S1P/S1PR2 axis promotes cardiac fibroblast activation and collagen secretion by blocking autophagic flux, while restoring autophagic flux reverses fibrosis

Liu Jia; Aji Mayire; Xie Yaohui; Wang Xumin

doi:10.16016/j.2097-0927.202608001

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S1P/S1PR2 axis promotes cardiac fibroblast activation and collagen secretion by blocking autophagic flux, while restoring autophagic flux reverses fibrosis

Basic Medicine | 更新时间：2026-10-30

- S1P/S1PR2 axis promotes cardiac fibroblast activation and collagen secretion by blocking autophagic flux, while restoring autophagic flux reverses fibrosis
- Journal of Army Medical University Vol. 48, Issue 20, Pages: 2908-2919(2026)
- 作者机构：
  
  新疆医科大学第六附属医院心血管内科，新疆乌鲁木齐
- 作者简介：
  
  Wang Xumin，E-mail：wangxuminsparky@163.com
- 基金信息：
  
  the General Project of Natural Science Foundation of Xinjiang Uygur Autonomous Region(2021D01C457;2024D01C146)
- DOI：10.16016/j.2097-0927.202608001
  CLC： R322.11;R329.26;R363.21
- Received：02 August 2026，
  
  Revised：2026-09-07，
  
  Accepted：12 September 2026，
  
  Published：30 October 2026
- 稿件说明：
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刘佳，玛依热·阿吉，谢耀慧，等.S1P/S1PR2信号轴通过阻断自噬流促进心肌成纤维细胞活化与胶原分泌而恢复自噬流可逆转纤维化［J］.陆军军医大学学报，2026，48（20）：2908-2919.DOI：10.16016/j.2097-0927.202608001.

Liu J，Aji M Y R，Xie Y H，et al.S1P/S1PR2 axis promotes cardiac fibroblast activation and collagen secretion by blocking autophagic flux， while restoring autophagic flux reverses fibrosis［J］.J Army Med Univ，2026，48（20）：2908-2919.DOI：10.16016/j.2097-0927.202608001.
刘佳，玛依热·阿吉，谢耀慧，等.S1P/S1PR2信号轴通过阻断自噬流促进心肌成纤维细胞活化与胶原分泌而恢复自噬流可逆转纤维化［J］.陆军军医大学学报，2026，48（20）：2908-2919.DOI：10.16016/j.2097-0927.202608001. DOI：

Liu J，Aji M Y R，Xie Y H，et al.S1P/S1PR2 axis promotes cardiac fibroblast activation and collagen secretion by blocking autophagic flux， while restoring autophagic flux reverses fibrosis［J］.J Army Med Univ，2026，48（20）：2908-2919.DOI：10.16016/j.2097-0927.202608001. DOI：

摘要

目的

心肌成纤维细胞（cardiac fibroblasts，CFs）异常活化及胶原过度沉积是心肌纤维化发生发展的关键病理环节。本研究旨在探讨1-磷酸鞘氨醇（sphingosine-1-phosphate，S1P）/1-磷酸鞘氨醇受体2（sphingosine-1-phosphate receptor 2，S1PR2）信号轴对CFs胶原分泌及纤维化表型的调控作用，并从自噬流角度阐明其分子机制。

