The components of Huang-Lian-Jie-Du-Decoction act synergistically to exert protective effects in a rat ischemic stroke model

// Qian Zhang 1 , Junsong Wang 2 , Chao Zhang 1 , Shanting Liao 1 , Pei Li 1 , Dingqiao Xu 1 , Yan Lv 1 , Minghua Yang 1 , Lingyi Kong 1 1 State Key Laboratory of Natural Medicines, Department of Natural Medicinal Chemistry, China Pharmaceutical University, Nanjing, 210009, P.R. China 2 Center for Molecular Metabolism, Nanjing University of Science and Technology, Nanjing, 210094, P.R. China Correspondence to: Lingyi Kong, email: cpu_lykong@126.com Junsong Wang, email: wang.junsong@gmail.com Keywords: ischemic stroke, metabolomics, Huang-Lian-Jie-Du-Decoction, middle cerebral artery occlusion, oxidative stress Received: June 29, 2016     Accepted: September 28, 2016     Published: October 13, 2016 ABSTRACT Huang-Lian-Jie-Du-Decoction (HLJDD, Oren-gedoku-to in Japanese) is commonly used in traditional Chinese medicine (TCM) to treat ischemic stroke. This study investigated the efficacy of various combinations of the major components of HLJDD, berberine (A), baicalin (B), and jasminoidin (C), on the treatment of ischemic stroke modeled by middle cerebral artery occlusion (MCAO) in rats. The effects of A, B and C individually and their combinations were investigated using proton nuclear magnetic resonance ( 1 H NMR)-based metabolomics complemented with neurologic deficit scoring, infarct volume measurement, biochemistry, histopathology and immunohistochemistry, as well as quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Ischemic stroke produces severe oxidative stress, which induces further damage. Our results show that the ABC combination treatment increased levels of cellular antioxidants that scavenged reactive oxygen species during ischemia-reperfusion via the nuclear erythroid 2-related factor 2 (Nrf2) signaling cascade. These protective effects were not observed with the other treatments. These results suggest that a combination of component herbs in HLJDD exhibit stronger effects than the individual herbs alone. Our integrated metabolomics approach also provides a tractable, powerful tool for understanding the science behind TCM formulations.