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Immunomodulatory Effect of Gymnema sylvestre (R.Br.) Leaf Extract: An In Vitro Study in Rat Model.

Singh VK, Dwivedi P, Chaudhary BR, Singh R - PLoS ONE (2015)

Bottom Line: EC50 value was 3.10, 3.75 and 2.68 μg/ml for NBT reduction, nitrite release and lymphoproliferation, respectively.These findings suggest the presence of active compounds, gymnemic acid, in methanolic extract of G. sylvestre leaf that stimulates both myeloid and lymphoid components of immune system, and therefore can restore the innate immune function.Through this study, the traditional knowledge of anti-diabetic property of G. sylvestre is scientifically supplemented with its immunomodulatory properties.

View Article: PubMed Central - PubMed

Affiliation: Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi, U.P., India.

ABSTRACT
Gymnema sylvestre Wild R.Br (family: Asclepidaceae) is a valuable medicinal plant used in folk medicine to treat diabetes, obesity, asthma etc. in India for antiquity. Diabetes mellitus is a syndrome characterized immunologically by lymphocyte apoptosis and reduced cell-mediated and humoral immunity. Modulation of immune responses to alleviate diseases has been of interest, and traditional herbal medicines may play an important role in this regard. In this study, we aim to evaluate the immunomodulatory potential of methanolic extract of G. sylvestre leaf using rat model. HPLC analysis of leaf extract was carried out for gymnemic acid. The method involves the initial hydrolysis of gymnemic acids, the active ingredients, to a common aglycone followed by the quantitative estimation of gymnemagenin, using gymnemagenin as reference standard. Gymnemic acid content was 2.40% (w/w) in G. sylvestre leaf extract. In vitro immunomodulatory activity of the methanolic extract of G. sylvestre leaf (1-200μg/ml) was evaluated by gauging its effects on nitroblue tetrazolium reduction and nitrite release in rat peritoneal macrophages and on mitogen (ConA, PHA and LPS) induced splenic lymphocyte proliferation. G. sylvestre leaf extract showed significant (<0.05) enhancement in NO and ROS generation in macrophages and in proliferation of lymphocytes in dose dependent manner. EC50 value was 3.10, 3.75 and 2.68 μg/ml for NBT reduction, nitrite release and lymphoproliferation, respectively. Potential effect was observed at 100 μg/ml in NO and ROS generation in macrophages and 20 μg/ml in lymphocyte proliferation. G. sylvestre leaf extract stimulates macrophage reactivity, increasing the level of activity even higher when combined with PMA or LPS. These findings suggest the presence of active compounds, gymnemic acid, in methanolic extract of G. sylvestre leaf that stimulates both myeloid and lymphoid components of immune system, and therefore can restore the innate immune function. Through this study, the traditional knowledge of anti-diabetic property of G. sylvestre is scientifically supplemented with its immunomodulatory properties.

No MeSH data available.


Related in: MedlinePlus

NBT reduction in rat peritoneal macrophages in vitro treated with Gymnema sylvestre leaf extract alone (A) and in combination with PMA 1μg/ml (B) and LPS 1μg/ml (C).GS1 –GS200 represent concentration of extract, 1–200μg/ml, respectively. Bars bearing same alphabet do not differ significantly. [Treatment effect ANOVA: F = 10.69 (8,18); p<0.05, (B) F = 7.57 (7,16); p<0.05, (C) F = 14.27 (7,16); p<0.05].
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pone.0139631.g001: NBT reduction in rat peritoneal macrophages in vitro treated with Gymnema sylvestre leaf extract alone (A) and in combination with PMA 1μg/ml (B) and LPS 1μg/ml (C).GS1 –GS200 represent concentration of extract, 1–200μg/ml, respectively. Bars bearing same alphabet do not differ significantly. [Treatment effect ANOVA: F = 10.69 (8,18); p<0.05, (B) F = 7.57 (7,16); p<0.05, (C) F = 14.27 (7,16); p<0.05].

Mentions: GS extract stimulated NBT reduction in a concentration dependent manner up to 50 μg/ml (R2 = 0.85, y = 0.74x–33.90; p<0.05), being maximal (120%) at 100 μg//ml, above which no further increased stimulation was observed (Fig 1A). The EC50 value was 3.10μg/ml. The stimulation of NBT reduction was significantly (p<0.05) higher at 50, 100, 200μg/ml extract concentration than that caused by standard immunoelicitor, PMA or LPS; while 5, 10 and 20 μg/ml, showed stimulatory effect similar to that of LPS or PMA; and 1 μg/ml extract, less than that. When macrophages incubated with extract in presence of PMA or LPS, a higher level NBT dye reduction, though insignificant, occurred at each concentration. Maximal NBT reduction was observed at 50, 100 and 200 μg/ml extract concentration in presence of PMA or LPS; while 1, 5, 10 and 20μg/ml showed as much reduction as shown by PMA (Fig 1B) or LPS alone (Fig 1C).


