BACKGROUND AND AIM: Zhilong Huoxue Tongyu (ZL) capsule is a classical traditional Chinese medicine (TCM) with satisfactory curative effects. Endothelial cell (EC) dysfunction plays an important role during myocardial fibrosis (MF). But the therapeutic effect of ZL capsule on EC dysfunction remains unknown in the development of MF. This study aims to investigate the effect of ZL capsule on EC dysfunction during MF in vivo. EXPERIMENTAL PROCEDURE: The model of MF is established in vivo by injecting isoproterenol for 14 days, simultaneously, we examined the therapeutic effect of ZL capsule on MF in vivo. An integrative approach combining biomarker examination, echocardiography and myocardial fibrosis condition using Hematoxylin-eosin staining, Masson staining, and Sirius red staining were performed to assess the efficacy of ZL capsule against MF. Subsequently, comprehensive immunofluorescence staining was performed to evaluate the therapeutic effect of ZL capsule on EC dysfunction. RESULTS AND CONCLUSION: Prior to experiments, analysis of the published single-cell sequencing data was performed and it was discovered that EC dysfunction plays an important role. Further pharmacological results showed that ZL capsule could alleviate fibrosis injury and collagen fiber deposition. The mechanism investigation results showed that the endothelial-to-mesenchymal transition (EndMT) and MHC class-II (MHC-II) expression in EC were improved. In addition, ZL capsule can attenuate the inflammatory response during MF by intervening the activation of CD4+T cell mediated by EC. For the first time, we provided evidence that ZL capsule could improve MF by alleviating EC dysfunction via the regulation of EndMT and expression of MHC-II. TAXONOMY CLASSIFICATION BY EVISE: Myocardial fibrosis, Chinese Herbal Medicine, Traditional Medicine, Endothelium, dysfunction, Endothelial-to-mesenchymal transition.

Department of Cardiovascular Medicine The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University Luzhou Sichuan China

Institute of Integrated Chinese and Western Medicine Southwest Medical University Luzhou 646000 China

National Traditional Chinese Medicine Clinical Research Base and Drug Research Center of the Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University Luzhou 646000 China

State Key Laboratories for Quality Research in Chinese Medicines Faculty of Chinese Medicine Macau University of Science and Technology Macau 853 China

The Czech Center for Traditional Chinese Medicine Jeremenkova 1211 40 Olomouc 77900 Czech Republic

The National T C M Service Export Base of the Affiliated T C M Hospital of Southwest Medical University Luzhou 646000 China

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Yan F., Sun Y., Mao Y., et al. Ultrasound molecular imaging of atherosclerosis for early diagnosis and therapeutic evaluation through leucocyte-like multiple targeted microbubbles. Theranostics. 2018;8(7):1879–1891. PubMed PMC

Roth G.A., Mensah G.A., Johnson C.O., et al. Global burden of cardiovascular diseases and risk factors, 1990-2019: update from the GBD 2019 study. J Am Coll Cardiol. 2020;76(25):2982–3021. PubMed PMC

Aimo A., Gaggin H.K., Barison A., Emdin M., Januzzi J.L., Jr. Imaging, biomarker, and clinical predictors of cardiac remodeling in heart failure with reduced ejection fraction. JACC Heart Fail. 2019;7(9):782–794. PubMed

Zhu Z.D., Ye J.M., Fu X.M., et al. DDAH2 alleviates myocardial fibrosis in diabetic cardiomyopathy through activation of the DDAH/ADMA/NOS/NO pathway in rats. Int J Mol Med. 2019;43(2):749–760. PubMed PMC

Zong Y., Huang Y., Chen S., et al. Downregulation of endogenous hydrogen sulfide pathway is involved in mitochondrion-related endothelial cell apoptosis induced by high salt. Oxid Med Cell Longev. 2015;2015 PubMed PMC

Siasos G., Sara J.D., Zaromytidou M., et al. Local low shear stress and endothelial dysfunction in patients with nonobstructive coronary atherosclerosis. J Am Coll Cardiol. 2018;71(19):2092–2102. PubMed

Eirin A., Ebrahimi B., Kwon S.H., et al. Restoration of mitochondrial cardiolipin attenuates cardiac damage in swine renovascular hypertension. J Am Heart Assoc. 2016;5(6) PubMed PMC

Morganti M., Carpi A., Nicolini A., et al. Atherosclerosis and cancer: common pathways on the vascular endothelium. Biomed Pharmacother. 2002;56(7):317–324. PubMed

Nagai N., Ohguchi H., Nakaki R., et al. Downregulation of ERG and FLI1 expression in endothelial cells triggers endothelial-to-mesenchymal transition. PLoS Genet. 2018;14(11) PubMed PMC

