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Cytotoxic effects of oosporein isolated from endophytic fungus Cochliobolus kusanoi.

Ramesha A, Venkataramana M, Nirmaladevi D, Gupta VK, Chandranayaka S, Srinivas C - Front Microbiol (2015)

Bottom Line: After 24 h of exposure histopathological observations were made to know the effects of oosporein on target organs.Further, oosporein treatment to studied cell lines indicated significant suppression of oxidative stress related gene (Superoxide dismutase1 and Catalase ) expression, and increased levels of mRNA expression in apoptosis or oxidative stress inducing genes HSP70, Caspase3, Caspase6, and Caspase9 as measured by quantitative real time-PCR assay.The high previlance of these fungi in temperate climates further warrants the need of safe food grain storage and processing practices to control the toxic effects of oosporein to humans and live stock.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Biotechnology, Bangalore University Bangalore, India.

ABSTRACT
In the present study, oosporein, a fungal toxic secondary metabolite known to be a toxic agent causing chronic disorders in animals, was isolated from fungus Cochliobolus kusanoi of Nerium oleander L. Toxic effects of oosporein and the possible mechanisms of cytotoxicity as well as the role of oxidative stress in cytotoxicity to Madin-Darby canine kidney kidney cells and RAW 264.7 splene cells were evaluated in vitro. Also to know the possible in vivo toxic effects of oosporein on kidney and spleen, Balb/C mouse were treated with different concentrations of oosporein ranging from 20 to 200 μM). After 24 h of exposure histopathological observations were made to know the effects of oosporein on target organs. Oosporein induced elevated levels of reactive oxygen species (ROS) generation and high levels of malondialdehyde, loss of mitochondrial membrane potential, induced glutathione hydroxylase (GSH) production was observed in a dose depended manner. Effects oosporein on chromosomal DNA damage was assessed by Comet assay, and increase in DNA damage were observed in both the studied cell lines by increasing the oosporein concentration. Further, oosporein treatment to studied cell lines indicated significant suppression of oxidative stress related gene (Superoxide dismutase1 and Catalase ) expression, and increased levels of mRNA expression in apoptosis or oxidative stress inducing genes HSP70, Caspase3, Caspase6, and Caspase9 as measured by quantitative real time-PCR assay. Histopathological examination of oosporein treated mouse kidney and splenocytes further revealed that, oosporein treated target mouse tissues were significantly damaged with that of untreated sam control mice and these effects were in directly proportional to the the toxin dose. Results of the present study reveals that, ROS is the principle event prompting increased oosporein toxicity in studied in vivio and in vitro animal models. The high previlance of these fungi in temperate climates further warrants the need of safe food grain storage and processing practices to control the toxic effects of oosporein to humans and live stock.

No MeSH data available.


Related in: MedlinePlus

(A) Dose dependent cytotoxic effect of oosporein on Madin-Darby canine kidney (MDCK) and RAW 264.7 cell proliferation; (B) Dose dependent effect of oosporein on plasma membrane damage in MDCK and RAW 264.7 celllines. Values represent mean ± SD of six parallel experiments. In the figure each series mean values followed by the different letter are significantly different according to DMRT at p < 0.05.
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Figure 1: (A) Dose dependent cytotoxic effect of oosporein on Madin-Darby canine kidney (MDCK) and RAW 264.7 cell proliferation; (B) Dose dependent effect of oosporein on plasma membrane damage in MDCK and RAW 264.7 celllines. Values represent mean ± SD of six parallel experiments. In the figure each series mean values followed by the different letter are significantly different according to DMRT at p < 0.05.

Mentions: The viability of MDCK and RAW 264.7 cells was measured after exposure to oosporein for 3 and 24 h by the method for the determination of the cell numbers by MTT assay. The cell feasibility assessed at 3 and 24 h after the initial introduction of the compound was reduced with increasing concentration. The viability of the cells in control and treated were significantly different in both the exposures. Besides, an increased cell death was observed in oosporein-treated cells as compared to control cells in both the cell lines. The 50% inhibitory concentration (IC50) for 24 h was found to be 86 μM for MDCK cells and 78 μM for RAW264.7 cells. Whereas for 3 h exposure time the IC50 was found to be at over 150 μM. The higher the oosporein concentration, increase in the cell death rate (Figure 1A).


