presented by duquesne marilyn belgium presented by duquesne marilyn belgium 3 rd international...

Presented ByDuquesne MarilynBelgium

3rd International Conference and Exhibition on Traditional & Alternative Medicine

August 03-05, 2015 Birmingham, UK

In Association with

Introduction

Purpose of the

research

Method

Results

Conclusion

Herbal remedies : Natural but not unhazardous

Study cases of Aristolochic Acid intoxication

Introduction

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Introduction 2

Common term

Mixture of :

Aristolochic Acid I (AAI) Aristolochic Acid II (AAII)

&

nitrophenantrene carboxylic acid derivatives

What’s Aristolochic Acid ?

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Introduction 3

Produced by :

Aristolochia Asarum

www.fr-academic.ru

Bragantia

www.afleurdepau.com etc.usf.edu

What’s Aristolochic Acid ?

Aristolochiaceae

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Introduction 4

found in :

>> >

What’s Aristolochic Acid ?Medicinal properties of Aristolochia

Rhizomes : - Gastric stimulant- Fevers treatment- Malaria

Roots:- Type 2 diabetes- Snakebites

Leaves : - Skin disorders- Dyspepsia - Leishmania

Stems:- Anti-inflammatory- Diuretic- Circulation stimulating

Fruits : - Respiratory diseases- Skin wounds- Malaria

Seeds:- Biliousness- Abdominal pain

Flowers : - Diuretic- Hypoglycaemic effect

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

Tubular atrophy

Interstitial fibrosis

Low molecular weight proteinuria

I) Nephrotoxicity

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

Aristolochic acid I/IIAristolactam nitronium ion

R

NQO1CYP1A1CYP1A2

NQO1COXCPR

R

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Introduction 9

II) Carcinogenicity

Properties of AA-DNA adducts

Cortex

Medulla

Localisation: Responsible for:

TP53

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Introduction 10

TP53

Histochemistry

Genomic Proteomic

Metabonomic

α-1µglob.β-2µglob.

α-2 glycoprot.

INVASIVE METHOD

+

- NON INVASIVEMETHOD

INVASIVE METHOD

- NON INVASIVEMETHOD

- EARLY !!

- LATE

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Introduction 11

Balkan Endemic Nephropathy (BEN)

Chronic Kidney disease

13

Pr. Grollman (thepoisongarden.co.uk)

Etiology: Dietary intoxication

Diseased or suspects : 25000

At risks : >100000

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Introduction 12

Purpose of research

Metabonomic

Mechanism of toxicity

Exposure

AbsorptionDistributionmetabolism

Primary target

Target cell damage

Target organ

Damage to organism

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication; Purpose of research 13

BEN

Metabonomic procedure

Method :

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication; Method 14

What’s Metabonomic ?

Metabolites portrait

Mass spectro-metry

NMR

Temporal changes

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication; Method 15

Samples collection

& preparation

4°C

Antibacterial

Phosphate buffer

External Reference (TSP)

Metabonomic procedure

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication; Method 16

Metabonomic procedure

Spectroscopic analysis

NMR

Proton

electron

Magnetic field

Z

Y

X

O

Bo

RFTime

Intensity

Fourier transform

Frequency

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication; Method 17

Spectroscopic analysis

Spectrum treatment &Multivariate analysis

Metabonomic procedure

PC1

PC2

SCORES plot

PC1

PC23.40

2.501.24

3.80

5.703.24 0.80

Observations Variables

PC1

LOADINGS plot

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication; Method 18

Spectroscopic analysis

Spectrum treatment &Multivariate analysis

Biological interpretation

Metabonomic procedure

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication; Method 19

Acute toxicity Chronic toxicity

- Diseased - Suspects- At risks

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results 20

Results

Physiological data AAI AAI/AAII AAI AAI/AAII AAI AAI/AAII

Body Weight

Food intake

Water intake

Urinary volume

+ +

N.D

+

+ +

- -

-

+

+ +

+

N.D

+

+ +

- - -

- -

-

N.D

-

-

- - +

+ +

- - - -

- - -

- -

+

Clinical chemistry AAI AAI/AAII AAI AAI/AAII AAI AAI/AAII

BUN

Sr. Creatinine

Albumin

≠

≠

≠

+ + +

++≠

+ +

+≠

+ + + +

+++≠

+ +

+≠

+ + + +

+++≠

50mg/kg 75mg/kg 100mg/kg

↘

Polyuria

↗↗ ↗

75 100

AAII

- - - - -

- - - -

- - - - -

- - - -

+ + + +

+++≠

+ + + +

+ + +≠

Toxicity : AAI < AAI/AAII ? AAII ?

