Endogenous hydrogen sulfide persulfidates endothelin type A receptor to inhibit pulmonary arterial smooth muscle cell proliferation
Yanan Zhang,
Xiaoyu Tian,
Liangyi Chen,
Shiqun Zhao,
Xinjing Tang,
Xin Liu,
Dan Zhou,
Chaoshu Tang,
Bin Geng,
Junbao Du,
Hongfang Jin,
Yaqian Huang
Affiliations
Yanan Zhang
Department of Pediatrics, Peking University First Hospital, Beijing, 100034, PR China
Xiaoyu Tian
Department of Pediatrics, Peking University First Hospital, Beijing, 100034, PR China
Liangyi Chen
State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing, 100871, PR China; National Center for Nanoscience and Technology, Beijing, 100871, PR China
Shiqun Zhao
State Key Laboratory of Membrane Biology, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing, 100871, PR China; National Center for Nanoscience and Technology, Beijing, 100871, PR China
Xinjing Tang
State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, No. 38, Xueyuan Rd, Beijing, 100191, PR China
Xin Liu
Department of Pediatrics, Peking University First Hospital, Beijing, 100034, PR China
Dan Zhou
Department of Cardiology, Wuhan Children's Hospital, Wuhan, PR China
Chaoshu Tang
Department of Physiology and Pathophysiology, Peking University Health Science Center, Beijing, 100191, PR China; State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing, 100191, PR China
Bin Geng
Hypertension Center, Fuwai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100037, PR China
Junbao Du
Department of Pediatrics, Peking University First Hospital, Beijing, 100034, PR China; State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing, 100191, PR China
Hongfang Jin
Department of Pediatrics, Peking University First Hospital, Beijing, 100034, PR China; Corresponding author. Department of Pediatrics, Peking University First Hospital, No. 1 Xi-an men Street, West District, Beijing, 100034, PR China.
Yaqian Huang
Department of Pediatrics, Peking University First Hospital, Beijing, 100034, PR China; Corresponding author. Department of Pediatrics, Peking University First Hospital, No. 1 Xi-an men Street, West District, Beijing, 100034, PR China.
Background: The binding of endothelin-1 (ET-1) to endothelin type A receptor (ETAR) performs a critical action in pulmonary arterial smooth muscle cell (PASMC) proliferation leading to pulmonary vascular structural remodeling. More evidence showed that cystathionine γ-lyase (CSE)-catalyzed endogenous hydrogen sulfide (H2S) was involved in the pathogenesis of cardiovascular diseases. In this study, we aimed to explore the effect of endogenous H2S/CSE pathway on the ET-1/ETAR binding and its underlying mechanisms in the cellular and animal models of PASMC proliferation. Methods and results: Both live cell imaging and ligand-receptor assays revealed that H2S donor, NaHS, inhibited the binding of ET-1/ETAR in human PASMCs (HPASMCs) and HEK-293A cells, along with an inhibition of ET-1-activated HPASMC proliferation. While, an upregulated Ki-67 expression by the pulmonary arteries, a marked pulmonary artery structural remodeling, and an increased pulmonary artery pressure were observed in CSE knockout (CSE-KO) mice with a deficient H2S/CSE pathway compared with those in the wild type (WT) mice. Meanwhile, NaHS rescued the enhanced binding of ET-1 with ETAR and cell proliferation in the CSE-knockdowned HPASMCs. Moreover, the ETAR antagonist BQ123 blocked the enhanced proliferation of CSE-knockdowned HPASMCs. Mechanistically, ETAR persulfidation was reduced in the lung tissues of CSE-KO mice compared to that in WT mice, which could be reversed by NaHS treatment. Similarly, NaHS persulfidated ETAR in HPASMCs and HEK-293A cells. Whereas a thiol reductant dithiothreitol (DTT) reversed the H2S-induced ETAR persulfidation and further blocked the H2S-inhibited binding of ET-1/ETAR and HPASMC proliferation. Furthermore, the mutation of ETAR at cysteine (Cys) 69 abolished the persulfidation of ETAR by H2S, and subsequently blocked the H2S-suppressed ET-1/ETAR binding and HPASMC proliferation. Conclusion: Endogenous H2S persulfidated ETAR at Cys69 to inhibit the binding of ET-1 to ETAR, subsequently suppressed PASMC proliferation, and antagonized pulmonary vascular structural remodeling.
