜下AT2细胞内质网结构肿胀，给药治疗后趋于正常；PERK蛋白质染色分析发现模型组AT2细胞内质网应激明显，给药组显著缓解。模型组中内质网应激相关蛋白PERK、Atf4及凋亡蛋白Caspase-3的表达升高，给药后可显著降低。透射电镜下模型组板层小体结构紊乱，治疗组趋于恢复。AT2细胞增殖能力方面，治疗组Ki-67⁺SPC⁺细胞比例显著上升（

0.01）。谱系追踪显示模型组角蛋白8阳性绿色荧光蛋白阳性（Krt8⁺GFP⁺）细胞占比升高，提示分化阻滞；治疗组该比例显著下降，同时GFP⁺细胞形态向扁平延展转变，提示分化向AT1方向恢复。

结论

肺痹汤2号通过缓解AT2细胞内质网应激、减少AT2细胞凋亡、恢复其板层小体结构与功能、增强其增殖活性、缓解分化阻滞促进其向AT1细胞分化，从而修复肺泡有效阻断肺纤维化的进展。其“复宗气、和气血、通肺络”的中医治疗机制与现代AT2干细胞调控路径高度契合，为中药干预IPF提供了新理论与实验依据。

Abstract

Objective

To investigate the intervention mechanism of the traditional Chinese medicine Number 2 Feibi recipe （N2FBR） in idiopathic pulmonary fibrosis （IPF）， focusing on its effects on endoplasmic reticulum （ER） stress， apoptosis， stemness maintenance， and regenerative capacity of alveolar type Ⅱ epithelial cells （AT2 cells）， and to validate the modern translational pathway of the theory of "deficiency of Zong Qi leading to pulmonary atelectasis and atrophy".

Methods

A mouse model of pulmonary fibrosis was induced by bleomycin （BLM）. Mice were randomly divided into blank control， model， low-， and high-dose N2FBR intervention groups （9.1， 18.2 g·kg

-1

）， and prednisolone intervention group （6.5 mg·kg

-1

）. Pulmonary histopathological changes and collagen deposition were evaluated using hematoxylin-eosin （HE） and Masson's trichrome staining. Hydroxyproline （HYP） content was measured by the alkaline hydrolysis method. Lung coefficient and pulmonary function parameters were evaluated. The

mRNA expression levels of fibrosis-related factors， including collagen type Ⅰ alpha 1 chain （ColIa1）， alpha-smooth muscle actin （

-SMA）， and tissue inhibitor of metalloproteinase 1 （Timp1）， were detected by real-time polymerase chain reaction （Real-time PCR）. Cell apoptosis was assessed using the terminal deoxynucleotidyl transferase dUTP nick-end labeling （TUNEL） assay. Apoptosis of AT2 cells was further evaluated by double immunofluorescence staining for surfactant protein C （SPC） and cysteine-aspartic protease-3 （Caspase-3）. Endoplasmic reticulum （ER） stress in AT2 cells was examined by double staining for SPC and protein kinase R-like endoplasmic reticulum kinase （PERK）. Ultrastructural changes of ER and lamellar bodies in AT2 cells were observed by transmission electron microscopy （TEM）. The expression levels of key proteins involved in ER stress and apoptosis pathways， including PERK， activating transcription factor 4 （ATF4）， and Caspase-3， were detected by Western blot. Double immunofluorescence staining of SPC and Ki-67 antigen （Ki-67） was performed to evaluate the proliferative capacity of AT2 cells. Lineage tracing technology （labeling AT2 cells with GFP） combined with Krt8 labeling was used to evaluate intermediate differentiation states， and morphological transformation of AT2 cells into alveolar type Ⅰ epithelial cells （AT1） was observed.

Results

BLM-induced mice exhibited significant structural disruption of lung tissue， increased collagen deposition， elevated lung coefficient， decreased pulmonary function， and upregulation of fibrosis-related factors （

0.01）. High-dose N2FBR treatment significantly ameliorated lung tissue damage and dysfunction， significantly reduced HYP content （