Wang J., Yu M., Xu J., et al. Glucagon-like peptide-1 (GLP-1) mediates the protective effects of dipeptidyl peptidase IV inhibition on pulmonary hypertension. J Biomed Sci. 2019;26(1):6. PubMed PMC

Lovisa S., Fletcher-Sananikone E., Sugimoto H., et al. Endothelial-to-mesenchymal transition compromises vascular integrity to induce Myc-mediated metabolic reprogramming in kidney fibrosis. Sci Signal. 2020;13(635) PubMed PMC

Wang B., Ge Z., Wu Y., et al. MFGE8 is down-regulated in cardiac fibrosis and attenuates endothelial-mesenchymal transition through Smad2/3-Snail signalling pathway. J Cell Mol Med. 2020;24(21):12799–12812. PubMed PMC

Carambia A., Frenzel C., Bruns O.T., et al. Inhibition of inflammatory CD4 T cell activity by murine liver sinusoidal endothelial cells. J Hepatol. 2013;58(1):112–118. PubMed

Zhang Y., Yang X., Bi T., et al. Targeted inhibition of the immunoproteasome blocks endothelial MHC class II antigen presentation to CD4(+) T cells in chronic liver injury. Int Immunopharm. 2022;107 PubMed

Weirather J., Hofmann U.D., Beyersdorf N., et al. Foxp3+ CD4+ T cells improve healing after myocardial infarction by modulating monocyte/macrophage differentiation. Circ Res. 2014;115(1):55–67. PubMed

Lu M., Qin X., Yao J., Yang Y., Zhao M., Sun L. Th17/Treg imbalance modulates rat myocardial fibrosis and heart failure by regulating LOX expression. Acta Physiol. 2020;230(3) PubMed

Zarak-Crnkovic M., Kania G., Jazwa-Kusior A., et al. Heart non-specific effector CD4(+) T cells protect from postinflammatory fibrosis and cardiac dysfunction in experimental autoimmune myocarditis. Basic Res Cardiol. 2019;115(1):6. PubMed PMC

Savvatis K., Pappritz K., Becher P.M., et al. Interleukin-23 deficiency leads to impaired wound healing and adverse prognosis after myocardial infarction. Circ Heart Fail. 2014;7(1):161–171. PubMed

Kinzel S., Lehmann-Horn K., Torke S., et al. Myelin-reactive antibodies initiate T cell-mediated CNS autoimmune disease by opsonization of endogenous antigen. Acta Neuropathol. 2016;132(1):43–58. PubMed PMC

Li H., Hong S., Qian J., Zheng Y., Yang J., Yi Q. Cross talk between the bone and immune systems: osteoclasts function as antigen-presenting cells and activate CD4+ and CD8+ T cells. Blood. 2010;116(2):210–217. PubMed PMC

Gkountidi A.O., Garnier L., Dubrot J., et al. MHC class II antigen presentation by lymphatic endothelial cells in tumors promotes intratumoral regulatory T cell-suppressive functions. Cancer Immunol Res. 2021;9(7):748–764. PubMed PMC

Liang P., Mao L., Ma Y., Ren W., Yang S. A systematic review on Zhilong Huoxue Tongyu capsule in treating cardiovascular and cerebrovascular diseases: pharmacological actions, molecular mechanisms and clinical outcomes. J Ethnopharmacol. 2021;277 PubMed

Li N., Sun J., Chen J.L., Bai X., Wang T.H. Gene network mechanism of Zhilong Huoxue Tongyu capsule in treating cerebral ischemia-reperfusion. Front Pharmacol. 2022;13 PubMed PMC

Liu M., Pu Y., Gu J., et al. Evaluation of Zhilong Huoxue Tongyu capsule in the treatment of acute cerebral infarction: a systematic review and meta-analysis of randomized controlled trials. Phytomedicine. 2021;86 PubMed

Mazhar M., Yang G., Mao L., et al. Zhilong Huoxue Tongyu capsules ameliorate early brain inflammatory injury induced by intracerebral hemorrhage via inhibition of canonical NFsmall ka, cyrillicbeta signalling pathway. Front Pharmacol. 2022;13 PubMed PMC

Chen B., Tie M.H., Wei D.X., Chen W.J., Gong R.Y. To explore the etiology and pathogenesis of vascular endothelial dysfunction of hypertension based on the theory of "Yin and Yang imbalance". Chin. J. Integr. Med. Cardio Cerebrovasc. Dis. 2021;19(13):2281–2284.

