Understanding the Mechanistic Contribution of Herbal Extracts in Compound Kushen Injection With Transcriptome Analysis.

Hanyuan Shen,R Daniel Kortschak,Thazin Nwe Aung,Jian Cui,Wei Wang,Zhipeng Qu,David L Adelson,Yuka Harata-Lee

doi:10.3389/fonc.2019.00632

Hanyuan Shen, R Daniel Kortschak + Show 6 more

Open Access

https://doi.org/10.3389/fonc.2019.00632

Copy DOI

Abstract

Herbal compatibility is the knowledge of which herbs to combine in traditional Chinese medicine (TCM) formulations. The lack of understanding of herbal compatibility is one of the key problems for the application and popularization of TCM in western society. Because of the chemical complexity of herbal medicines, it is simpler to begin to conduct compatibility research based on herbs rather than component plant secondary metabolites. We have used transcriptome analysis to explore the effects and interactions of two plant extracts (Kushen and Baituling) combined in Compound Kushen Injection (CKI). Based on shared chemical compounds and in vitro cytotoxicity comparisons, we found that both the major compounds in CKI, and the cytotoxicity effects of CKI were mainly derived from the extract of Kushen (Sophorae flavescentis). We generated and analyzed transcriptome data from MDA-MB-231 cells treated with single-herb extracts or CKI and results showed that Kushen contributed to the perturbation of the majority of cytotoxicity/cancer related pathways in CKI such as cell cycle and DNA replication. We also found that Baituling (Heterosmilax yunnanensis Gagnep) could not only enhance the cytotoxic effects of Kushen in CKI, but also activate immune-related pathways. Our analyses predicted that IL-1β gene expression was upregulated by Baituling in CKI and we confirmed that IL-1β protein expression was increased using an ELISA assay. Altogether, these findings help to explain the rationale for combining Kushen and Baituling in CKI, and show that transcriptome analysis using single herb extracts is an effective method for understanding herbal compatibility in TCM.

Highlights

At present many complex and chronic diseases rely on therapies that combine modern pharmaceuticals
By comparing transcriptome changes in MDA-MB-231 cells between Compound Kushen Injection (CKI) and single herbal extract treatments, we found that Kushen extract alone, perturbed most of the pathways through which CKI exerts its effects on cancer cells
Neither of the single injections had apoptotic effects comparable to CKI (Figure 2A) based on rates of apoptosis determined by flow cytometry with propidium iodide (PI) staining

Summary

Introduction

At present many complex and chronic diseases rely on therapies that combine modern pharmaceuticals. Recent rapid developments in analytical chemistry and molecular biology have provided methods for researchers to tackle the complex mechanisms of herbal compatibility on a number of different levels These methods focus on one or a few components within a complex mixture, in attempts to reveal how preparation/extraction for combined use can change their concentrations in products or pharmacokinetic processes in vivo [6,7,8,9]. New high-throughput technologies for measuring molecular phenotypes such as gene expression, and bioinformatic methods can provide systematic ways for refining and clarifying complex biological processes that result from hundreds or thousands of molecular interactions By applying these methods to the study of TCM, it is possible to transform the research paradigm from “main active compound that influences one target” to “multiple components that influence many network targets” [10, 11]. We apply transcriptome analysis to identify how the combination of Kushen (Sophorae flavescentis) and Baituling (Heterosmilax yunnanensis Gagnep) extracts can account for the broader and increased effects observed in Compound Kushen Injection (CKI)

Methods

Results

Conclusion