0.01）， and significantly downregulated ColIa1，

-SMA， and Timp1 expression （

0.01）. Apoptosis analysis showed increased TUNEL-positive and Caspase-3-positive AT2 cells in the model group， which w

as significantly reduced by high-dose N2FBR treatment. TEM revealed swollen ER structures in AT2 cells of the model group， which tended to return to normal following treatment. PERK protein staining analysis showed evident ER stress in AT2 cells of the model group， which were markedly alleviated in the treatment group. The expression levels of ER stress-related proteins PERK and ATF4， as well as the apoptosis-related protein Caspase-3， were elevated in the model group and significantly reduced after treatment. TEM also revealed disrupted lamellar body structures in the model group， which tended to recover in the treatment group. Regarding the proliferative capacity of AT2 cells， the proportion of Ki-67⁺SPC⁺ AT2 cells significantly increased in the treatment group （

0.01）. Lineage tracing showed that the proportion of keratin 8-positive green fluorescent protein-positive （Krt8⁺GFP⁺） cells increased in the model group， indicating differentiation arrest. This proportion was significantly reduced in the treatment group， and the morphology of GFP⁺ cells exhibited a flattened， extended shape， suggesting restored differentiation toward AT1 cells.

Conclusion

N2FBR alleviates ER stress in AT2 cells， reduces AT2 cell apoptosis， restores lamellar body structure and function， enhances proliferation activity， and alleviates differentiation arrest to promote differentiation into AT1 cells， thereby repairing the alveolar epithelium and effectively blocking the progression of pulmonary fibrosis. Its traditional Chinese medicine mechanism of "replenishing Zong Qi， harmonizing Qi and blood， and unblocking pulmonary meridians" closely aligns with the modern regulatory pathway of AT2 stem cells， providing a novel theoretical basis and experimental evidence for the intervention of IPF with traditional Chinese medicine.

关键词

Keywords

references

GLASS D S ， GROSSFELD D ， RENNA H A ， et al . Idiopathic pulmonary fibrosis：Current and future treatment ［J］. Clin Respir J ， 2022 ， 16 （ 2 ）： 84 - 96 .

DEMPSEY T M ， SANGARALINGHAM L R ， YAO X ， et al . Clinical effectiveness of antifibrotic medications for idiopathic pulmonary fibrosis ［J］. Am J Respir Crit Care Med ， 2019 ， 200 （ 2 ）： 168 - 174 .

GUILLOT L ， NATHAN N ， TABARY O ， et al . Alveolar epithelial cells：Master regulators of lung homeostasis ［J］. Int J Biochem Cell Biol ， 2013 ， 45 （ 11 ）： 2568 - 2573 .

WANG Y ， WANG L ， MA S ， et al . Repair and regeneration of the alveolar epithelium in lung injury ［J］. FASEB J ， 2024 ， 38 （ 8 ）： e23612 .

PARIMON T ， YAO C ， STRIPP B R ， et al . Alveolar epithelial type II cells as drivers of lung fibrosis in idiopathic pulmonary fibrosis ［J］. Int J Mol Sci ， 2020 ， 21 （ 7 ）： 2269 .

BARKAUSKAS C E ， CRONCE M J ， RACKLEY C R ， et al . Type 2 alveolar cells are stem cells in adult lung ［J］. J Clin Invest ， 2013 ， 123 （ 7 ）： 3025 - 3036 .

LIU K ， MENG X ， LIU Z ， et al . Tracing the origin of alveolar stem cells in lung repair and regeneration ［J］. Cell ， 2024 ， 187 （ 10 ）： 2428 - 2445 e20 .

LIU S ， XI Q ， LI X ， et al . Mitochondrial dysfunction and alveolar type Ⅱ epithelial cell senescence：The destroyer and rescuer of idiopathic pulmonary fibrosis ［J］. Front Cell Dev Biol ， 2025 ， 13 ： 1535601 .

WANG Z ， WEI D D ， BIN E N ， et al . Enhanced glycolysis-mediated energy production in alveolar stem cells is required for alveolar regeneration ［J］. Cell Stem Cell ， 2023 ， 30 （ 8 ）： 1028 - 1042.e7 .

WU H J ， YU Y Y ， HUANG H W ， et al . Progressive pulmonary fibrosis is caused by elevated mechanical tension on alveolar stem cells （vol 180，pg 107，2020）［J］. Cell ， 2021 ， 184 （ 3 ）： 845 - 846 .

SUN Z ， HE W ， MENG H ， et al . Lactate activates ER stress to promote alveolar epithelial cells apoptosis in pulmonary fibrosis ［J］. Respir Res ， 2024 ， 25 （ 1 ）： 401 .

KLYMENKO O ， HUEHN M ， WILHELM J ， et al . Regulation and role of the ER stress transcription factor CHOP in alveolar epithelial type-Ⅱ cells ［J］. J Mol Med （Berl）， 2019 ， 97 （ 7 ）： 973 - 990 .

