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An in vitro and in vivo investigation of the anti-inflammatory

properties of coal derived humates.


E.J. van Rensburg, G. Joonй and A.D. Cromarty, Department of Pharmacology, Faculty of Health Sciences, University of Pretoria, South Afr

INTRODUCTION: RESULTS: Measurement of CR3 expression:


mic substances occur widely in nature. Contact hypersensitivity  Both potasium humate products caused a dose related
e therapeutic properties of humates have  Both the prednisilone and the leonardite derived humate product inhibition of CR3 expression by stimulated, but not resting
escribed as antibacterial, antitoxic, anti- caused a significant (P < 0.5) decrease in ear swelling at 3h as well as neutrophils that was significant (p< 0.05) at 10µg/ml and higher
genic, anti-arthritic, anti-allergic, 24 and 48h (Figure 2). Prednisilone proved to be superior to the (Figure 4).
omodulatory and anti-inflammatory. leonardite humate whereas the humate product derived from
ious in vivo studies have been done to bituminous coal had no significant effect on ear swelling. Figure 4
strate the anti-inflammatory activity of crude  No signs of toxicity were observed during the 7 days of treatment The effects of a 15 min treatment with various concentrations of
ts prepared from humus matter such as peat, with the humate products. However, the rats on prednisilone lost bituminous coal derived and the leonardite derived humate
and sapropel. weight compared to the control group (Figure 3). products on the expression of CR3 on resting and PMA-stimulat
documentation could be found concerning the human neutrophils determined by flow cytometry.
of coal-derived humates on inflammatory Figure 1
ns. These preparations, especially products Control rat after DNFB challenge.
d from brown coals, contain much higher
of high quality humate, are easier to produce
controlled conditions and are more available
re.
as been documented that agents that block Figure 2
hesion molecule, CR3, expressed on the Difference in ear thickness between left and right ears of DNFB
e of activated neutrophils, are beneficial in the challenged rats either without treatment or after a daily treatment for
ent of inflammation by inhibiting recruitment of one week, administered by gavage, of one of the following; 61mg/Kg
ytes into tissues. bituminous humate; 61mg/Kg leonardite humate; 1mg/Kg prednisilone.
The three columns represent the differences in ear thickness at 3
AIM OF STUDY: hours, 24 hours and 48 hours post challenge. Significant differences
ermine: compared to untreated controls.
ether oral treatment with potassium humate * p< 0.05; ** p< 0.01
period of a week could decrease the
matory effects caused by a contact
ensitivity reaction in vivo and to compare the
with prednisilone.
effects humates have on the expression of
y activated neutrophils. Figure 3
METHODS: The changes in the body mass of rats after one week of treatment with DISCUSSION:
e: 61 mg /Kg/day bituminous coal derived and leonardite derived humate  The antiinflammatory properties of oral potassium huma
umate products were dissolved in water and products or 1 mg/Kg/day prednisilone compared to untreated controls. derived from leonardite compared favourably with
ested. * p< 0.05; ** p< 0.01 prednisilone
e produced from brown coal (leonardite)  Both of the humate products inhibited CR3 expression on
ntains 92% soluble humate and fulvate). activated neutrophils.
 The difference in the activities between the two humate
e synthesized from bituminous coal (contains products in the contact hypersensitivity model needs to be furthe
investigated.
% soluble humate).
 An over-expression of CR3 is associated with the production
a multitude of cytokines, reactive oxygen, nitrogen intermediates
ct hypersensitivity: and proteolytic enzymes that can cause tissue injury and lead to
s experiment was done at the University of inflammatory conditions.
a Biomedical Research Centre.  No signs of toxicity was observed with the two humate
female Sprague Dawley rats of 8 to 10 weeks, products.
into four groups of 15 rats each.  Potassium humate has been proven to be safe in human
day 0, the rats were weighed and sensitized whereas prednisilone is associated with serious side effect
nting the shaved abdomen with 400µl of a
olution of 2,4-dinitro-fluorobenzene (DNFB) in
e:olive oil (4:1) and placed on one of the
ng oral treatments: (I) water only (2) leonardite CONCLUSION:
e (61mg soluble humate /Kg) (3) bituminous
umate (61mg soluble humate /Kg) (4)
silone (1mg/Kg). The identification of a relative nontoxic
day 6, the rats were challenged on the right compound such as potassium humate tha
application to upper surface of the ear, of exerts its anti-inflammatory properties via th
a 0.5% solution of DNFB in acetone:olive oil
Figure 1). blocking of an adhesion molecule that play
ee hours after challenge both ears were an key role in inflammation, is therefore an
red with an engineering caliper across the ear exciting finding and merits further evaluatio
stance of 3mm from the tip. Each ear was
red 3 times and the average thickness in the treatment of patients suffering from
d. The measurements of the ears were inflammatory conditions.
ed after 24 and 48 hours.

rement of CR3 expression:


uspension of neutrophils was treated with
s concentrations of the humate products for 15
37°C, stimulated with 100ng/ml PMA and
ted for a further 15 min. CR3 quantitation was
y flow cytometry analysis on a Coulter Epics
C flow cytometer using an anti-CD11b FITC
lonal antibody (Beckman Coulter, Pallo Alto,
nia).

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