HirokoSegawaDepartmentofMolecularNutri7on,Ins7tuteof
BiomedicalSciences,TokushimaUniversityGraduateSchool
KDIGOCKD-MBDImplementa7onSummitKeioPlazaHotelTokyo,Japan
InVivoResponsesofPhosphorus-basedFoodwithDifferent
FormsinMice
• Dietaryphosphorusmanagementisimportantfornotonly
renalpa7entsbutalsohealthypeoplewithnormalkidney
func7on,becausephosphorusintakeisassociatedwith
osteoporo7cfractures,leOventricularmass,andallcause
mortality,eveninhealthypopula7ons.
• Thedailyphosphateintakevarieswithageandgender.
• Onaglobalscale,therearequitesignificantdifferences
indietcomposi7onstobeconsidered.
Grains19.02%
Potates1.46%
Sugars&sweeteners0.00%
Soy7.07% Nuts,seeds
0.83%
Vegetables7.90%
Fruits1.66%
Mushroom1.46%Algae
0.62%
Seafood16.74%Meat,poultry
11.43%
Eggs6.55%
Dairyproducts12.79%
Oils0.00%
Bakery2.39%
Beverages2.39%
Seasoning7.38%
Supplementarynutri7on0.31%
Cited; 2010 National Health and Nutrition Survey, Table 9
No.1
No.3
No.2
Sources of phosphorus in Japanese food supply, 2010
Male; 4153, Female; 4662
0
200
400
600
800
1000
1200
1400
1600
2012 2013 2014 2015 2016
Male(>20yearsold)Female(>20yearsold)
Phosp
horu
s (m
g /da
y )
Na>onalHealthandNutri>onSurveyJapan
AnnualChangesintheMeanPhosphorusIntake(aged20yearsandover)(2012to2016)
Cited; 2012 - 2016 National Health and Nutrition Survey,
0
200
400
600
800
1000
1200
1400
1600
2012 2013 2014 2015 2016
Male(>20yearsold)Female(>20yearsold)
Phosp
horu
s (m
g /da
y )
Na>onalHealthandNutri>onSurveyJapan
AnnualChangesintheMeanPhosphorusIntake(aged20yearsandover)(2012to2016)
AdequateIntake;1000mg/day
DietaryReferenceIntakesforJapanese(2015)
AdequateIntake; 800mg/day
Cited; 2012 - 2016 National Health and Nutrition Survey,
• Assessmentofdietaryphosphorusiscomplicatedby
inaccuraciesinmostnutrientcomposi7ondatabasesdueto
theextensiveandvarieduseofphosphorus-containing
foodaddi7ves.
• Intakefromprocessedfoodsispar7cularlyobscuredbythe
factthatnutri7onallabelsarenotrequiredtoreportthe
phosphoruscontentoffood.
• Dietaryphosphorusintakecouldbees7matedfrom24-hurine
collec7onbyurinaryphosphorusexcre7ondividedbyphosphorus
absorp7onrate(65%).
• Phosphorusabsorp7onratesinpreviousstudiesvarywidelypossiblyduetofactorsassociatedwithitsregula7on,suchasthetypeof
phosphorus,othermineralsandinterindividualvaria7oninprior
nutrientstatusortransit7me.
• Dietaryrecords(DR)areatypeofself-reporteddietaryassessment
method.
• Althoughwidelyused,DRhaveinherentproblemsandrequirement
forpar7cipantliteracy.
• Theseproblemscannotbeavoidedinna7onalrepresenta7ve
surveys.
0
200
400
600
800
1000
1200
1400
1600
2012 2013 2014 2015 2016
Male(>20yearsold)Female(>20yearsold)
AnnualChangesintheMeanPhosphorusIntake
(aged20yearsandover)(2012to2016)
Studydesign• Thisobserva7onalstudywasbasedondatafromacrosssec7onal
mul7centersurveyconductedinFebruaryandMarch2013.
• Thesurveytargetedhealthymenandwomenaged20–69years
livingthroughoutJapan.
• Thetotalnumberofpar7cipantswas395menand396women.
ShinozakiN.etal.EuropeanJournalofClinicalNutri7on(2018)72:517–525
• Bodyweightandweremeasured
• BMIwascalculated.
• Informa7ononage,educa7on,medicalhistory,currentmedica7onuse,
physicalac7vity,andsmoking
• Es7matesofphysicalac7vitywerecalculated.
• Dietaryintakewasassessedby4-daysemi-weighteddietaryrecords.
• Urinecollec7onswereperformedontwonon-consecu7vedays.
• Atotalof161menand161womenaged20–69yearscompleteda4-day
dietaryrecordsandtwo24-hurinecollec7on.• Allanalyseswereperformedusingthesta7s7calsoOwarepackageSAS
version9.4.• Two-sidedP-values<0.05wereconsideredsta7s7callysignificant.