Results AA acute toxicity

I) Physiological Parameters

AAI AAI/AAII AAI AAI/AAII AAI AAI/AAII

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results : Acute toxicity (rats) 21

A 50 mg/kg AAIB 75 mg/kg AAIC 100 mg/kg AAID 50 mg/kg AAI/AAIIE 75 mg/kg AAI/AAIIF 100 mg/kg AAI/AAII

Results AA acute toxicity

I) Metabonomic results

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results : Acute toxicity (rats) 22

Hippurate

GlucoseBetaine

Osmoprotectants Lactate & acetate

Results AA acute toxicity

I) Metabonomic results

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results : Acute toxicity (rats) 23

↘ Hippurate

↗ Glucose

GlomerulusProximal

tubuleDistaltubule

↘ Cycle Krebs components

↗ Osmoprotectants

↗ Betaine ↗ Lactate & acetate

Results AA acute toxicity

Biological interpretation

Cortex

Medulla

↗↗ Bun↗ Ser. Creat.

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results : Acute toxicity (rats) 24

↗ Nitrogenous waste in blood↗ Osmoprotectants in urine

Plasma

↘ Reabs. Glucose

AAHippurate

Results AA acute toxicity

Biological interpretation

OAT1 ↘ Hippurate

Deficient cycle Krebs

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results : Acute toxicity (rats) 25

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

Results AA chronic toxicity

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results : Chronic toxicity ( Humans) 26

Creatinine filtration : VHippurate secretion : VGlucose reabsorption : V

Creatinine filtration : XHippurate secretion: XGlucose reabsorption : X

Metabonomic profile : Healthy

Creatinine filtration : XHippurate secretion : XGlucose reabsorption : V

Metabonomic profile:Moderate renal injury

Metabonomic profile:Severe renal injury

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

Results AA chronic toxicity

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results : Chronic toxicity ( Humans) 27

HealthyModerate renal injurySevere renal injury

Comparison of the different metabonomic profiles : healthy , moderale and severe renal injury

Renal damage severity

Results AA chronic toxicity

Herbal remedies : natural but not unhazardous. Study cases of AA intoxication Results : Chronic toxicity ( Humans) 28

Perspectives & Conclusions

Acute toxicity model

Chronic toxicity model

In process

Metabolites identification : early & easily biomarkers ( ↗ Glucose, betaine, osmoprotectants ; ↘ hippurate, cycle Krebs components )

New samples

BEN

Biological hypotheses ( renal functions, transport injuries,…)

Perspectives & Conclusion

Metabonomic

- NON INVASIVEMETHOD

- EARLY Biomarkers

Lab. Human Biology & ToxicologyUMONS (Belgium)

High possibility of human AA intoxication cases !!

Preventive action

Duquesne MarilynPhD student

Human Biology & Toxicology laboratoryUniversity of Mons(Belgium)

Profile : Familial history

Proteinuria : ACR, alpha-1

Azotemia (serum

creatinine)

anemia 15 years in EN

Diseased v v v v

v v v v

v v v v

Suspects v v

v v

At risks v

Others : x x x x

http://biology-igcse.weebly.com/dialysis.htmlhttp://www.nuhospitals.com/renal-kidney-transplant.html

https://sites.google.com/a/montessori.edu.co/biology-projects/dna-vocabulary-2/_draft_post-12http://www.medicinenet.com/kidney_infection/article.htm

http://carcin.oxfordjournals.org/content/24/10/1695/F1.large.jpghttp://www.google.be/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=&

url=http%3A%2F%2Fwww.ayurvedindia.in%2FOthers%2FLiver-Disorders%2F961&ei=inE3VeL7GdbsaMyxgYAE&psig=AFQjCNFvJDXErveYz2GG_VmWaQIoHQKqeA&ust=1429783307048964

https://www.flickr.com/photos/14179358@N05/3678083407/

WWW.stjude.comtutocours.fr

www.michigananimalhospital.comwww.vcahospitals.com

www.riversideonline.comwww.mhhe.com

hyperphysics.phy-astr.gsu.edu

Herbal remedies : natural but unhazardous. Study cases of AA intoxication ; Introduction

Traditional Medicine-2016Website: http://traditionalmedicine.conferenceseries.com/

Meet the eminent gathering once again at

Traditional Medicine-2016London, UK

October 03-05, 2016