Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 May 4;123(1):61-7.
doi: 10.1016/j.jep.2009.02.028. Epub 2009 Mar 4.

The cytotoxicity to leukemia cells and antiviral effects of Isatis indigotica extracts on pseudorabies virus

Shih-Ling Hsuan  1 , Shih-Chieh Chang, Sheng-Yang Wang, Tien-Ling Liao, Ting-Ting Jong, Maw-Sheng Chien, Wei-Cheng Lee, Shih-Shiung Chen, Jiunn-Wang Liao
Affiliations

The cytotoxicity to leukemia cells and antiviral effects of Isatis indigotica extracts on pseudorabies virus

Shih-Ling Hsuan et al. J Ethnopharmacol. .

Abstract

Aim of the study: Isatis indigotica (I. indigotica), Cruciferae, has been used in Chinese medicine for anti-leukemia and anti-severe acute respiratory syndrome (SARS). The aim of this study was to evaluate the cytotoxicity of Isatis indigotica extracts on human leukemia cell line (HL-60) and the antiviral activity on swine pseudorabies virus (PrV) in in vitro assays.

Materials and methods: Extracts and derived fractions of Isatis indigotica were prepared from root (R) and leaf (L) using methanol (M), ethyl acetate (E) and distilled water (D). The cytotoxic effect of extracts on swine peripheral blood mononuclear cells (PBMCs) and HL-60 was assessed by MTT method. The cytopathic effect (CPE) reduction, plaque reduction and inhibition assays on viral replication, and virucidal activity were further conducted to investigate the anti-PrV activity.

Results: Indirubin, one of the biological active compounds of Isatis indigotica, had the most significant cytotoxicity on HL-60 cells and inhibitory effect on PrV replication. Extracts from roots and leaves of Isatis indigotica also presented CPE inhibition either before or after infection of PrV on porcine kidney (PK-15) cells. Leaf extracts had better virucidal activity than roots, and ethyl acetate extracts exhibited the highest efficacy among extracts tested.

Conclusion: Isatis indigotica posses a valuable virucidal effect in disease control of pseudorabies virus infection in swine.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
The chemical structure of indirubin (cited from Liau et al., 2007).
Fig. 2
Fig. 2
Effects of Isatis indigotica extracts by different solvents on PrV induced cytopathic effect. PK-15 cells were infected with 50 TCID50/well PrV and various concentrations of extracts in 96-well plates for 72 h. The inhibition effect (% of control) of RD, RM and LD extracts are shown in (A) and RE, LM, LE and indirubin in (B). Data are expressed as mean ± SD of three independent experiments with eight replicates each. R: root; L: leaves; M: MeOH-extracted; E: ethyl acetate-extracted; D: distilled water-extracted. *Indicates significant difference between control and treatment groups at p < 0.05.
Fig. 3
Fig. 3
Effects of Isatis indigotica extracts by different solvents on PrV plaque reduction. PK-15 cells were firstly infected with 100 pfu/well and various concentrations of extracts in 24-well plates for 1.5 h, washed and then incubated for 48 h. The plaque inhibition effect (% of control) of different extracts of RD, RM and LD are shown in (A) and RE, LM, LE and indirubin in (B). Data are expressed as mean ± SD of three independent experiments with eight replicates each. R: root; L: leaves; M: MeOH-extracted; E: ethyl acetate-extracted; D: distilled water-extracted. *Indicates significant difference between control and treatment groups at p < 0.05.
Fig. 4
Fig. 4
Effects of Isatis indigotica extracts by different solvents on PrV replication. PK-15 cells were firstly preinfected with 100 pfu/well for 1.5 h, washed, and then incubated with various concentrations of extracts in 24-well plates for 48 h. The viral inhibition (% of control) of RD, RM and LD extracts are shown in (A) and RE, LM, LE and indirubin in (B). Data are expressed as mean ± SD of three independent experiments with eight replicates each. R: root; L: leaves; M: MeOH-extracted; E: ethyl acetate-extracted; D: distilled water-extracted. *Indicates significant difference between control and treatment groups at p < 0.05.
Fig. 5
Fig. 5
Virucidal effects of Isatis indigotica extracts by different solvents on PrV. Different concentrations of extracts and indirubin were premixed with 5 × 106 pfu PR virus 25 °C for 6 h, and then incubated with PK-15 cells in 24-well plates for 48 h. The residual infectivity of PrV (% of control) after treatment of RD, RM and LD extracts are shown in (A) and RE, LM, LE and indirubin groups in (B). Data are expressed as mean ± SD of three independent experiments with eight replicates each. R: root; L: leaves; M: MeOH-extracted; E: ethyl acetate-extracted; D: distilled water-extracted. *Indicates significant difference between control and treatment groups at p < 0.05.
See this image and copyright information in PMC

References

    1. Ahmad A., Davies J., Randall S., Skinner G.R. Antiviral properties of extract of Opuntia streptacantha. Antiviral Research. 1996;30:75–85. - PubMed
    1. Chen L., Lin T., Zhang H.X., Su Y.B. Immune response to foot-and-mouth disease DNA vaccines can be enhanced by coinjection with the Isatis indigotica extract. Intervirology. 2005;48:207–212. - PubMed
    1. Cory A.H., Owen T.C., Barltrop J.A., Cory J.G. Use an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Communication. 1991;3:207–212. - PubMed
    1. Carlucci M.J., Ciancia M., Matulewicz M.C., Cerezo A.S., Damonte E.B. Antiherpetic activity and mode of action of natural carrageenans of diverse structural types. Antiviral Research. 1999;43:93–102. - PubMed
    1. Hoessel R., Leclerc S., Endicott J.A., Nobel M.E., Lawrie A., Tunnah P., Leost M., Damiens E., Marie D., Marko D., Niederberger E., Tang W., Eisenbrand G., Meijer L. Indirubin, the active constituent of a Chinese antileukaemia medicine, inhibits cyclin-dependent kinases. Nature Cell Biology. 1999;1:60–67. - PubMed

Publication types