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Screening of Crude Drugs Used in Japanese Kampo Formulas for Autophagy-Mediated Cell Survival of the Human Hepatocellular Carcinoma Cell Line

1
Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Nagasaki International University, Nagasaki 859-3298, Japan
2
Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagasaki International University, Nagasaki 859-3298, Japan
*
Author to whom correspondence should be addressed.
Medicines 2019, 6(2), 63; https://doi.org/10.3390/medicines6020063
Received: 5 May 2019 / Revised: 27 May 2019 / Accepted: 28 May 2019 / Published: 3 June 2019
(This article belongs to the Special Issue Plant-Derived Medicines and Biological Activity Evaluation)
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Abstract

Background: Autophagy is a catabolic process through which dysfunctional proteins and organelles are degraded, and that is associated with the proliferation of cancer cells. The aim of this study was to screen approximately 130 kinds of crude drugs used in Japanese Kampo formulas to identify crude drugs that would regulate the proliferation through autophagy of human hepatocellular carcinoma HepG2 cells. Methods: Extracts of each crude drug were prepared using methanol. Protein levels were determined using Western blotting. Cell viability was measured by MTT assay. Results: Among the 130 crude extracts, 24 of them increased LC3-II expression. Among these, Goboshi (burdock fruit), Soboku (sappan wood), Mokko (saussurea root), Rengyo (forsythia fruit), and Hikai (dioscorea) notably suppressed the proliferation of HepG2 cells and increased p62 expression levels, which suggested that these five extracts downregulate the autophagic activity resulting in the accumulation of p62. On the other hand, Hishinomi (water chestnut), Biwayo (loquat leaf), and Binroji (areca) induced cell growth and decreased or were uninvolved with p62 expression levels, which implied that these three extracts might induce autophagy modulators for cell growth. Conclusions: The results suggest that the compounds contained in the crude drugs selected for this study could control cell viability by regulating autophagic activity in HepG2 cells. The isolation and identification of the active compounds in these drugs might lead to the development of agents for autophagy research and cancer chemoprevention. View Full-Text
Keywords: autophagic activity; screening; crude drugs; Kampo medicines; cell viability; LC3-II; p62; HepG2 autophagic activity; screening; crude drugs; Kampo medicines; cell viability; LC3-II; p62; HepG2
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Okubo, S.; Komori, H.; Kuwahara, A.; Ohta, T.; Shoyama, Y.; Uto, T. Screening of Crude Drugs Used in Japanese Kampo Formulas for Autophagy-Mediated Cell Survival of the Human Hepatocellular Carcinoma Cell Line. Medicines 2019, 6, 63.

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