Original articles
Yunhao Wu, Xiu Yu, Yuwei Wang, Yalin Huang, Jiahui Tang, Shuaishuai Gong, Siyu Jiang, Yuanli Xia, Fang Li, Boyang Yu, Yuanyuan Zhang, Junping Kou. Ruscogenin alleviates LPS-triggered pulmonary endothelial barrier dysfunction through targeting NMMHC IIA to modulate TLR4 signaling[J]. Acta Pharmaceutica Sinica B, 2022, 12(3): 1198-1212

Ruscogenin alleviates LPS-triggered pulmonary endothelial barrier dysfunction through targeting NMMHC IIA to modulate TLR4 signaling
Yunhao Wu, Xiu Yu, Yuwei Wang, Yalin Huang, Jiahui Tang, Shuaishuai Gong, Siyu Jiang, Yuanli Xia, Fang Li, Boyang Yu, Yuanyuan Zhang, Junping Kou
State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of TCM Evaluation and Translational Research, Department of Pharmacology of Chinese Material Medica, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China
Pulmonary endothelial barrier dysfunction is a hallmark of clinical pulmonary edema and contributes to the development of acute lung injury (ALI). Here we reported that ruscogenin (RUS), an effective steroidal sapogenin of Radix Ophiopogon japonicus, attenuated lipopolysaccharides (LPS)-induced pulmonary endothelial barrier disruption through mediating non-muscle myosin heavy chain IIA (NMMHC IIA)-Toll-like receptor 4 (TLR4) interactions. By in vivo and in vitro experiments, we observed that RUS administration significantly ameliorated LPS-triggered pulmonary endothelial barrier dysfunction and ALI. Moreover, we identified that RUS directly targeted NMMHC IIA on its N-terminal and head domain by serial affinity chromatography, molecular docking, biolayer interferometry, and microscale thermophoresis analyses. Downregulation of endothelial NMMHC IIA expression in vivo and in vitro abolished the protective effect of RUS. It was also observed that NMMHC IIA was dissociated from TLR4 and then activating TLR4 downstream Src/vascular endothelial cadherin (VE-cadherin) signaling in pulmonary vascular endothelial cells after LPS treatment, which could be restored by RUS. Collectively, these findings provide pharmacological evidence showing that RUS attenuates LPS-induced pulmonary endothelial barrier dysfunction by inhibiting TLR4/Src/VE-cadherin pathway through targeting NMMHC IIA and mediating NMMHC IIA-TLR4 interactions.
Key words:    Ruscogenin    Acute lung injury    Lipopolysaccharide    Endothelial barrier    Non-muscle myosin heavy chain IIA    TLR4    VE-cadherin    Interaction   
Received: 2021-07-29     Revised: 2021-08-16
DOI: 10.1016/j.apsb.2021.09.017
Funds: The present study was supported by the National Natural Science Foundation of China (No. 81773971, China), and Double First-Class University Project (No. CPU2018GF07, China).
Corresponding author: Yuanyuan Zhang,E-mai:yuanyuanzhang@cpu.edu.cn;Junping Kou,E-mai:junpingkou@cpu.edu.cn     Email:yuanyuanzhang@cpu.edu.cn;junpingkou@cpu.edu.cn
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Yunhao Wu
Xiu Yu
Yuwei Wang
Yalin Huang
Jiahui Tang
Shuaishuai Gong
Siyu Jiang
Yuanli Xia
Fang Li
Boyang Yu
Yuanyuan Zhang
Junping Kou

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