Year 1996
Author LG.Korkina. G.A. Ibragimova, I.B.Afanns’ev, E.P.Krasnyuk,  and Yu.I.Kundiev
Publisher Chelating Agents in Pharmacology, Toxicology and Therapeutics; Oral Chelation in Treatment of thalassemia and Other Diseases. Symposium Pilsen, Czech Republic: Plzensky Lekarsky Sbornik Supplement 71





LG.Korkina. G.A. Ibragimova, I.B.Afanns’ev, E.P.Krasnyuk,  and Yu.I.Kundiev

Russian Institute of Pediatric Hematology Mascow. RtL sin and Institute of Occupational Health Kiev, Ukraine


It is known that exposure to coball-contaning dust is associated with the development of development of lung disease characterized in its extreme by a potential interstitial  fibrosis. At the same time.   Exposure to lead causes   hematological,    gastrointestinal,   and    neurological dysfunction.  However, despite  the difference  in their toxic manifestations, both  metal seems to be  able to stimulate the overproduction  of free radicals and destroy endogenous antioxidants. Therefore. it may be suggested that damaging free radical-mediated processes can mainly or partly he responsible for the toxic effects of both cobalt and lead. Early, we have shown [1,2] that Japanese  health food  supplementation  Bio-normalizer (a  natural nontoxic  substance  possessing  antioxidant  and  chelatory properties)  can  control  free radicals mediated processes in the in Vitro and  in in vivo systems. Because of that, we have studied the possibility of application of Bio-normalizer for suppressing the toxic activities  of cobalt and lead in human and animals.

The toxic activities of cobalt and lead were studied in animal model of lung injury. Cobalt sulfate was injected intratracheally to male Wistar rats. which then were sacrificed on 1st, 3d, 7th days.  Simultancously, cobalt  injections were  given  to  rats, which  were  fed  with  Bio-normalizer  (BN).  In  all  cases  animal  survival,  cobalt-induced   lung  edema,   and   the parameters  of  rats  free   radical  status  (GSH content,  erythrocyte  and  leukocyte   SOD activities,  and  oxygen   radical  release by  leukocytes and   alveolar  macrophages) were determined.

As it should be expected,  in all cases the administration  of cobalt sulfate to rats resulted in acute  toxic effects. Intracheal instillation of cobalt sulfate decreased  the survival of  rats by more than 50%. Feeding  rats with BN after  cobalt administration  sharply increased  the survival  rate  in  the  last  3  days. In contrast,   BN  administration before  Co  instillation suppressed  its toxic effect  in  the first 3 days. Cobalt  administration to rats  induced  lung edema  that achieved  a maximum  already on 3th  day; BN administration  diminished  it by 15-20% to the end of the experiment.

In contrast to previous findings obtained  with cobalt  instillation to hamstern [3], we found  that cobalt sharply increased  GSH  level in the rat lung. BN administration enhanced sinergistically  GSH  level  in the  cobalt-treated  animals.  SOD   activity in erythrocytes   of cobalt-treated  rat., decreased  by 1.5 – 2  times in comparison  with control  animals,  but BN admistration temporary  restored  SOD  activity on the 3d  day. SOD  activity in leukocytes practically was not affected by Co instillation. Cobalt  administration  significantly enhanced oxygen radical release by leukocytes (measured  by lucigenin-amplified  chemiluminescence, CL)  and decreased CL produced  by alveolar macrophages. BN suppressed  both inhibitory and stimulatory effects of cobalt (Table  1). This finding possibly shows  the most important beneficial effect of this nontoxic  natural  substance because  BN  turns  out  to  be  able  to suppress  the inflammation stimulated by leukocytes and increase  the phagocytic  activity of macrophages.


Table I

Lucigenin-amplified CL by leukocytes and alveolar macrophages from Co-trented rats

CL intensity in leukocytes (mV) CL   intensity   in   alveolar macrophnge  (mV)
Control rats 44±15 146±35
Co-treated rats 75±20 82±16
Co-treated rats +BN 56±10 134±25


In the second part of this work we investigated the effects of BN administration to patient with chronic lead intoxication. For this purpose, a phase II double-blind case-controlled randomized  clinical trial has been performed with 40 patients (manual workers in radio industry, printing, glass ad electric charge battery manufacturing and liquidators from Chernobyl atomic power station), having lead blood content about 0.4-0.6 mg/L. Experimental group was divided into 3 subgroups: the patients of Group A were given 3 g BN a day, the patients of Group B were given 6 g BN a day, and the patients of Group C were given 9 g BN a day. The control group patients were given 3 g sugar powder a day as a placebo.

It was found that the consumption  of BN  was not associated with any adverse effects in patients; on the contrary, the clinical conditions of most patients were improved. Chelatory effect of BN was dose-dependent: the administration of 6 g BN a day to patients of Group D significantly enhanced lead level in the blood (Table 2). while there was no change in the lead level of the, patients of Groups A and C. Interestingly, that the patients of Group B are also characterized by the diminished ALA and coproporphyrin contents, the parameters of lead-induced damage. Therefore,  this dosage seems to  be  optimal in  combating the consequences of lead intoxication.


Table 2

The effect of BN administration (6 g a day) on the lead blood level and the parameters of lead-induced damage

Pb content in the blood (μM/L) ALA(μM/g creatinin) Coproporphyrin(mM/g creatinin)
Before BN treatment 2.04±0.95 12.1±2.8 107±20
After BN treatment 3.56±0.95 9.7±2.5 80±15


Thus, we concluded  that due  to its chelalory and  antioxidant activity, BN  is capable of mobilizing  lead  from   the  organism’s   storage,   removing  it  from   the  organism,  and improving impaired porphyrin metaholism of patients with lead intoxication.



  1. Osato   J.A.,   Afanas’ev  I.B.,   Cheremisina  Z.P.,   Suslova T.D.,  Abramova   N.E., Mikhalchik E.V., Deeva I.B., Santiago L.A., Korkina L.G. : Bio-normalizer as a modulator of   phagocytosis   and   free   radical   production   by  murine   inflamed   neutrophils   and macrophages.  Phys.Chem.Biol.Med.  2,  1995:  87-95.  -2.  Osato  J.A.,  Korkina  L.G., Mikhal’chik  E.,  Afanas’ev I.B. : Effect of Bio-normalizer,  a  natural  Japanee   food supplement,  on oxygen radical and  tumor  necrosis factor  production  by monocytes  and macrophages. In Packer  L., Traber  M.G.,  Xin  W. (Eds): Molecular  Mechanisms  and Health Effects, AOCS Press, 1996: 109-116. -3. Lewis C.P.L., Demedts M., Nemery B.: Indices of oxidative stress in hamster lung following exposure to cobalt (II) ions: In vivo and In vitro studies. Am.J.Respir.Cell Moi.Biol. 5, 1991: 163-169.