方法

选取健康清洁级SD大鼠乳鼠3只（出生1~3 d，雌雄不限，体质量5~8 g），采用差速贴壁法分离培养大鼠原代CFs，通过波形蛋白（Vimentin）免疫荧光染色进行纯度鉴定。①探讨S1P/S1PR2信号轴对CFs活化与胶原分泌的影响，将CFs分为Control组、S1P组、S1P+S1PR2 特异性拮抗剂（JTE-013）组、S1P+小干扰RNA阴性对照（small interfering RNA negative control，si-NC）组及S1P+敲低S1PR2小干扰RNA（small interfering RNA targeting S1PR2，si-S1PR2）组，采用CCK-8法检测细胞活性，划痕实验检测细胞迁移能力，ELISA法检测细胞培养上清液中Ⅰ型胶原（collagen type Ⅰ，Col Ⅰ）及Ⅲ型胶原（collagen type Ⅲ，Col Ⅲ）含量，免疫荧光染色及Western blotting检测细胞中α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）、S1PR2及鞘氨醇激酶1（sphingosine kinase 1，SphK1）的表达。②探究自噬流在S1P/S1PR2信号轴中的调控作用，将CFs分为Control组、S1P组、S1P+JTE-013 组、S1P+自噬诱导剂雷帕霉素（rapamycin，Rapa）组和S1P+自噬流阻断剂巴弗洛霉素A1（bafilomycin A1，Baf A1）组，采用mRFP-GFP-LC3双荧光腺病毒转染观察自噬流变化，Western blotting检测自噬相关蛋白［微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3，LC3）-Ⅱ/LC3-Ⅰ、p62、组织蛋白酶D（cathepsin D，Cath D）］及α-SMA的表达水平。

结果

原代培养的大鼠CFs生长状态良好且形态典型，经Vimentin免疫荧光鉴定阳性率达95%以上，纯度符合实验要求。①与Control组比较，S1P组CFs的活性与迁移能力显著增强，Col Ⅰ、Col Ⅲ分泌量显著增加，α-SMA荧光染色强度及α-SMA、S1PR2、SphK1蛋白表达显著上调（

0.05）。与S1P组比较，S1P+JTE-013组CFs的活性与迁移能力显著降低，Col Ⅰ、Col Ⅲ分泌量显著减少，α-SMA荧光染色强度及α-SMA、S1PR2、SphK1蛋白表达显著下调（

0.05）。与S1P+si-NC组比较，S1P+si-S1PR2组CFs的活性与迁移能力显著降低，Col Ⅰ、Col Ⅲ分泌量显著减少，α-SMA荧光染色强度及α-SMA、S1PR2、SphK1蛋白表达显著下调（

0.05）。②与Control组比较，S1P组细胞内黄色荧光斑点大量积聚，LC3-Ⅱ/LC3-Ⅰ比值及p62蛋白表达显著

上调，而Cath D蛋白表达显著下调（

0.05）。与S1P组比较，S1P+JTE-013组和S1P+Rapa组红色荧光斑点数显著增加，黄色斑点数显著减少，p62蛋白表达显著下调，Cath D蛋白表达显著上调（

0.05）。此外，与S1P组比较，S1P+JTE-013组中LC3-Ⅱ/LC3-Ⅰ比值显著降低，S1P+Rapa组中LC3-Ⅱ/LC3-Ⅰ比值显著升高（

0.05）；而与S1P组比较，S1P+Baf A1组自噬流进一步受阻，表现为红色斑点数显著减少、黄色斑点数显著增加，LC3-Ⅱ/LC3-Ⅰ比值显著升高，p62和α-SMA蛋白表达显著上调，Cath D蛋白表达显著下调，且Col Ⅰ、Col Ⅲ分泌量显著增加（

0.05）。

结论

S1P/S1PR2信号轴通过阻断自噬流降解，促进心肌成纤维细胞活化与胶原分泌，而恢复自噬流可有效逆转S1P诱导的CFs纤维化表型。

Abstract

Objective

Abnormal activation of cardiac fibroblasts (CFs) and excessive collagen deposition are critical pathological events in the development of myocardial fibrosis. This study aimed to investigate the regulatory role of the sphingosine-1-phosphate (S1P)/sphingosine-1-phosphate receptor 2 (S1PR2) signaling axis in collagen secretion and the fibrotic phenotype of CFs

and to elucidate the molecular mechanisms from the perspective of autophagic flux.

