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Session 3: Preclinical studies of herbal and traditional medicine

In vitro Anti-inflammatory, Mutagenic and Antimutagenic Acti-

vity of Cha-Tu-Ka-La-Thad Remedy and Its Ingredients

Pravaree Phuneerub*, Wacharee Limpanasithikul**, Chanida palanuvej***,
,
Nijsiri Rueangrungsri*******
*School of Health Sciences, Mae Fah Luang University, Chiang Rai 57100, Thailand
**Department of Pharmacology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thai-
land
***College of Public Health Sciences, Chulalongkorn University, Bangkok 10330, Thailand
****Faculty of Pharmacy, Rangsit University, Pathumthani 12000, Thailand

Background and rationale: Cha-Tu-Ka-La-Thad (CKT) remedy is composed of the roots
of Plumbago indica L. (Plumbaginacea), Acorus calamus L. (Araceae), Clerodendrum paniculatum
L. (Verbernaceae) and Dolichandrone serrulata (Wall. ex DC.) Seem. (Bignoniaceae), each in
an equal part by weights. CKT has been used as an antipyretic and anti-inflammatory drug in
traditional Thai medicine. However, there is no evidence to support anti-inflammatory activity
of this traditional remedy yet.
Objectives: This present study investigated the in vitro anti-inflammatory, mutagenic
and antimutagenic activities of the roots of four plant species and CKT remedy.
Methods: Murine macrophage J774A.1 cells were treated with the CKT remedy and its
ingredient extracts at the concentrations of 0, 6.25, 12.5, 25, 50, and 100 μg/ml in 96-well
plates. The cells were stimulated by lipopolysaccharide (LPS) to evaluate the production of
nitric oxide (NO), tumor necrosis factor-α (TNF-α) and prostaglandin E (PGE ) in the anti-
2 2
inflammatory test while the concentration of 25, 50, 100 and 200 mg/ml of CKT remedy and its
ingredient extracts were investigated the mutagenic and antimutagenic potential by the Ames
test in Salmonella typhimurium strain TA98 and TA100.
Results: The outcome of this study displayed that all root extracts and CKT remedy
significantly inhibited LPS-induced nitric oxide (NO), tumor necrosis factor-α (TNF-α) and
prostaglandin E (PGE ) production in the anti-inflammatory test. Additionally, only the
2 2
extracts of A. calamus exhibited direct mutagenicity on both strains and the fractionated
water of P. indica based on without and with nitrite treatment exhibited direct mutagenicity
on TA98 strain. Moreover, all extracts inhibited the mutagenicity of reaction of nitrite treated
1-aminopyrene on both strains in the Ames test.
Conclusion: Our findings suggest that CKT remedy and its ingredients can be further
developed as promising anti-inflammatory and antimutagenic agent.
Key words: Cha-Tu-Ka-La-Thad remedy, antimutagenic, murine macrophage anti-in-
flammatory activity, bacterial reverse mutation assay