Cytotoxic effects of oosporein isolated from endophytic fungus Cochliobolus kusanoi.

Ramesha A, Venkataramana M, Nirmaladevi D, Gupta VK, Chandranayaka S, Srinivas C - Front Microbiol (2015)

(A) Dose dependent cytotoxic effect of oosporein on Madin-Darby canine kidney (MDCK) and RAW 264.7 cell proliferation; (B) Dose dependent effect of oosporein on plasma membrane damage in MDCK and RAW 264.7 celllines. Values represent mean ± SD of six parallel experiments. In the figure each series mean values followed by the different letter are significantly different according to DMRT at p < 0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556033&req=5

Figure 1: (A) Dose dependent cytotoxic effect of oosporein on Madin-Darby canine kidney (MDCK) and RAW 264.7 cell proliferation; (B) Dose dependent effect of oosporein on plasma membrane damage in MDCK and RAW 264.7 celllines. Values represent mean ± SD of six parallel experiments. In the figure each series mean values followed by the different letter are significantly different according to DMRT at p < 0.05.
Mentions: The viability of MDCK and RAW 264.7 cells was measured after exposure to oosporein for 3 and 24 h by the method for the determination of the cell numbers by MTT assay. The cell feasibility assessed at 3 and 24 h after the initial introduction of the compound was reduced with increasing concentration. The viability of the cells in control and treated were significantly different in both the exposures. Besides, an increased cell death was observed in oosporein-treated cells as compared to control cells in both the cell lines. The 50% inhibitory concentration (IC50) for 24 h was found to be 86 μM for MDCK cells and 78 μM for RAW264.7 cells. Whereas for 3 h exposure time the IC50 was found to be at over 150 μM. The higher the oosporein concentration, increase in the cell death rate (Figure 1A).

Bottom Line: After 24 h of exposure histopathological observations were made to know the effects of oosporein on target organs.Further, oosporein treatment to studied cell lines indicated significant suppression of oxidative stress related gene (Superoxide dismutase1 and Catalase ) expression, and increased levels of mRNA expression in apoptosis or oxidative stress inducing genes HSP70, Caspase3, Caspase6, and Caspase9 as measured by quantitative real time-PCR assay.The high previlance of these fungi in temperate climates further warrants the need of safe food grain storage and processing practices to control the toxic effects of oosporein to humans and live stock.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Biotechnology, Bangalore University Bangalore, India.

ABSTRACT
In the present study, oosporein, a fungal toxic secondary metabolite known to be a toxic agent causing chronic disorders in animals, was isolated from fungus Cochliobolus kusanoi of Nerium oleander L. Toxic effects of oosporein and the possible mechanisms of cytotoxicity as well as the role of oxidative stress in cytotoxicity to Madin-Darby canine kidney kidney cells and RAW 264.7 splene cells were evaluated in vitro. Also to know the possible in vivo toxic effects of oosporein on kidney and spleen, Balb/C mouse were treated with different concentrations of oosporein ranging from 20 to 200 μM). After 24 h of exposure histopathological observations were made to know the effects of oosporein on target organs. Oosporein induced elevated levels of reactive oxygen species (ROS) generation and high levels of malondialdehyde, loss of mitochondrial membrane potential, induced glutathione hydroxylase (GSH) production was observed in a dose depended manner. Effects oosporein on chromosomal DNA damage was assessed by Comet assay, and increase in DNA damage were observed in both the studied cell lines by increasing the oosporein concentration. Further, oosporein treatment to studied cell lines indicated significant suppression of oxidative stress related gene (Superoxide dismutase1 and Catalase ) expression, and increased levels of mRNA expression in apoptosis or oxidative stress inducing genes HSP70, Caspase3, Caspase6, and Caspase9 as measured by quantitative real time-PCR assay. Histopathological examination of oosporein treated mouse kidney and splenocytes further revealed that, oosporein treated target mouse tissues were significantly damaged with that of untreated sam control mice and these effects were in directly proportional to the the toxin dose. Results of the present study reveals that, ROS is the principle event prompting increased oosporein toxicity in studied in vivio and in vitro animal models. The high previlance of these fungi in temperate climates further warrants the need of safe food grain storage and processing practices to control the toxic effects of oosporein to humans and live stock.

No MeSH data available.


Related in: MedlinePlus