SHIN S Y ， CHEN J ， MILMAN KRENTSIS I ， et al . From epithelium to therapy：Transitional cells in lung fibrosis ［J］. Am J Respir Cell Mol Biol ， 2025 ， 72 （ 5 ）： 472 - 483 .

KOBAYASHI Y ， TATA A ， KONKIMALLA A ， et al . Persistence of a regeneration-associated，transitional alveolar epithelial cell state in pulmonary fibrosis ［J］. Nat Cell Biol ， 2020 ， 22 （ 8 ）： 934 - 946 .

ZHAO R ， WANG Z ， WANG G ， et al . Sustained amphiregulin expression in intermediate alveolar stem cells drives progressive fibrosis ［J］. Cell Stem Cell ， 2024 ， 31 （ 9 ）： 1344 - 1358 .

ENOMOTO Y ， KATSURA H ， FUJIMURA T ， et al . Autocrine TGF-beta-positive feedback in profibrotic AT2-lineage cells plays a crucial role in non-inflammatory lung fibrogenesis ［J］. Nat Commun ， 2023 ， 14 （ 1 ）： 4956 .

付小芳，吴志松，曹芳，等 . 肺痹汤治疗特发性肺间质纤维化30例临床观察［J］. 现代中医临床， 2015 ， 22 （ 4 ）： 26 - 28 .

FU X F ， WU Z S ， CAO F ， et al . Clinical observation of Feibi Tang in the treatment of idiopathic pulmonary fibrosis in 30 cases ［J］. J Mod Clin Med ， 2015 ， 22 （ 4 ）： 26 - 28 .

LIU Z H ， LI G D ， HAO Q X ， et al . Acute exacerbation of idiopathic pulmonary fibrosis treated using the Feibi recipe：Two case reports ［J］. World J Clin Cases ， 2023 ， 11 （ 24 ）： 5742 - 5748 .

LIU H G ， PANG Q L ， CAO F ， et al . Number 2 Feibi recipe ameliorates pulmonary fibrosis by inducing autophagy through the GSK-3 β /mTOR pathway ［J］. Front Pharmacol ， 2022 ， 13 ： 921209 .

GU X F ， LONG Q ， WEI W ， et al . Number 2 Feibi recipe inhibits HO-mediated oxidative stress damage of alveolar epithelial cells by regulating the balance of mitophagy/apoptosis ［J］. Front Pharmacol ， 2022 ， 13 ： 830554 .

庞庆禄，李国栋，曹芳，等 . 肺痹汤2号对肺纤维化模型小鼠肺组织ROS/Hippo/YAP信号通路的影响［J］. 中医杂志， 2022 ， 63 （ 21 ）： 2081 - 2087 .

PANG Q L ， LI GD ， CAO F ， et al . Effect of Feibi decoction Ⅱ on ROS/Hippo/YAP signaling pathway in lung tissue of pulmonary fibrosis model mice ［J］. J Tradit Chin Med ， 2022 ， 63 （ 21 ）： 2081 - 2087 .

徐成岩，梁笑研，刘艳丽，等 . 肺痹汤2号改善放射性肺纤维化的网络药理学分析［J］. 生命的化学， 2025 ， 45 （ 8 ）： 14 .

XU C Y ， LIANG X Y ， LIU Y L ， et al . Network pharmacology-based analysis of number 2 feibi recipe for the treatment of radiation-induced pulmonary fibrosis ［J］. Chem Life ， 2025 ， 45 （ 8 ）： 14 .

MOELLER A ， ASK K ， WARBURTON D ， et al . The bleomycin animal model：A useful tool to investigate treatment options for idiopathic pulmonary fibrosis？［J］. Int J Biochem Cell Biol ， 2008 ， 40 （ 3 ）： 362 - 382 .

RAGHU G ， REMY-JARDIN M ， RICHELDI L ， et al . Idiopathic pulmonary fibrosis （an update） and progressive pulmonary fibrosis in adults：An official ATS/ERS/JRS/ALAT clinical practice guideline ［J］. Am J Respir Crit Care Med ， 2022 ， 205 （ 9 ）： e18 - e47 .

杭程，肖洋，王高雷，等 . 米烈汉教授基于宗气为本防治肺纤维化经验浅析［J］. 陕西中医， 2021 ， 42 （ 9 ）： 1282 - 1284 .