Methods

Three healthy clean-grade neonatal SD rats (aged 1 to 3 d

weighing 5 to 8 g

either sex) were selected

primary rat CFs were isolated and cultured using differential adhesion method

and their purity was confirmed by Vimentin immunofluorescence staining. ① To investigate the effects of the S1P/S1PR2 signaling axis on CF activation and collagen secretion

CFs were divided into Control group

S1P group

S1P+S1PR2 specific antagonist (JTE-013) group

S1P+small interfering RNA negative control transfection (si-NC) group

and S1P+S1PR2 knockdown (si-S1PR2) group. Cell viability and migration were detected using CCK-8 assay and scratch wound assay

and the contents of type Ⅰ collagen (Col Ⅰ) and type Ⅲ collagen (Col Ⅲ) in cell culture supernatant was measured with ELISA. The expression of α-smooth muscle actin (α-SMA)

S1PR2

and sphingosine kinase 1 (SphK1) was detected by immunofluorescence staining and Western blotting. ② To explore the regulatory role of autophagic flux in the S1P/S1PR2 signaling axis

CFs were divided into Control group

S1P group

S1P + JTE-013 group

S1P + autophagy inducer rapamycin (Rapa) group

and S1P + bafilomycin A1 (Baf A1

autophagic flux inhibitor) group. Changes in autophagic flux were observed using mRFP-GFP-LC3 dual-fluorescence adenovirus transfection

and the expression levels of autophagy-related proteins [microtubule-associated protein 1 light chain 3 (LC3-Ⅱ/LC3-Ⅰ)

p62

and cathepsin D (Cath D)] and α-SMA were detected by Western blotting.

Results

Primary cultured rat CFs exhibited robust growth with typical morphology

and the positive rate of Vimentin immunofluorescence identification was over 95%

meeting the experimental purity requirement. ① Compared with the Control group

the S1P group showed significantly enhanced CF proliferation and migration abilities

increased Col Ⅰ and Col Ⅲ secretion

enhanced α-SMA fluorescence staining intensity

and upregulated protein expression levels of α-SMA

S1PR2

and SphK1 (

0.05). Compared with the S1P group

CF viability and migration capacity were significantly decreased

Col Ⅰ and Col Ⅲ secretion was significantly reduced

and α-SMA fluorescence intensity along with α-SMA

S1PR2

and SphK1 protein expression were significantly decreased in the S1P+JTE-013 group (

0.05); similarly

compared with the S1P+si-NC group

all the aforementioned parameters were significantly reduced in the S1P+si-S1PR2 group (

0.05). ② Compared with the Control group

the S1P group exhibited significant accumulation of yellow fluorescent puncta

higher LC3-Ⅱ/LC3-Ⅰ ratio

and upregulated p62 expression

while downregulated Cath D expression (

0.05). Compared with the S1P group

the S1P+JTE-013 and S1P+Rapa groups showed significantly increased number of red fluorescent puncta

decreased number of yellow puncta

downregulated p62 and upregulated Cath D expression (

0.05). Additionally

compared with the S1P group

the LC3-Ⅱ/LC3-Ⅰ ratio was significantly decreased in the S1P+JTE-013 group

whereas increased in the S1P+Rapa group (

0.05). In contrast

compared with the S1P group

the S1P+Baf A1 group showed further blocked autophagic flux

manifested as significantly reduced number of red puncta and increased number of yellow puncta

increased LC3-Ⅱ/LC3-Ⅰ ratio

upregulated expression of p62 and α-SMA while downregulated Cath D

and enhanced secretion of Col Ⅰ and Col Ⅲ (

0.05).

Conclusion

The S1P/S1PR2 signaling axis promotes cardiac fibroblast activation and collagen secretion by blocking autophagic flux degradation

and restoring autophagic flux can effectively reverse the S1P-induced fibrotic phenotype in CFs.

关键词

Keywords

references

Chimenti I ， Pagano F ， Cozzolino C ， et al . The role of cardiac fibroblast heterogeneity in myocardial fibrosis and its novel therapeutic potential ［J］. Int J Mol Sci ， 2025 ， 26 （ 12 ）： 5882 . DOI： 10.3390/ijms26125882 http://dx.doi.org/10.3390/ijms26125882 .