Immunomodulatory Effect of Gymnema sylvestre (R.Br.) Leaf Extract: An In Vitro Study in Rat Model.

Singh VK, Dwivedi P, Chaudhary BR, Singh R - PLoS ONE (2015)

NBT reduction in rat peritoneal macrophages in vitro treated with Gymnema sylvestre leaf extract alone (A) and in combination with PMA 1μg/ml (B) and LPS 1μg/ml (C).GS1 –GS200 represent concentration of extract, 1–200μg/ml, respectively. Bars bearing same alphabet do not differ significantly. [Treatment effect ANOVA: F = 10.69 (8,18); p<0.05, (B) F = 7.57 (7,16); p<0.05, (C) F = 14.27 (7,16); p<0.05].
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608767&req=5

pone.0139631.g001: NBT reduction in rat peritoneal macrophages in vitro treated with Gymnema sylvestre leaf extract alone (A) and in combination with PMA 1μg/ml (B) and LPS 1μg/ml (C).GS1 –GS200 represent concentration of extract, 1–200μg/ml, respectively. Bars bearing same alphabet do not differ significantly. [Treatment effect ANOVA: F = 10.69 (8,18); p<0.05, (B) F = 7.57 (7,16); p<0.05, (C) F = 14.27 (7,16); p<0.05].
Mentions: GS extract stimulated NBT reduction in a concentration dependent manner up to 50 μg/ml (R2 = 0.85, y = 0.74x–33.90; p<0.05), being maximal (120%) at 100 μg//ml, above which no further increased stimulation was observed (Fig 1A). The EC50 value was 3.10μg/ml. The stimulation of NBT reduction was significantly (p<0.05) higher at 50, 100, 200μg/ml extract concentration than that caused by standard immunoelicitor, PMA or LPS; while 5, 10 and 20 μg/ml, showed stimulatory effect similar to that of LPS or PMA; and 1 μg/ml extract, less than that. When macrophages incubated with extract in presence of PMA or LPS, a higher level NBT dye reduction, though insignificant, occurred at each concentration. Maximal NBT reduction was observed at 50, 100 and 200 μg/ml extract concentration in presence of PMA or LPS; while 1, 5, 10 and 20μg/ml showed as much reduction as shown by PMA (Fig 1B) or LPS alone (Fig 1C).

Bottom Line: EC50 value was 3.10, 3.75 and 2.68 μg/ml for NBT reduction, nitrite release and lymphoproliferation, respectively.These findings suggest the presence of active compounds, gymnemic acid, in methanolic extract of G. sylvestre leaf that stimulates both myeloid and lymphoid components of immune system, and therefore can restore the innate immune function.Through this study, the traditional knowledge of anti-diabetic property of G. sylvestre is scientifically supplemented with its immunomodulatory properties.

View Article: PubMed Central - PubMed

Affiliation: Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi, U.P., India.

ABSTRACT
Gymnema sylvestre Wild R.Br (family: Asclepidaceae) is a valuable medicinal plant used in folk medicine to treat diabetes, obesity, asthma etc. in India for antiquity. Diabetes mellitus is a syndrome characterized immunologically by lymphocyte apoptosis and reduced cell-mediated and humoral immunity. Modulation of immune responses to alleviate diseases has been of interest, and traditional herbal medicines may play an important role in this regard. In this study, we aim to evaluate the immunomodulatory potential of methanolic extract of G. sylvestre leaf using rat model. HPLC analysis of leaf extract was carried out for gymnemic acid. The method involves the initial hydrolysis of gymnemic acids, the active ingredients, to a common aglycone followed by the quantitative estimation of gymnemagenin, using gymnemagenin as reference standard. Gymnemic acid content was 2.40% (w/w) in G. sylvestre leaf extract. In vitro immunomodulatory activity of the methanolic extract of G. sylvestre leaf (1-200μg/ml) was evaluated by gauging its effects on nitroblue tetrazolium reduction and nitrite release in rat peritoneal macrophages and on mitogen (ConA, PHA and LPS) induced splenic lymphocyte proliferation. G. sylvestre leaf extract showed significant (<0.05) enhancement in NO and ROS generation in macrophages and in proliferation of lymphocytes in dose dependent manner. EC50 value was 3.10, 3.75 and 2.68 μg/ml for NBT reduction, nitrite release and lymphoproliferation, respectively. Potential effect was observed at 100 μg/ml in NO and ROS generation in macrophages and 20 μg/ml in lymphocyte proliferation. G. sylvestre leaf extract stimulates macrophage reactivity, increasing the level of activity even higher when combined with PMA or LPS. These findings suggest the presence of active compounds, gymnemic acid, in methanolic extract of G. sylvestre leaf that stimulates both myeloid and lymphoid components of immune system, and therefore can restore the innate immune function. Through this study, the traditional knowledge of anti-diabetic property of G. sylvestre is scientifically supplemented with its immunomodulatory properties.

No MeSH data available.


Related in: MedlinePlus