An Integrative Pharmacology-Based Analysis of Refined Qingkailing Injection Against Cerebral Ischemic Stroke: A Novel Combination of Baicalin, Geniposide, Cholic Acid, and Hyodeoxycholic Acid.

Chongyang Ma,Jie Mu,Xueqian Wang,Tian Xu,Shuling Liu,Changxiang Li,Qingguo Wang,Changming Zhai,Shuang Zhang,Fafeng Cheng,Zisong Wang

doi:10.3389/fphar.2020.00519

Abstract

Stroke is the second leading cause of death after heart disease globally and cerebral ischemic stroke accounts for approximately 70% of all incident stroke cases. We selected four main compounds from a patent Chinese medicine, Qingkailing (QKL) injection, including baicalin from Scutellaria baicalensis Georgi (Huang Qin), geniposide from Gardenia jasminoides J. Ellis (Zhizi), and cholic acid and hyodeoxycholic acid from Bovis Calculus (Niuhuang) with a ratio of 4.4:0.4:3:2.6 m/m, to develop a more efficacious and safer modern Chinese medicine injection against ischemic stroke, refined QKL (RQKL). In this study, we investigated multiple targets, levels, and pathways of RQKL by using an integrative pharm\\acology combining experimental validation approach. In silica study showed that RQKL may regulate PI3K-Akt, estrogen, neurotrophin, HIF-1, MAPK, Hippo, FoxO, TGF-beta, NOD-like receptor, apoptosis, NF-kappa B, Wnt, chemokine, TNF, Toll-like receptor signaling pathways against ischemic stroke. The experimental results showed that RQKL improved neurological function and prevented infract volume and blood-brain-barrier damage. RQKL inhibited microgliosis and astrogliosis, and protected neurons from ischemic/reperfusion injury. RQKL also inhibited cell apoptosis and affecting the ratio of the anti-apoptosis protein B-cell lymphoma-2 (Bcl2) and pro-apoptosis protein Bcl2-associated X protein (Bax). Western blot analysis showed that RQKL activated AKT/PI3K signaling pathway and antibody array showed RQKL inhibited inflammatory response and decreased proinflammatory factor Tnf, Il6, and Il1b, and chemokines Ccl2, Cxcl2, and Cxcl3, and increased anti-inflammatory cytokine Il10. In conclusion, RQKL protected tissue against ischemic stroke through multiple-target, multiple signals, and modulating multiple cell-types in brain. This study not only promoted our understanding of the role of RQKL against ischemic stroke, but also provided a pattern for the study of Chinese medicine combining pharmaceutical Informatics and system biology methods.

Highlights

Stroke is the second leading cause of death after heart disease globally and is associated with the highest disability-adjusted lifeyears lost of any disease in China (Wu et al, 2019)
Construction and Analysis of Compound-Target Network of Refined Qingkailing RQKL is composed of baicalin, geniposide, cholic acid, and hyodeoxycholic acid, which have all been identified to be crucial bioactive natural compounds of a famous patent Chinese medicine, QKL injection
We found that 16 differential genes were overlapping among all the targets of each compound, indicating a synergistic effect of RQKL compounds on these targets, such as the dopamine receptor D2 (DRD2) and estrogen receptor 1 (ESR1), adenosine A1 receptor (ADORA1), and retinoid X receptor alpha (RXRA)

Summary

Introduction

Stroke is the second leading cause of death after heart disease globally and is associated with the highest disability-adjusted lifeyears lost of any disease in China (Wu et al, 2019). 795,000 new stroke cases are reported in the US yearly and 4.3 times this number was recently surveyed by the nationwide community-based study, NESS-China (Wang W. et al, 2017). Cerebral ischemic stroke accounts for approximately 70% of all incident stroke cases. Since the 1990s, intravenous alteplase has been considered the only evidencebased therapeutic agent for improving the prognosis of patients with cerebral ischemic stroke and accepted as standard of care all around the world (Powers et al, 2019). Exploring novel therapeutic strategies to treat ischemic stroke is an urgent need

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