IMMUNOMODULATORY EFFECT OF DIFFERENT PROPORTIONS OF THE HERBAL MIXTURE IN TRIPHALA ON HUMAN T LYMPHOCYTES (MOLT-4)

Objectives: To investigate effect of different proportions of Terminalia bellerica Roxb (TB):Terminalia chebula Retz (TC):Phyllanthus emblica L. (PE) in triphala extract on immunological activity and to determine the possibility of applying the appropriate extract for health and immune disorders.Methods: The extracts with different proportions of TB: TC: PE (w/w/w), i.e.12:8:4 (F1), 4:12:8 (F2), 8:4:12 (F3) and 8:8:8 (F4), were prepared by decoction in water and dried under vacuum. Gallic acid, a major compound in triphala, was detected by high performance liquid chromatography (HPLC). The effect of the extracts on IFN-g and IL-10 cytokine production produced by MOLT-4 cells was determined by ELISA.Results: The results show that F1, F4 and TB extracts significantly stimulated IFN-g production but no alteration in IL-10 expression was observed. With LPS induction, F1, F2, F3 and PE extracts significantly inhibited IFN-g production, while F2, F3, TB and TC extracts inhibited IL-10 production. By determining the IFN-g/IL-10 cytokine ratio, we found that the Th1/Th2 balance after treatment with triphala extract was mainly skewed toward a Th1-like response. With LPS induction, only the F1 extract could restore the balance of immunity by shifting the Th2 response to a normal level.Conclusion: Our investigation indicates that different proportions of triphala extracts and induction conditions affect cytokine production, with a predominant Th1 response. F4, the equal proportion triphala extract, could be applied as a healthy herbal drink. F1, containing a high proportion of T. bellerica, was a promising extract as an effective therapeutic intervention against Th2 imbalance diseases such as allergy and autoimmune disease or for use with cancer vaccines.Â

IMMUNOMODULATORY ACTIVITY OF AN ACETONE EXTRACT OF TERMINALIA BELLERICA ROXB FRUIT ON THE MOUSE IMMUNE RESPONSE IN VITRO

Objectives: To investigate the immunomodulatory activity of an acetone extract of T. bellerica fruit. Methods: Mitogen induced-lymphocyte proliferation using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) technique, Th1- and Th2-related cytokine production by lymphocytes using ELISA and peritoneal macrophage function in ICR mice were assayed. Results: The results show that the extract had a mild inhibitory effect on the generation of oxidase enzyme (Phagocytic Index 0.8, 100 mg/ml) but did not influence acid phosphatase enzyme function during phagocytosis. The extract stimulated the proliferation of both T and B lymphocytes. The maximal activation (Stimulation Index 3.2, 100 mg/ml) was presented with concanavalin A induction, indicating a major effect on T lymphocyte proliferation. The extract reduced the production of IFN-γ (89%, 100 mg/ml) and IL-2 (98%, 100 mg/ml) but increased IL-10 secretion (231%, 100 mg/ml) compared to concanavalin A. Gallic acid, a pharmacological component contained in this plant, presented a similar effect as that of T. bellerica extract and may contribute to the immunomodulatory activity of T. bellerica fruits in cooperation with other phytocompounds. The decrease in the IFN-g/IL-10 ratio indicated a shift in the Th1/Th2 balance towards a Th2-type response, which might lead to a treatment for Th1-mediated inflammatory immune diseases. Conclusion: Our investigations show that the acetone extract of T. bellerica fruit possesses immunomodulatory activity, which could be used to explain its folklore applications and provide a pharmacological basis for its usefulness in immune-related disorders

INHIBITION OF LIPID ACCUMULATION IN 3T3-L1 ADIPOCYTES BY MORINDA CITRIFOLIA LINN. LEAF EXTRACTS AND COMMERCIAL HERBAL FORMULAS FOR WEIGHT CONTROL

Objective: To determine the in vitro anti-obesity effects of Morinda citrifolia leaf extract and herbal formulas used for weight loss in Thailand on lipid accumulation in 3T3-L1 adipocytes.Methods: Differentiated 3T3-L1 adipocytes were treated with 0.1, 0.5 and 1 mg/ml M. citrifolia leaf extract, three herbal formulas (JL-RU, JL-TH, CD-H) and 1, 5 and 10 mg/ml rutin, gallic acid and caffeine. Lipid accumulation determined by measuring Oil Red O staining and triglyceride content measured by a colorimetric method in adipocytes were assayed compared to the control. The effect of test samples on the viability of preadipocytes and differentiated adipocytes were investigated.Results: Differentiated adipocytes treated with 1 mg/ml M. citrifolia extract moderately inhibited fat accumulation (45.12%) and highly reduced triglyceride content (85.09%). Among the three herbal formulas, JL-TH considerably inhibited fat accumulation (109.17%, 1 mg/ml) and decreased triglyceride content (95.00%, 1 mg/ml) in adipocytes; this was higher than that for CD-H and JL-RU, respectively. The viability of preadipocytes treated with CD-H at 1 mg/ml was slightly decreased while those treated with JL-TH at 0.05-1 mg/ml showed moderately decreased viability in a dose-response manner. For differentiated adipocytes, CD-H at 0.5-1 mg/ml moderately decreased cell viability while JL-TH at 0.05-1 mg/ml caused moderate to high reduction of cell viability in a dose-response relationship.Conclusion: M. citrifolia extract and three herbal formulas had anti-obesity effects in 3T3-L1 adipocytes, as indicated by a significant reduction in lipid accumulation, triglyceride content, and cell viability. These findings suggest a potential therapeutic approach for the prevention and treatment of obesity