ANTIOXIDANT STATUS of ERYTHROCYTES

Slide 1: 

A STUDY ON THE ANTIOXIDANT STATUS OF ERYTHROCYTES IN END STAGE RENAL DISEASED PATIENTS AND THE ANTIHEAMOLYTIC ACTIVITY OF Boerhaavia diffusa. PRESENTED BY VIJAYACHANDRIKA.V AND SATHYAPRIYA.K Department of Biochemistry, Bharathi women’s college, Chennai-108

Slide 2: 

OBJECTIVES OF THE STUDY The most important complications in ESRD patients are cardiovascular disease and aneamia. The increased degree of oxidative stress and the possible role of free radical in damaging RBC of ESRD patients have been well documented. The study was carried out to estimate the level of heamoglobin. The glutathione plays a vital role in free radical scavenging mechanism. The activity of antioxidant enzymes are included in the study The important aim of the present study is to analyse the antioxidant status of RBC and the role of B.diffusa in maintaining the integrity of RBC in an invitro condition.

Slide 3: 

INTRODUCTION Low antioxidant level is a major risk factor for CRF patients. Chronic renal failure induces aneamia by shortening the life span of erythrocytes due to increase in oxidative stress. Reactive oxygen species such as super oxide radical, hydrogen peroxide, hydroxyl radical, etc. contributes to inflammation and injury in CRF patients. Boerhaavia diffusa decreases the number of free radicals due to its antioxidant effect and the membrane stabilizing activity. B.diffusa reduces free radical damage by restoring glutathione level. Due to its anti-inflammatory activity it reduces inflammation and injury in CRF patients.

Slide 4: 

Boerhaavia diffusa

Slide 5: 

MATERIALS AND METHODS The level of lipid peroxides, reduced glutathione, glutathione peroxidase and glutathione-s-transferase were measured in the erythrocytes isolated from 10 patients (Group II) spectrophotometrically and compared with normal subjects (Group I). The osmotic fragility test was performed by incubating 0.5ml of packed cell isolated from healthy individual and polycystic ESRD patients blood with and without 0.5ml aqueous extract of B.diffusa and incubated for 15 minutes. The erythrocyte heamolysis induced by osmotic fragility was studied to monitor the membrane damage in invitro condition with and without the aqueous extract .

Slide 6: 

RESULT Figure I show the level of lipid peroxides in erythrocytes of Group I and Group II. When compared to healthy volunteers (Group I) the level of lipid peroxides significantly increased (p< 0.001) in ESRD patients (Group I and Group II). Figure II, III, IV shows the activity of glutathione-s-transferase, glutathione peroxidase and reduced glutathione in the erythrocytes of Group I and II. When compared to Group I the level of glutathione-s- transferase, glutathione peroxidase, and reduced glutathione significantly decreased (p<0.01) in ESRD patients (Group II). Graph I represents the percentage of heamolysis of control blood, ESRD patients blood with and without the aqueous extract of B.diffusa. It also represents the percentage heamolysis of control blood with the presence of aqueous extract of B.diffusa which serves as positive control.

Slide 7: 

FIGURE I Y-axis the level of lipid peroxide expressed in nmoles of TBA formed/mg protein

Slide 8: 

FIGURE II Y-axis represents the activity of Glutathione-S-transferase is expressed in nmoles of CDNB conjugated/min/mg Hb.

Slide 9: 

FIGURE III Y- Axis represents the activity of Glutathione peroxidase in nmoles of glutathione oxidized/min/mg of Hb.

Slide 10: 

FIGURE IV Y-Axis represents the level of Reduced Glutathione is expressed in ?g/mg Hb.

Slide 11: 

Graph I The percentage of heamolysis of control blood, polycystic kidney diseased patient’s blood with and without the aqueous extract of Boerhaavia diffusa.

Slide 12: 

DISCUSSION Red blood survival in patients undergoing heamolysis is reduced to one half to two- third of normal. Human RBC is a cell that might be altered as a consequence of free radical damage. Increased oxidative stress is considered to be one of the major risk factor in CRF patients undergoing heamodialysis . The present study shows the concomitant decrease in the activity of all the antioxidants in the erythrocytes of renal failure patients and increase in the level of lipid peroxides was observed. This might be due to increased oxidative stress. The present investigation also shows that the capability of the RBC to resist heamolysis was found to be increased on treatment of the patients blood with the aqueous extract of B.diffusa when compared to control and patients sample alone.

Slide 13: 

CONCLUSION The study shows an overall reduction in antioxidant status of the RBC in renal failure patients as compared to the normal group. B.diffusa which was known to play an excellent role in eradicating pro-oxidants and free radicals in addition was also proved to posses antiheamolytic role which might have greater implication in the future.