Limits...
In vivo effects of traditional Ayurvedic formulations in Drosophila melanogaster model relate with therapeutic applications.

Dwivedi V, Anandan EM, Mony RS, Muraleedharan TS, Valiathan MS, Mutsuddi M, Lakhotia SC - PLoS ONE (2012)

Bottom Line: On the contrary, feeding larvae on normal food and adults on AR supplement had no effect on fecundity but a comparable regime of feeding on RS-supplemented food improved fecundity.RS feeding did not cause heavy metal toxicity.Thus, Drosophila, with its very rich genetic tools and well-worked-out developmental pathways promises to be a very good model for examining the cellular and molecular bases of the effects of different Ayurvedic formulations.

View Article: PubMed Central - PubMed

Affiliation: Cytogenetics Laboratory, Department of Zoology, Banaras Hindu University, Varanasi, India.

ABSTRACT

Background: Ayurveda represents the traditional medicine system of India. Since mechanistic details of therapy in terms of current biology are not available in Ayurvedic literature, modern scientific studies are necessary to understand its major concepts and procedures. It is necessary to examine effects of the whole Ayurvedic formulations rather than their "active" components as is done in most current studies.

Methods: We tested two different categories of formulations, a Rasayana (Amalaki Rasayana or AR, an herbal derivative) and a Bhasma (Rasa-Sindoor or RS, an organo-metallic derivative of mercury), for effects on longevity, development, fecundity, stress-tolerance, and heterogeneous nuclear ribonucleoprotein (hnRNP) levels of Drosophila melanogaster using at least 200 larvae or flies for each assay.

Results: A 0.5% (weight/volume) supplement of AR or RS affected life-history and other physiological traits in distinct ways. While the size of salivary glands, hnRNP levels in larval tissues, and thermotolerance of larvae/adult flies improved significantly following feeding either of the two formulations, the median life span and starvation resistance improved only with AR. Feeding on AR or RS supplemented food improved fecundity differently. Feeding of larvae and adults with AR increased the fecundity while the same with RS had opposite effect. On the contrary, feeding larvae on normal food and adults on AR supplement had no effect on fecundity but a comparable regime of feeding on RS-supplemented food improved fecundity. RS feeding did not cause heavy metal toxicity.

Conclusions: The present study with two Ayurvedic formulations reveals formulation-specific effects on several parameters of the fly's life, which seem to generally agree with their recommended human usages in Ayurvedic practices. Thus, Drosophila, with its very rich genetic tools and well-worked-out developmental pathways promises to be a very good model for examining the cellular and molecular bases of the effects of different Ayurvedic formulations.

Show MeSH

Related in: MedlinePlus

Levels of Hrp40 (Squid) are increased in formulation fed Squid-GFP larvae.Confocal projection images of GFP fluorescence (green, A–C and G–I) in polytene nuclei from late third instar larval salivary glands (A–C); DAPI-stained chromatin (D–F) is seen in red. Histograms in G show mean intensity of Squid-GFP fluorescence in SG polytene nuclei (N = 100 for each sample) from differently fed larvae. Western blot in H shows relative levels of Squid-GFP protein (Squid) and β-tubulin (β-tub) in whole proteins from differently fed (CON, RS or AR) larvae challenged with GFP antibody: the values below each lane indicate the relative levels (ratio of Squid∶β-tubulin in the blot with the value for larvae reared on regular food (CON) taken as 1.0) of Squid-GFP protein. The scale bar in A represents 5 µm and applies to A–F.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3351451&req=5

pone-0037113-g006: Levels of Hrp40 (Squid) are increased in formulation fed Squid-GFP larvae.Confocal projection images of GFP fluorescence (green, A–C and G–I) in polytene nuclei from late third instar larval salivary glands (A–C); DAPI-stained chromatin (D–F) is seen in red. Histograms in G show mean intensity of Squid-GFP fluorescence in SG polytene nuclei (N = 100 for each sample) from differently fed larvae. Western blot in H shows relative levels of Squid-GFP protein (Squid) and β-tubulin (β-tub) in whole proteins from differently fed (CON, RS or AR) larvae challenged with GFP antibody: the values below each lane indicate the relative levels (ratio of Squid∶β-tubulin in the blot with the value for larvae reared on regular food (CON) taken as 1.0) of Squid-GFP protein. The scale bar in A represents 5 µm and applies to A–F.

Mentions: To determine the cellular levels of the Hrp40 (Squid) protein, we used the Squid-GFP protein-trap allele so that the sub-cellular distribution of the Squid protein can be monitored through the GFP fluorescence. Confocal microscopic examination of GFP fluorescence in SG from late 3rd instar Squid-GFP larvae reared since hatching on regular (Control) or 0.5% AR or RS supplemented food revealed significantly increased levels of Squid in the formulation-fed larvae, with several puff regions showing massive accumulation of Squid-GFP (Fig. 6A–F) in a manner comparable to that noted above for the Hrb87F protein (Fig. 5). Comparison of mean GFP-fluorescence intensity of distal polytene nuclei in SG from differently fed larvae (Fig. 6G) confirmed that the nuclear levels of Squid protein are indeed enhanced in larval tissues from the formulation fed larvae. Detection of Squid-GFP in total larval proteins by western blotting (Fig. 6H) also confirmed this protein's elevated levels in formulation-fed larvae.