ShinozakiN.etal.EuropeanJournalofClinicalNutri7on(2018)72:517–525
Table1Characteris7csof322apparentlyhealthymenandwomenpar7cipantsaged20–69years,2013
ShinozakiN.etal.EuropeanJournalofClinicalNutri7on(2018)72:517–525
Men WomenbyDietaryrecord 1176 1021
byUC 1393 1082
020040060080010001200140016001800
Phosphorusintakees7matedbyUrineCollec7onwashigherthanthates7matedbyDietaryrecord
ShinozakiN.etal.EuropeanJournalofClinicalNutri7on(2018)72:517–525
P<0.0001
P=0.008*
*
Dietaryph
osph
orusintake
(mg/day)
n=161
n=161
200 60
Men WomenbyDietaryrecord 1176 1021
byUC 1393 1082
020040060080010001200140016001800
P<0.0001
P=0.008�*�
*�
Dietaryph
osph
orusintake
(mg/day)�
AdequateIntake(AI);1000mg/day
DietaryReferenceIntakesforJapanese(2015)
AdequateIntake(AI); 800mg/day
PhosphorusintakewassufficientlyhigherthanAdequateintakeinDRI.
ShinozakiN.etal.EuropeanJournalofClinicalNutri7on(2018)72:517–525
200 60
Mono-phosphateex)DisodiumhydrogenphosphateTrisodiumhydrogenphosphate
Poly-phosphatex)Potassiumpolyphosphate
Calciumdihydrogenpyrophosphate
Phosphatecontainingfood
Organicphosphate Inorganicphosphate
Foodaddi?ves
Orthophosphoricacid Pyrophosphoricacid triphosphate
NaturalsourcesAnimals,vegetables
Dietaryphosphorus
Organicphosphorus Inorganicphosphorus
AnimalproteinVegetableprotein
Foodaddi7ves
Mono-phosphatesalts Poly-phosphatesalts
• EnhanceFlavor• ImproveColor• Extendtheshelflife• Moisturereten>on• Improvetexture• Smoothness• Binding• etc.
Phosphorusaddi7vesareusedforthesefunc7ons……
Infla>ng
Moisturereten>on
Binding
Improvetexture
ü DicalciumPhosphateü Hexametaphosphateü MonocalciumPhosphateü PhosphoricAcidü Ca,Na,orK-polyphosphate
CommonPhosphorusAddi7ves
processedmeat
3Na+
?
HPO42-
Pathways for phosphate absorption in the small intestine
Basal
Apical
Inorganic monophosphate
Paracellular pathway
Transcellular pathway
Absorption system
Intestinal epithelial cell
Facilitatetransport
Transporter
Dietary Pi Digestion systemProtein
polyphosphate
3Na+
?
HPO42-
Basal
Apical
Inorganic mono phosphate
Transcellular pathway
Absorption system
NaPi2b
Transcellular pathway
2Na+
Transcellular pathway
?
PiT1/2
Paracellular transport is a major system of intestinal Pi absorption
DietaryPi
Npt2b
HPO42-
HPO42-
HPO42-
?
3Na+
Pit-1NHE3
Na+
H+
Na+ H2PO4-
P P P
P
P
P PP P
P
P
P
P
P
P
PP
P P
P
??
?
ü apicalsideiIntes7ne,colon,kidney….)ü Na+,H+exchangeü playsamajorroleintransepithelial
absorp7onofNa+andwaterü modulatestheabsorp7onofother
nutrients
NHE3/SLC9A3(Sodium+/hydrogen
exchanger3) Na+
H+
NHE3
H+
PepT1
Di,tripep>des
H+ pep>des Na+
Na/KATPase
K+
aminoacids Aminoacidtransporters
Lumen
Bloodaminoacids
Na+
DietaryPi
Npt2b
HPO42-
HPO42-
HPO42-
?
3Na+
Pit-1NHE3
Na+
H+
Na+ H2PO4-
P P P
P
P
P PP P
P
P
P
P
P
P
PP
P P
P
claudins
HPO42-
claudinsParacellularPipathway
? ?
?
?
Transcellular and Paracellular Pi transport
ü JNutrSciVitaminol(Tokyo).1999Oct;45(5):595-608.Comparisonofvariousphosphatesaltsasthedietaryphosphorussourceonnephrocalcinosisandkidneyfunc>oninrats.MatsuzakiH1,KikuchiT,KajitaY,MasuyamaR,UeharaM,GotoS,SuzukiK.
ü BiosciBiotechnolBiochem.2001Apr;65(4):928-34.Greatereffectofdietarypotassiumtripolyphosphatethanofpotassiumdihydrogenphosphateonthenephrocalcinosisandproximaltubularfunc>oninfemaleratsfromtheintakeofahigh-phosphorusdiet.MatsuzakiH1,MasuyamaR,UeharaM,NakamuraK,SuzukiK.