Bacmeister L., Schwarzl M., Warnke S., et al. Inflammation and fibrosis in murine models of heart failure. Basic Res Cardiol. 2019;114(3):19. PubMed

Prabhu S.D., Frangogiannis N.G. The biological basis for cardiac repair after myocardial infarction: from inflammation to fibrosis. Circ Res. 2016;119(1):91–112. PubMed PMC

Ren W., Chen S., Li S., et al. Photoluminescence enhancement of carbon dots by surfactants at room temperature. Chemistry. 2018;24(59):15806–15811. PubMed

Ren W., Han L., Luo M., et al. Multi-component identification and target cell-based screening of potential bioactive compounds in toad venom by UPLC coupled with high-resolution LTQ-Orbitrap MS and high-sensitivity Qtrap MS. Anal Bioanal Chem. 2018;410(18):4419–4435. PubMed

Liang P., Ma Y., Yang L., et al. Uncovering the mechanisms of active components from toad venom against hepatocellular carcinoma using untargeted metabolomics. Molecules. 2022;27(22) PubMed PMC

Sun T.L., Li W.Q., Tong X.L., Liu X.Y., Zhou W.H. Xanthohumol attenuates isoprenaline-induced cardiac hypertrophy and fibrosis through regulating PTEN/AKT/mTOR pathway. Eur J Pharmacol. 2021;891 PubMed

Zhao Y., Jiang Y., Chen Y., et al. Dissection of mechanisms of Chinese medicinal formula Si-Miao-Yong-an decoction protects against cardiac hypertrophy and fibrosis in isoprenaline-induced heart failure. J Ethnopharmacol. 2020;248 PubMed

Qing J., Ren Y., Zhang Y., et al. Dopamine receptor D2 antagonism normalizes profibrotic macrophage-endothelial crosstalk in non-alcoholic steatohepatitis. J Hepatol. 2022;76(2):394–406. PubMed

Chaiwong S., Chatturong U., Chanasong R., et al. Dried mulberry fruit ameliorates cardiovascular and liver histopathological changes in high-fat diet-induced hyperlipidemic mice. J Tradit Complement Med. 2021;11(4):356–368. PubMed PMC

Zeng H., Pan T., Zhan M., et al. Suppression of PFKFB3-driven glycolysis restrains endothelial-to-mesenchymal transition and fibrotic response. Signal Transduct Targeted Ther. 2022;7(1):303. PubMed PMC

Lee S., Lee D.H., Park B.W., et al. In vivo transduction of ETV2 improves cardiac function and induces vascular regeneration following myocardial infarction. Exp Mol Med. 2019;51(2):1–14. PubMed PMC

Wang Z.N., Cui M., Shah A.M., et al. Cell-type-specific gene regulatory networks underlying murine neonatal heart regeneration at single-cell resolution. Cell Rep. 2020;33(10) PubMed PMC

Chen X., Tang Y.J., Gao M., Qin S.G., Zhou J.Z., Li X.H. Prenatal exposure to lipopolysaccharide results in myocardial fibrosis in rat offspring. Int J Mol Sci. 2015;16(5):10986–10996. PubMed PMC

Li Q., Ouyang X.Y., Lin J. The impact of periodontitis on vascular endothelial dysfunction. Front Cell Infect Microbiol. 2022;12 PubMed PMC

Meng G.L., Zhu J.B., Xiao Y.J., et al. Hydrogen sulfide donor GYY4137 protects against myocardial fibrosis. Oxid Med Cell Longev. 2015;2015 PubMed PMC

Wilhelmi T., Xu X.B., Tan X.Y., et al. Serelaxin alleviates cardiac fibrosis through inhibiting endothelial-to-mesenchymal transition via RXFP1. Theranostics. 2020;10(9):3905–3924. PubMed PMC

Lang C.I., Döring P., Gäbel R., et al. [(68)Ga]-NODAGA-RGD positron emission tomography (PET) for assessment of post myocardial infarction angiogenesis as a predictor for left ventricular remodeling in mice after cardiac stem cell therapy. Cells. 2020;9(6):1358. PubMed PMC

Testai L., Brancaleone V., Flori L., Montanaro R., Calderone V. Modulation of EndMT by hydrogen sulfide in the prevention of cardiovascular fibrosis. Antioxidants. 2021;10(6):910. PubMed PMC

Chen Y., Pu Q., Ma Y., et al. Aging reprograms the hematopoietic-vascular niche to impede regeneration and promote fibrosis. Cell Metabol. 2021;33(2):395–410 e394. PubMed

van Balkom B.W.M., Eisele A.S., Pegtel D.M., Bervoets S., Verhaar M.C. Quantitative and qualitative analysis of small RNAs in human endothelial cells and exosomes provides insights into localized RNA processing, degradation and sorting. J Extracell Vesicles. 2015;4 PubMed PMC

Childs B.G., Durik M., Baker D.J., van Deursen J.M. Cellular senescence in aging and age-related disease: from mechanisms to therapy. Nat Med. 2015;21(12):1424–1435. PubMed PMC