In vivo effects of traditional Ayurvedic formulations in Drosophila melanogaster model relate with therapeutic applications.

Dwivedi V, Anandan EM, Mony RS, Muraleedharan TS, Valiathan MS, Mutsuddi M, Lakhotia SC - PLoS ONE (2012)

Levels of Hrp40 (Squid) are increased in formulation fed Squid-GFP larvae.Confocal projection images of GFP fluorescence (green, A–C and G–I) in polytene nuclei from late third instar larval salivary glands (A–C); DAPI-stained chromatin (D–F) is seen in red. Histograms in G show mean intensity of Squid-GFP fluorescence in SG polytene nuclei (N = 100 for each sample) from differently fed larvae. Western blot in H shows relative levels of Squid-GFP protein (Squid) and β-tubulin (β-tub) in whole proteins from differently fed (CON, RS or AR) larvae challenged with GFP antibody: the values below each lane indicate the relative levels (ratio of Squid∶β-tubulin in the blot with the value for larvae reared on regular food (CON) taken as 1.0) of Squid-GFP protein. The scale bar in A represents 5 µm and applies to A–F.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3351451&req=5

pone-0037113-g006: Levels of Hrp40 (Squid) are increased in formulation fed Squid-GFP larvae.Confocal projection images of GFP fluorescence (green, A–C and G–I) in polytene nuclei from late third instar larval salivary glands (A–C); DAPI-stained chromatin (D–F) is seen in red. Histograms in G show mean intensity of Squid-GFP fluorescence in SG polytene nuclei (N = 100 for each sample) from differently fed larvae. Western blot in H shows relative levels of Squid-GFP protein (Squid) and β-tubulin (β-tub) in whole proteins from differently fed (CON, RS or AR) larvae challenged with GFP antibody: the values below each lane indicate the relative levels (ratio of Squid∶β-tubulin in the blot with the value for larvae reared on regular food (CON) taken as 1.0) of Squid-GFP protein. The scale bar in A represents 5 µm and applies to A–F.
Mentions: To determine the cellular levels of the Hrp40 (Squid) protein, we used the Squid-GFP protein-trap allele so that the sub-cellular distribution of the Squid protein can be monitored through the GFP fluorescence. Confocal microscopic examination of GFP fluorescence in SG from late 3rd instar Squid-GFP larvae reared since hatching on regular (Control) or 0.5% AR or RS supplemented food revealed significantly increased levels of Squid in the formulation-fed larvae, with several puff regions showing massive accumulation of Squid-GFP (Fig. 6A–F) in a manner comparable to that noted above for the Hrb87F protein (Fig. 5). Comparison of mean GFP-fluorescence intensity of distal polytene nuclei in SG from differently fed larvae (Fig. 6G) confirmed that the nuclear levels of Squid protein are indeed enhanced in larval tissues from the formulation fed larvae. Detection of Squid-GFP in total larval proteins by western blotting (Fig. 6H) also confirmed this protein's elevated levels in formulation-fed larvae.

Bottom Line: On the contrary, feeding larvae on normal food and adults on AR supplement had no effect on fecundity but a comparable regime of feeding on RS-supplemented food improved fecundity.RS feeding did not cause heavy metal toxicity.Thus, Drosophila, with its very rich genetic tools and well-worked-out developmental pathways promises to be a very good model for examining the cellular and molecular bases of the effects of different Ayurvedic formulations.

View Article: PubMed Central - PubMed

Affiliation: Cytogenetics Laboratory, Department of Zoology, Banaras Hindu University, Varanasi, India.

ABSTRACT

Background: Ayurveda represents the traditional medicine system of India. Since mechanistic details of therapy in terms of current biology are not available in Ayurvedic literature, modern scientific studies are necessary to understand its major concepts and procedures. It is necessary to examine effects of the whole Ayurvedic formulations rather than their "active" components as is done in most current studies.

Methods: We tested two different categories of formulations, a Rasayana (Amalaki Rasayana or AR, an herbal derivative) and a Bhasma (Rasa-Sindoor or RS, an organo-metallic derivative of mercury), for effects on longevity, development, fecundity, stress-tolerance, and heterogeneous nuclear ribonucleoprotein (hnRNP) levels of Drosophila melanogaster using at least 200 larvae or flies for each assay.

Results: A 0.5% (weight/volume) supplement of AR or RS affected life-history and other physiological traits in distinct ways. While the size of salivary glands, hnRNP levels in larval tissues, and thermotolerance of larvae/adult flies improved significantly following feeding either of the two formulations, the median life span and starvation resistance improved only with AR. Feeding on AR or RS supplemented food improved fecundity differently. Feeding of larvae and adults with AR increased the fecundity while the same with RS had opposite effect. On the contrary, feeding larvae on normal food and adults on AR supplement had no effect on fecundity but a comparable regime of feeding on RS-supplemented food improved fecundity. RS feeding did not cause heavy metal toxicity.

Conclusions: The present study with two Ayurvedic formulations reveals formulation-specific effects on several parameters of the fly's life, which seem to generally agree with their recommended human usages in Ayurvedic practices. Thus, Drosophila, with its very rich genetic tools and well-worked-out developmental pathways promises to be a very good model for examining the cellular and molecular bases of the effects of different Ayurvedic formulations.

Show MeSH
Related in: MedlinePlus