Page 139 - journal-7-2-S-Full

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PP-21

Effect of Vernonia cinerea (L.) Less methanolic extract on in vitro

immunomodulatory activity

1,2 3 2 2,4
Aurasorn Saraphanchotiwitthaya , Thanasak Teaktong , Janya Ruenkesorn , Pattana Sripalakit
1
Department of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences,
2
Pharmaceutical Biotechnology Research Unit, Faculty of Pharmaceutical Sciences,
3
Department of Pharmacy Practice, Faculty of Pharmaceutical Sciences,
4
Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences, Naresuan Univer-
sity, Phitsanulok 65000, Thailand

Rational: Vernonia cinerea (Asteraceae) is an herbaceous plant commonly found in Thailand. Different
parts of V. cinerea have been used in folklore applications and have been reported to possess medicinal
properties relating to the immunological responses such as inflammation, infection, liver disease and asthma.
Therefore, traditional applications of V. cinerea on anti-inflammatory and immunomodulatory activity were
investigated.
Objective: To investigate the effects of V. cinerea methanolic extract on the mouse immune system in
vitro.
Methodology: The dried aerial parts of V. cinerea were extracted by maceration in methanol and a yield
of 15% (w / w of dried material) was obtained. V. cinerea e extracts (0.01, 0.1, 1, 10 and 100 μg / ml) were
examined on the production of four Th1- and Th2-related cytokines (IFN- γIL-2, IL-4, IL-10) using ELISA, and the
proliferative response of mouse splenic lymphocytes using MTT [3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide] assays. We also evaluated phagocytic activity of peritoneal mouse macrophages, human THP-1 cells
using nitroblue tetrazolium (NBT) dye reduction and cellular lysosomal enzyme activity assays.
Results: V. cinerea extract inhibited Th1-related cytokine release of IFN-γ and IL-2, and selectively inhib-
ited Th2-related cytokine secretion; IL-10 production was increased, and IL-4 production was decreased. MTT
assays were used to examine mouse splenocyte proliferation in response to phytohemagglutinin (PHA) and
concanavalin A (con A) as T cell mitogens, and lipopolysaccharide (LPS) and pokeweed mitogen (PWM) as B
cell mitogens. The extract exerted a dual effect on T cell proliferation through the same mechanism as con A,
but did not show a significant difference in the presence of PHA. Moreover, the extract inhibited B cell
proliferation through a T cell independent pathway shared with LPS, but stimulated B cell proliferation through
a T cell dependent mechanism shared with PWM. Our results suggested that V. cinerea-induced cytokine
production might be correlated with lymphocyte proliferation, caused by the activation of T cells into Th2 cells
accompanied by the suppression of Th1 cell proliferation. This hypothesis was supported by a significant
decrease in the IFN-γ / IL-10 ratio. Moreover, the extract decreased superoxide production by mouse macroph-
ages, but increased production by human THP-1 cells without significant effects on acid phosphatase activity.
These results suggest that V. cinerea may induce species-specific phagocytic responses that may partially
contribute to the plantûs anti-inflammatory activity.
Conclusion: Our findings confirmed the traditional applications of V. cinerea on anti-inflammatory and
immunomodulatory activity. Our results suggested a potential therapeutic application of this plant in the
treatment of diseases associated with Th1-related cytokine production and inflammation. In vivo immunological

assays and the underlying mechanisms of action should be further studied.