Song M ， Choi D B ， Im J S ， et al . Modeling acute myocardial infarction and cardiac fibrosis using human induced pluripotent stem cell-derived multi-cellular heart organoids ［J］. Cell Death Dis ， 2024 ， 15 （ 5 ）： 308 . DOI： 10.1038/s41419-024-06703-9 http://dx.doi.org/10.1038/s41419-024-06703-9 .

Ghazal R ， Wang M ， Liu D ， et al . Cardiac fibrosis in the multi-omics era： implications for heart failure ［J］. Circ Res ， 2025 ， 136 （ 7 ）： 773 - 802 . DOI： 10.1161/CIRCRESAHA. 124. 325402 http://dx.doi.org/10.1161/CIRCRESAHA.124.325402 .

Benkhoff M ， Polzin A . Lipoprotection in cardiovascular diseases ［J］. Pharmacol Ther ， 2024 ， 264 ： 108747 . DOI： 10.1016/j.pharmthera.2024.108747 http://dx.doi.org/10.1016/j.pharmthera.2024.108747 .

Wang N ， Li J Y ， Zeng B ， et al . Sphingosine-1-phosphate signaling in cardiovascular diseases ［J］. Biomolecules ， 2023 ， 13 （ 5 ）： 818 . DOI： 10.3390/biom13050818 http://dx.doi.org/10.3390/biom13050818 .

Zhao M ， Bian R T ， Xu X G ， et al . Sphingolipid metabolism and signalling pathways in heart failure： from molecular mechanism to therapeutic potential ［J］. J Inflamm Res ， 2025 ， 18 ： 5477 - 5498 . DOI： 10.2147/JIR.S515757 http://dx.doi.org/10.2147/JIR.S515757 .

Donati C ， Cencetti F ， Bernacchioni C ， et al . Role of sphingosine 1-phosphate signalling in tissue fibrosis ［J］. Cell Signal ， 2021 ， 78 ： 109861 . DOI： 10.1016/j.cellsig. 2020. 109861 http://dx.doi.org/10.1016/j.cellsig.2020.109861 .

Ouyang J ， Long H J ， Chen S H ， et al . S1PR2 mediates smooth muscle cell proliferation and endothelial cell permeability via Akt/mTOR and RhoA/ROCK1 in atherosclerosis ［J］. J Biochem Mol Toxicol ， 2025 ， 39 （ 6 ）： e70351 . DOI： 10.1002/jbt.70351 http://dx.doi.org/10.1002/jbt.70351 .

Sawai T ， Nakamura Y ， Arawaka S . Secretory autophagy mediates lysosomal and autophagic degradation for α-synuclein proteostasis ［J］. J Biol Chem ， 2025 ， 301 （ 8 ）： 110474 . DOI： 10.1016/j.jbc.2025.110474 http://dx.doi.org/10.1016/j.jbc.2025.110474 .

余辉，王先敏，藏登，等 . 基于m6A激活自噬探讨糖肾灌肠方对糖尿病肾病肾纤维化的改善作用［J］. 中成药， 2025 ， 47 （ 10 ）： 3473 - 3479 . DOI： 10.3969/j.issn.1001-1528.2025.10.045 http://dx.doi.org/10.3969/j.issn.1001-1528.2025.10.045 .

Yu H ， Wang X M ， Zang D ， et al . Based on M6A-activated autophagy， the improvement effect of Tangshen Enema recipe on renal fibrosis in diabetic nephropathy was discussed ［J］. Chin Tradit Pat Med ， 2025 ， 47 （ 10 ）： 3473 - 3479 . DOI： 10.3969/j.issn.1001-1528.2025.10.045 http://dx.doi.org/10.3969/j.issn.1001-1528.2025.10.045 .

Assi M ， Wang R H ， Kawaler E A ， et al . Extracellular matrix sensing regulates intratumoral heterogeneity of autophagic flux ［J］. Cell ， 2026 ， 189 （ 6 ）： 1731 - 1747.e25 . DOI： 10.1016/j.cell.2025.12.053 http://dx.doi.org/10.1016/j.cell.2025.12.053 .

