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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

Proliferation of splenic lymphocytes in vitro treated with Gymnema sylvestre leaf extract alone (A) and in combination with PHA 10 μg/ ml (B), ConA 5 μg/ ml (C) and LPS 10 μg/ml (D).GS1 –GS200 represent concentration of extract, 1–200μg/ml, respectively. Bars bearing same alphabet do not differ significantly. [Treatment effect ANOVA: (A) F = 10.58 (9, 20); p<0.05, (B) F = 27.28(7, 16); p<0.05, (C) F = 2.66 (7, 16); p<0.05; (D) F = 5.26 (6, 14); p<0.05].
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pone.0139631.g003: Proliferation of splenic lymphocytes in vitro treated with Gymnema sylvestre leaf extract alone (A) and in combination with PHA 10 μg/ ml (B), ConA 5 μg/ ml (C) and LPS 10 μg/ml (D).GS1 –GS200 represent concentration of extract, 1–200μg/ml, respectively. Bars bearing same alphabet do not differ significantly. [Treatment effect ANOVA: (A) F = 10.58 (9, 20); p<0.05, (B) F = 27.28(7, 16); p<0.05, (C) F = 2.66 (7, 16); p<0.05; (D) F = 5.26 (6, 14); p<0.05].

Mentions: Effect of GS extract on splenic lymphocyte proliferation is presented in Fig 3. The proliferative response was stimulated by 26, 71, 141, 184, 193, 187 and 141% in cultures treated with 1, 5, 10, 20, 50, 100 and 200 μg/ml GS extract, respectively, as compared to mitogen-stimulated proliferative responses (PHA, 53%; ConA, 93% and LPS, 96%).The stimulatory effect was concentration dependent up to 20 μg/ml (R2 = 0.93; y = 0.11x–2.82; p<0.05: and EC50 value extrapolated from the linear equation was 2.68μg/ml); no further enhanced stimulation was observed at 50 and 100 μg/ml concentrations, rather decreased at 200 μg/ml (Fig 3A). When GS extract was combined with PHA in cultures, 50 to 200 μg/ml concentrations showed higher level response than PHA alone, being maximal at 100μg/ml; while the lower concentrations, similar level response (Fig 3B). Proliferative response was significant (p<0.05) and maximal in lymphocyte culture treated with 50 μg/ml GS extract in presence of another T-cell mitogens, ConA, but decreased when treated with higher concentrations (Fig 3C). When the splenic lymphocyte cultures treated with GS extract along with B-cell mitogens (LPS), 20–100 μg/ml concentrations enhanced the proliferative response significantly (p<0.05) and maximally of identical level, as compared to LPS-stimulated one (Fig 3D).


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)

Proliferation of splenic lymphocytes in vitro treated with Gymnema sylvestre leaf extract alone (A) and in combination with PHA 10 μg/ ml (B), ConA 5 μg/ ml (C) and LPS 10 μg/ml (D).GS1 –GS200 represent concentration of extract, 1–200μg/ml, respectively. Bars bearing same alphabet do not differ significantly. [Treatment effect ANOVA: (A) F = 10.58 (9, 20); p<0.05, (B) F = 27.28(7, 16); p<0.05, (C) F = 2.66 (7, 16); p<0.05; (D) F = 5.26 (6, 14); p<0.05].
© Copyright Policy
Related In: Results  -  Collection

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

pone.0139631.g003: Proliferation of splenic lymphocytes in vitro treated with Gymnema sylvestre leaf extract alone (A) and in combination with PHA 10 μg/ ml (B), ConA 5 μg/ ml (C) and LPS 10 μg/ml (D).GS1 –GS200 represent concentration of extract, 1–200μg/ml, respectively. Bars bearing same alphabet do not differ significantly. [Treatment effect ANOVA: (A) F = 10.58 (9, 20); p<0.05, (B) F = 27.28(7, 16); p<0.05, (C) F = 2.66 (7, 16); p<0.05; (D) F = 5.26 (6, 14); p<0.05].
Mentions: Effect of GS extract on splenic lymphocyte proliferation is presented in Fig 3. The proliferative response was stimulated by 26, 71, 141, 184, 193, 187 and 141% in cultures treated with 1, 5, 10, 20, 50, 100 and 200 μg/ml GS extract, respectively, as compared to mitogen-stimulated proliferative responses (PHA, 53%; ConA, 93% and LPS, 96%).The stimulatory effect was concentration dependent up to 20 μg/ml (R2 = 0.93; y = 0.11x–2.82; p<0.05: and EC50 value extrapolated from the linear equation was 2.68μg/ml); no further enhanced stimulation was observed at 50 and 100 μg/ml concentrations, rather decreased at 200 μg/ml (Fig 3A). When GS extract was combined with PHA in cultures, 50 to 200 μg/ml concentrations showed higher level response than PHA alone, being maximal at 100μg/ml; while the lower concentrations, similar level response (Fig 3B). Proliferative response was significant (p<0.05) and maximal in lymphocyte culture treated with 50 μg/ml GS extract in presence of another T-cell mitogens, ConA, but decreased when treated with higher concentrations (Fig 3C). When the splenic lymphocyte cultures treated with GS extract along with B-cell mitogens (LPS), 20–100 μg/ml concentrations enhanced the proliferative response significantly (p<0.05) and maximally of identical level, as compared to LPS-stimulated one (Fig 3D).

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