Gallic acid and MiADMSA reversed arsenic induced oxidative/nitrosative damage in rat red blood cells

Abstract

Arsenic (As) is naturally occurring toxic metalloid which is considered as a serious environmental and health concern. Red blood cells are the prime target for any toxicants as their population is higher in systemic circulation. High prevalence of anaemia too has been reported from arsenic contaminated area, suggesting possible linkage between arsenic and the damaging effects on RBCs. The exact mechanism for these effects is still not clear, however, oxidative/nitrosative stress might be one of the causative factors to play a key role. The present study was planned to evaluate the protective effects of a metal chelator, MiADMSA either alone or in combination with a natural antioxidant (gallic acid) for the reversal of arsenic induced oxidative damage in red blood cells. We collected rat RBCs and cultured them in appropriate medium. They were incubated with MiADMSA and gallic acid and then treated with sodium arsenite at 37 °C. Hemolysates were prepared and assayed for various biochemical parameters such as oxidative/nitrosative variables, osmotic fragility, acetylcholinesterase activity, and cellular metal accumulation. We found there was reversibility of oxidative/nitrosative stress variables, elevated cellular antioxidant power, and decreased osmotic fragility of red blood cells both in MiADMSA alone as well as in combination with gallic acid treated group compared with arsenic treated group. In conclusion, MiADMSA efficiently participated in the reversal of arsenic induced oxidative/nitrosative damage in red blood cells where as Gallic acid improved its reversal when given in combination with MiADMSA.

Keywords: Antioxidant; Behavioral neuroscience; Developmental biology; Flavonoid; Gallic acid; MiADMSA; Nervous system; Neurotoxicology; Oxidative stress; Oxidative/nitrosative stress; Pathophysiology; Photochemical; Reactive oxygen species; Red blood cells; Sodium arsenite; Toxicology.

Figure 1

Concentration of arsenic in whole blood at different time interval after single dose oral administration in male Wistar rats.

Figure 2

Concentration of arsenic in RBCs of various groups. All the values are expressed as mean ± SEM (n = 3), ****P < 0.0001 vs control, ^#P < 0.05, ^##P < 0.01 vs arsenic and ^$$$$P < 0.0001 vs As + MiADMSA. [GA = gallic acid, MiADMSA = monoisoamyl DMSA, As = arsenic].

Figure 3

Effect of arsenic exposure on osmotic fragility. All the values are expressed as mean ± SEM (n = 3), ****P < 0.0001 vs. Control and ^####P < 0.0001 vs. Arsenic and ^$$$$P < 0.0001 vs. Arsenic + MiADMSA.

Figure 4

A) Effect of arsenic exposure and protective therapies on ROS level B) Effect of arsenic exposure and protective therapies on antioxidant power. All the values are expressed as mean ± SEM (n = 3), **P < 0.01, ****P < 0.0001 vs. Control and #P < 0.05, ^##P < 0.01, ^####P < 0.0001 vs. Arsenic ^$$P < 0.01 vs. Arsenic + MiADMSA.

Figure 5

A) Effect of arsenic exposure and protective therapies on MDA level B) Effect of arsenic exposure and protective therapies on reduced GSH level. C) Effect of arsenic exposure and protective therapies on catalase activity. D) Effect of arsenic exposure and protective therapies on SOD activity. E) Effect of arsenic exposure and protective therapies on nitrite level. All the values are expressed as mean ± SEM (n = 3), ****P < 0.0001 vs. Control and #P < 0.05, ##P < 0.01, ####P < 0.0001 vs. Arsenic and $$$P < 0.0001 vs. Arsenic + MiADMSA.

Figure 6

Effect of arsenic exposure and protective therapies on AChE activity. All the values are expressed as mean ± SEM (n = 3), ***P < 0.001, vs. Control and ####P < 0.0001 vs. Arsenic and $$$$P < 0.0001 vs. Arsenic + MiADMSA.

Figure 7

Inverted microscopic images of RBCs. (A) Untreated control cells. RBCs treated with (B) MiADMSA alone (C) Gallic acid alone and (D) SA alone (E) combination of SA and MiADMSA (F) combination of Gallic acid and MiADMSA (G) combination of SA, MiADMSA and Gallic acid. Morphological view of all group have taken on 100X magnification. Contrast and brightness was adjusted by Microsoft picture manager 2007.