Li Y Y ， Meng F L ， Zhou S Y ， et al . CDX1 improves nicotine induced cardiac fibroblasts activation and cardiomyocyte hypertrophy by alleviating autophagic flux impairment through modulation of LAPTM4B ［J］. Sci Rep ， 2025 ， 15 （ 1 ）： 9985 . DOI： 10.1038/s41598-025-94160-5 http://dx.doi.org/10.1038/s41598-025-94160-5 .

Song C P ， Wang S W ， Guo X L ， et al . Myocardial bridging in obstructive hypertrophic cardiomyopathy： a risk factor for myocardial fibrosis ［J］. BMC Med ， 2024 ， 22 （ 1 ）： 86 . DOI： 10.1186/s12916-024-03301-6 http://dx.doi.org/10.1186/s12916-024-03301-6 .

Feng Y T ， Chen Z S ， Gao Y H ， et al . CKAP4 promotes atrial fibrosis and enhances atrial fibrillation vulnerability via WNT/β-catenin activation ［J］. Circ Arrhythm Electrophysiol ， 2025 ， 18 （ 11 ）： e014217 . DOI： 10.1161/CIRCEP.125.014217 http://dx.doi.org/10.1161/CIRCEP.125.014217 .

Kleinbongard P ， Senyo S E ， Lindsey M L ， et al . Cardiac fibroblasts： answering the call ［J］. Am J Physiol Heart Circ Physiol ， 2024 ， 327 （ 3 ）： H681 - H686 . DOI： 10.1152/ajpheart.00478.2024 http://dx.doi.org/10.1152/ajpheart.00478.2024 .

El-Maadawy W H ， Amer M ， Mostafa A ， et al . Ticagrelor attenuates cholestasis-induced liver fibrosis by inhibiting S1PR2-dependent Akt/ERK signaling， NLRP3 inflammasome activation ［J］. Int Immunopharmacol ， 2026 ， 168 ： 115834 . DOI： 10.1016/j.intimp.2025.115834 http://dx.doi.org/10.1016/j.intimp.2025.115834 .

Park S ， Kim J ， Yang S ， et al . Exogenous S1P via S1P receptor 2 induces CTGF expression through Src-RhoA-ROCK-YAP pathway in hepatic stellate cells ［J］. Mol Biol Rep ， 2024 ， 51 （ 1 ）： 950 . DOI： 10.1007/s11033-024-09868-w http://dx.doi.org/10.1007/s11033-024-09868-w .

Wang X M ， Liu C ， Li X R ， et al . JTE-013 reduces inflammation and improves pulmonary fibrosis by regulating the TRAF2/NF-κB/IGF-1 signaling pathway and macrophage polarization ［J］. Int Immunopharmacol ， 2025 ， 167 ： 115727 . DOI： 10.1016/j.intimp.2025.115727 http://dx.doi.org/10.1016/j.intimp.2025.115727 .

Venugopal H ， Hanna A ， Humeres C ， et al . Properties and functions of fibroblasts and myofibroblasts in myocardial infarction ［J］. Cells ， 2022 ， 11 （ 9 ）： 1386 . DOI： 10.3390/cells11091386 http://dx.doi.org/10.3390/cells11091386 .

Pitman M R ， Lewis A C ， Davies L T ， et al . The sphingosine 1-phosphate receptor 2/4 antagonist JTE-013 elicits off-target effects on sphingolipid metabolism ［J］. Sci Rep ， 2022 ， 12 （ 1 ）： 454 . DOI： 10.1038/s41598-021-04009-w http://dx.doi.org/10.1038/s41598-021-04009-w .

李辉，孟丽红，薛晓明，等 . 基于自噬探讨养阴益气方干预博来霉素诱导的大鼠特发性肺纤维化作用机制［J］. 中国中医药信息杂志， 2026 ， 33 （ 1 ）： 64 - 69 . DOI： 10.19879/j.cnki.1005-5304.202506411 http://dx.doi.org/10.19879/j.cnki.1005-5304.202506411 .