HANG C ， XIAO Y ， WANG G L ， et al . A Brief analysis of professor MI Lihan's experience in preventing and treating pulmonary fibrosis based on Zongqi as the foundation ［J］. Shaanxi J Tradit Chin Med ， 2021 ， 42 （ 9 ）： 1282 - 1284 .

杨效华，张春艳，孙海燕，等 . 周平安教授治疗特发性肺间质纤维化经验［J］. 中国临床医生， 2012 ， 40 （ 4 ）： 60 - 62 .

YANG X H ， ZHANG C Y ， SUN H Y ， et al . Professor ZHOU Ping'an's experience in treating idiopathic pulmonary interstitial fibrosis ［J］. Chin J Clin ， 2012 ， 40 （ 4 ）： 60 - 62 .

LU W ， TEOH A ， WATERS M ， et al . Pathology of idiopathic pulmonary fibrosis with particular focus on vascular endothelium and epithelial injury and their therapeutic potential ［J］. Pharmacol Ther ， 2025 ， 265 ： 108757 .

BUENO M ， CALYECA J ， ROJAS M ， et al . Mitochondria dysfunction and metabolic reprogramming as drivers of idiopathic pulmonary fibrosis ［J］. Redox Biol ， 2020 ， 33 ： 101509 .

LI J ， ZHAI X ， SUN X ， et al . Metabolic reprogramming of pulmonary fibrosis ［J］. Front Pharmacol ， 2022 ， 13 ： 1031890 .

ZHENG H ， ZHANG L ， WANG C ， et al . Metabolic dysregulation in pulmonary fibrosis：Insights into amino acid contributions and therapeutic potential ［J］. Cell Death Discov ， 2025 ， 11 （ 1 ）： 411 .

NIETHAMER T K ， STABLER C T ， LEACH J P ， et al . Defining the role of pulmonary endothelial cell heterogeneity in the response to acute lung injury ［J］. Elife ， 2020 ， 9 ： e53072 .

DESAI T J ， BROWNFIELD D G ， KRASNOW M A . Alveolar progenitor and stem cells in lung development，renewal and cancer ［J］. Nature ， 2014 ， 507 （ 7491 ）： 190 - 194 .

WANG J Y ， MICHKI S N ， SITARAMAN S ， et al . Dysregulated alveolar epithelial cell progenitor function and identity in Hermansky-Pudlak syndrome ［J］. JCI Insight ， 2024 ， 10 （ 3 ）： e183483 .

SU W ， GUO Y ， WANG Q ， et al . YAP1 inhibits the senescence of alveolar epithelial cells by targeting Prdx3 to alleviate pulmonary fibrosis ［J］. Exp Mol Med ， 2024 ， 56 （ 7 ）： 1643 - 1654 .

LIANG J ， HUANG G ， LIU X ， et al . The ZIP8/SIRT1 axis regulates alveolar progenitor cell renewal in aging and idiopathic pulmonary fibrosis ［J］. J Clin Invest ， 2022 ， 132 （ 11 ）： e157338 .

BURMAN A ， KROPSKI J A ， CALVI C L ， et al . Localized hypoxia links ER stress to lung fibrosis through induction of C/EBP homologous protein ［J］. JCI Insight ， 2018 ， 3 （ 16 ）： e99543 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Treatment of Idiopathic Pulmonary Fibrosis from Impediment

Protective Effect and Potential Mechanism of Danggui Shaoyaosan on Diabetic Kidney Disease in db/db Mice Based on Endoplasmic Reticulum Stress in Glomerular Endothelial Cells

Mechanism of Shenmai Injection to Improve Cisplatin Resistance in NSCLC Based on Endoplasmic Reticulum Stress Through PERK/ATF4/CHOP Signaling Pathway

Traditional Chinese Medicine in Regulating Endoplasmic Reticulum Stress for Treatment of Ulcerative Colitis： A Review

Related Author

CHEN Siyu

CAO Zhenghua

XU Rong

LI Qingrong

BI Yanze

SHANG Boyi

HU Shaodan

LI Ruijia

Related Institution

Changchun University of Chinese Medicine

The Affiliated Hospital of Changchun University of Chinese Medicine

Graduate School， Hebei University of Traditional Chinese Medicine

Hebei Provincial Hospital of Traditional Chinese Medicine

The Second Clinic of Hebei Provincial Directly Subordinate Organs

AI脑图

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 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