3rd International Conference and Exhibition on
Traditional & Alternative Medicine
August 03-05, 2015 Birmingham, UK
In Association with
Presented By
Duquesne Marilyn
Belgium
Herbal remedies : Natural but not
unhazardous
Study cases of Aristolochic Acid intoxication
Introduction
Purpose of the research
Method
Results
Conclusion
Introduction
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Introduction
2
What’s Aristolochic Acid ?
 Common term
 Mixture of :
&
Aristolochic Acid I (AAI)
Aristolochic Acid II (AAII)
nitrophenantrene carboxylic acid derivatives
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Introduction
3
What’s Aristolochic Acid ?
 Produced by :
Aristolochiaceae
Aristolochia
Asarum
Bragantia
www.fr-academic.ru
www.afleurdepau.com
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
etc.usf.edu
Introduction
4
What’sproperties
Aristolochic
Acid ?
Medicinal
of Aristolochia
 found in :
>
>
>
Rhizomes :
Fruits :
- Gastric stimulant
- Fevers treatment
- Malaria
- Respiratory diseases
- Skin wounds
- Malaria
Roots:
- Type 2 diabetes
- Snakebites
Leaves :
- Skin disorders
- Dyspepsia
- Leishmania
Stems:
Seeds:
Flowers :
- Diuretic
- Hypoglycaemic effect
- Biliousness
- Abdominal pain
- Anti-inflammatory
- Diuretic
- Circulation stimulating
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Introduction
5
Aristolochic acid
toxicity
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Introduction
6
I) Nephrotoxicity
Interstitial fibrosis
Tubular atrophy
Low molecular weight
proteinuria
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Introduction
7
I) Nephrotoxicity
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Introduction
8
II) Carcinogenicity
 Induced by metabolisation of AA
NQO1
CYP1A1
CYP1A2
R
Aristolochic acid I/II
NQO1
COX
CPR
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
R
Aristolactam nitronium ion
Introduction
9
II) Carcinogenicity
 Properties of AA-DNA adducts
Localisation:
Responsible for:
Cortex
TP53
Medulla
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Introduction
10
+
Metabonomic
- NON INVASIVE
METHOD
Histochemistry
INVASIVE
METHOD
INVASIVE
METHOD
TP53
- EARLY !!
α-1µglob.
- NON
INVASIVE
β-2µglob.
METHOD
α-2
glycoprot.
- LATE
Genomic
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Proteomic
Introduction
11
Balkan Endemic Nephropathy (BEN)



Chronic Kidney disease
Diseased or suspects : 25000
At risks : >100000

Etiology:

Dietary intoxication
Pr. Grollman (thepoisongarden.co.uk)
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Introduction
13
12
Purpose of research
BEN
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication;
Exposure
Absorption
Distribution
metabolism
Damage to organism
Mechanism
of toxicity
Primary target
Target organ
Target cell damage
Purpose of research
13
Method :
Metabonomic procedure
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication;
Method
14
What’s Metabonomic ?
Metabolites portrait
Temporal changes
Mass spectrometry
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication;
NMR
Method
15
Metabonomic procedure
Phosphate buffer
Samples collection
&
preparation
External
Reference (TSP)
Antibacterial
4°C
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication;
Method
16
Metabonomic procedure
Magnetic field
electron
Proton
Spectroscopic analysis
NMR
Z
Bo
Intensity
RF
O
Frequency
Time
Y
Fourier
transform
X
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication;
Method
17
Metabonomic procedure
Spectroscopic analysis
SCORES plot
LOADINGS plot
PC2
3.40
2.50
3.80
PC2
PC1
1.24
PC1
PC1
5.70
3.24 0.80
Observations
Variables
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication;
Spectrum treatment &
Multivariate analysis
Method
18
Metabonomic procedure
Spectroscopic analysis
Biological interpretation
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication;
Spectrum treatment &
Multivariate analysis
Method
19
Results
Acute toxicity
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Chronic toxicity
- Diseased
- Suspects
- At risks
Results
20
Results
AA acute toxicity
I) Physiological Parameters
50mg/kg
↘
𝑓 𝑎𝑔𝑒𝑛𝑡 &𝑑𝑜𝑠𝑒
Polyuria
↗↗
↗
75mg/kg
100mg/kg
75
100
Physiological
data
AAI
AAI/AAII
AAI
AAI/AAII
AAI
AAI/AAII
Body Weight
++
--
+
---
-
----
-----
-----
Food intake
N.D
-
N.D
--
-
---
----
----
Water intake
+
+
+
-
--+
--
Urinary volume
++
++
++
N.D
++
+
Clinical
chemistry
AAI
AAI/AAII
AAI
AAI/AAII
AAI
AAI/AAII
BUN
≠
+++
++
++++
++
++++
++++
++++
Sr. Creatinine
≠
++
+
+++
+
+++
+++
Albumin
≠
≠
≠
≠
≠
≠
≠
+++
≠
Toxicity : AAI < AAI/AAII
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
AAII
 ? AAII ?
Results : Acute toxicity (rats)
21
Results
AA acute toxicity
I) Metabonomic results
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
A 50 mg/kg AAI
B 75 mg/kg AAI
C 100 mg/kg AAI
D 50 mg/kg AAI/AAII
E 75 mg/kg AAI/AAII
F 100 mg/kg AAI/AAII
Results : Acute toxicity (rats)
22
Results
AA acute toxicity
I) Metabonomic results
Hippurate
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Glucose
Betaine
Osmoprotectants
Lactate & acetate
Results : Acute toxicity (rats)
23
Results
AA acute toxicity
Biological interpretation
↗↗ Bun
↘ Hippurate
↗ Glucose
↗ Betaine
↗ Lactate & acetate
↗ Osmoprotectants
↗ Ser. Creat.
↘ Cycle Krebs components
Cortex
Glomerulus
Proximal
tubule
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Distal
tubule
Medulla
Results : Acute toxicity (rats)
24
AA acute toxicity
Results
Biological interpretation
AA
Hippurate
OAT1
↘ Hippurate
↘ Reabs. Glucose
Plasma
Deficient cycle Krebs
↗ Nitrogenous waste in blood
↗ Osmoprotectants in urine
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Results : Acute toxicity (rats)
25
Results
AA chronic toxicity
Metabonomic profile reflecting the severity of renal damage in
proximal tubules and/or interstitial tissue:
Psihogios, N ; Kalaitzidis, R ; Dimou, S ; Seferiadis,K ; Siamopoulos, K ; and Bairaktari, E. Evaluation of Tubulointerstitial lesions
severity in patients with glomerulonephritides : An NMR-based metabonomic study. J.Proteome Res. 2007, 6, 3760-3770
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Results : Chronic toxicity ( Humans)
26
Results
AA chronic toxicity
Metabonomic comparison diseased, suspects, at risks, others , NEN with
Belgium control samples.
Metabonomics: on the Road to Detect Diagnostic Biomarkers in Endemic (Balkan) Nephropathy. Evaluation in a Retrospective Pilot Project.
Duquesne et al., J Cancer Sci Ther 2012
Creatinine filtration : X
Hippurate secretion : X
Glucose reabsorption : V
Metabonomic profile:
Moderate renal injury
Creatinine filtration : V
Hippurate secretion : V
Glucose reabsorption : V
Metabonomic profile : Healthy
Metabonomic profile:
Severe renal injury
Creatinine filtration : X
Hippurate secretion: X
Glucose reabsorption : X
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Results : Chronic toxicity ( Humans)
27
Results
AA chronic toxicity
 Comparison of the different metabonomic profiles : healthy , moderale and severe renal injury
Healthy
Moderate renal injury
Severe renal injury
Renal damage severity
Herbal remedies : natural but not unhazardous. Study cases of AA intoxication
Results : Chronic toxicity ( Humans)
28
Perspectives & Conclusions
Acute toxicity model
Metabolites identification : early & easily biomarkers
( ↗ Glucose, betaine, osmoprotectants ; ↘ hippurate, cycle Krebs components )
Biological hypotheses
( renal functions, transport injuries,…)
In process
Chronic toxicity model
BEN
New samples
Perspectives & Conclusion
Metabonomic
- NON INVASIVE
 High possibility of human AA intoxication cases !!
METHOD
- EARLY Biomarkers
Lab. Human Biology & Toxicology
UMONS (Belgium)
Preventive action
Duquesne Marilyn
PhD student
Human Biology & Toxicology laboratory
University of Mons(Belgium)
Profile :
Familial
history
Proteinuria
: ACR, alpha1
Azotemia
(serum
creatinine)
anemia
Diseased
v
v
v
v
v
v
v
Suspects
v
v
At risks
v
Others :
x
v
v
v
v
v
v
x
v
x
15 years in
EN
x
http://biology-igcse.weebly.com/dialysis.html
http://www.nuhospitals.com/renal-kidney-transplant.html
https://sites.google.com/a/montessori.edu.co/biology-projects/dna-vocabulary-2/_draft_post-12
http://www.medicinenet.com/kidney_infection/article.htm
http://carcin.oxfordjournals.org/content/24/10/1695/F1.large.jpg
http://www.google.be/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=&url=http%3A%2F%2Fww
w.ayurvedindia.in%2FOthers%2FLiverDisorders%2F961&ei=inE3VeL7GdbsaMyxgYAE&psig=AFQjCNFvJDXErveYz2GG_VmWaQIoHQKqeA&ust=1429783307048964
https://www.flickr.com/photos/14179358@N05/3678083407/
WWW.stjude.com
tutocours.fr
www.michigananimalhospital.com
www.vcahospitals.com
www.riversideonline.com
www.mhhe.com
hyperphysics.phy-astr.gsu.edu
Herbal remedies : natural but unhazardous. Study cases of AA intoxication ;
Introduction
Meet the eminent gathering once again at
Traditional Medicine-2016
London, UK
October 03-05, 2016
Traditional Medicine-2016
Website: http://traditionalmedicine.conferenceseries.com/