Li H ， Meng L H ， Xue X M ， et al . Exploration on the mechanism of Yangyin Yiqi prescription in bleomycin-induced idiopathic pulmonary fibrosis in rats based on autophagy ［J］. Chin J Inf Tradit Chin Med ， 2026 ， 33 （ 1 ）： 64 - 69 . DOI： 10.19879/j.cnki.1005-5304.202506411 http://dx.doi.org/10.19879/j.cnki.1005-5304.202506411 .

Wang C X ， Zeng X J ， Yang P F ， et al . Augmenter of liver regeneration inhibits renal fibrosis during acute kidney injury to chronic kidney disease transition by regulating autophagic flux ［J］. Arch Biochem Biophys ， 2025 ， 764 ： 110218 . DOI： 10.1016/j.abb.2024.110218 http://dx.doi.org/10.1016/j.abb.2024.110218 .

赵晓彬，靳文芳，张海峰，等 . 线粒体质量控制在心肌纤维化中的研究进展［J］. 中国药理学通报， 2025 ， 41 （ 8 ）： 1419 - 1423 . DOI： 10.12360/CPB202408079 http://dx.doi.org/10.12360/CPB202408079 .

Zhao X B ， Jin W F ， Zhang H F ， et al . Research progress of mitochondrial quality control in myocardial fibrosis ［J］. Chin Pharmacol Bull ， 2025 ， 41 （ 8 ）： 1419 - 1423 . DOI： 10.12360/CPB202408079 http://dx.doi.org/10.12360/CPB202408079 .

Sun X X ， Qi L ， Dai M J ， et al . Suppression of OTUD4 protects against myocardial ischemia-reperfusion injury by increasing autophagic flux and inhibiting apoptosis in cardiomyocytes ［J］. J Mol Med ， 2025 ， 103 （ 11 ）： 1567 - 1582 . DOI： 10.1007/s00109-025-02605-1 http://dx.doi.org/10.1007/s00109-025-02605-1 .

Li Y ， Feng C N ， Zu Y G ， et al . VMP1 enhances autophagy to mitigate cardiac fibroblast activation and reduce post-ischemia-reperfusion fibrosis ［J］. Cell Signal ， 2025 ， 136 ： 112105 . DOI： 10.1016/j.cellsig.2025.112105 http://dx.doi.org/10.1016/j.cellsig.2025.112105 .

Ito S I ， Tanaka Y . Evaluation of LC3-Ⅱ release via extracellular vesicles in relation to the accumulation of intracellular LC3-positive vesicles ［J］. JoVE ， 2024 （ 212 ）： e67385 . DOI： 10.3791/67385 http://dx.doi.org/10.3791/67385 .

Lee S J ， Kim H Y ， Lee M J ， et al . Characterization and chemical modulation of p62/SQSTM1/Sequestosome-1 as an autophagic N-recognin ［J］. Methods Enzymol ， 2023 ： 235 - 265 . DOI： 10.1016/bs.mie.2023.02.005 http://dx.doi.org/10.1016/bs.mie.2023.02.005 .

Mijanovic O ， Petushkova A I ， Brankovic A ， et al . Cathepsin D-managing the delicate balance ［J］. Pharmaceutics ， 2021 ， 13 （ 6 ）： 837 . DOI： 10.3390/pharmaceutics13060837 http://dx.doi.org/10.3390/pharmaceutics13060837 .

Cheng A F ， Tse K H ， Chow H M ， et al . ATM loss disrupts the autophagy-lysosomal pathway ［J］. Autophagy ， 2021 ， 17 （ 8 ）： 1998 - 2010 . DOI： 10.1080/15548627.2020.1805860 http://dx.doi.org/10.1080/15548627.2020.1805860 .

Bailus B J ， Scheeler S M ， Simons J ， et al . Modulating FKBP5/FKBP51 and autophagy lowers HTT （huntingtin） levels ［J］. Autophagy ， 2021 ， 17 （ 12 ）： 4119 - 4140 . DOI： 10.1080/15548627.2021.1904489 http://dx.doi.org/10.1080/15548627.2021.1904489 .

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