Mono-phosphateex)DisodiumhydrogenphosphateTrisodiumhydrogenphosphate
Poly-phosphatex)Potassiumpolyphosphate
Calciumdihydrogenpyrophosphate
Phosphatecontainingfood
Organicphosphate Inorganicphosphate
Foodaddi?ves
Orthophosphoricacid Pyrophosphoricacid triphosphate
NaturalsourcesAnimals,vegetables
Polyphosphatesalthavemoreharmfuleffectsthanthoseofmonophosphatesaltonbonephysiologyandrenalfunc7on.
27? Basal
Apical
Inorganic monophosphate
Dietary Pi
Poly-P
?
transporter
Gastrointestinal phosphate handling
Intestinal epithelial cell
Pi Pi
?
Pi
Pi
Pi
Mono-P
Poly-P
Pi
Pi
OrganicInorganic
Mono-P
Pi
Pi
Pi Pi
AnimalC57BL6J,5weeksofage♂ TestdietGroup1. Low Pi diet 2. Control Pi diet 3. Monophosphate HP 4. Polyphosphate HP
AnimalsandDiet
Pi Pi
Pi
Pi Pi
Mono-PHighphosphatediet
Pi
Poly-PHighphosphatediet
Diet 100 g �g�LP CP HP1 HP2
AIN93G modified Mix 59.5 59.5 59.5 59.5Egg White 20 20 20 20
AIN93G modified Mineral Mix 1.5645 1.56451.56451.5645
Sucrose 10.4371 7.88715.25214.7421CaCO3 1.4984 1.49841.49841.4984
KH2PO4 or K5P3O10 0 2.55 5.185 5.695Soy Bean Oil 7 7 7 7
Total 100 100 100 100
MonoHP PolyHP
Timeschedule
0 7 28 32 33 3435�
�days�
Metaboliccage
Sacrificed
5weeks
Start5weeksofage 10weeksofage
Pi Pi
Pi
Pi Pi
Mono-PHighphosphatediet
Pi
Poly-PHighphosphatediet
1. Low Pi diet 2. Control Pi diet 3. Monophosphate HP 4. Polyphosphate HP
Testdiet
0
5
10
15
20
25
30
35
40
LP CP HP1 HP20
1
2
3
4
5
6
7
LP CP HP1 HP2
0
2
4
6
8
10
LP CP HP1 HP2
a
a,b
0
1
2
3
4
5
LP CP HP1 HP2
a,b
a aa
a
aa
aa a
Pla
sma
Pi (
mg/
dl)
Pla
sma
Ca
(mg/
dl)
Urin
e P
i (m
g/da
y)
Urin
e C
a(m
g/da
y)
0
50
100
150
200
250
300
LP HP1 HP2
FGF2
3 (p
g/m
L)
0 20 40 60 80
100 120 140 160 180
LP HP1 HP2
a aa
PTH
(pg/
mL)
MonoHP
PolyHP
MonoHP
PolyHP
MonoHP
PolyHP
MonoHP
PolyHP
MonoHP
PolyHP
MonoHP
PolyHP
LP CP MonoHP PolyHP
LP CP HP1 HP2
Massontrichromestain
VonKossaCalciumstain
Polyphosphatedietinducedrenalfibrosisandcalcifica7on.
MonoHP PolyHP
Whatisthedifferencebetweenmono-Pdietandpoly-Pdiet?
Pi Pi
Pi
Pi Pi
Mono-PHighphosphatediet
Pi
Poly-PHighphosphatediet
Pi Pi Pi
Proxim
alintes7ne
ATP(n
M)
ATP:SubstrateforIAPIntes7nalalkalinephosphatase
(IAP)
0
0.2
0.4
0.6
0.8
1
1.2
HP1 HP2
0
0.5
1
1.5
2
2.5
HP1 HP2
Proxim
alintes7ne
p-NP
(mmol/m
gprotein/min)
HP1 HP2HP1 HP2
* *
MonoHP
PolyHP
MonoHP
PolyHP
Iden>fica>onofspecifictargetsforthegutmucosaldefensefactorintes>nalalkalinephosphataseKathrynT.Chen,1,2MadhuS.Malo,1AngelaK.Moss,1SkyeZeller,3PaulJohnson,3FarzadEbrahimi,1GolamMostafa,1SayedaN.Alam,1SundaramRamasamy,1H.ShawWarren,3ElizabethL.Hohmann,3andRichardA.Hodin1
Microbiota–hostinterac7onandmodula7onbydietarynutrients.
Uremictoxin
Nutrients2017,9,489;doi:10.3390/nu9050489modified
Processedbymicrobiota
Growth:Prolifera7onofmicrobiotadependenton
specificnutrients
Absorbedinthegut
Uptakebymicrobiota
Processedbyhost
Growth/Tissueandfunc7onmaintenance
L-carni7ne,choline/phospha7dylcholine,tryptophanandtyrosine…Pi..
UremictoxinOr
Uremictoxinpre
Nutrients
Hostwaste Microbialwaste
MonophosphateHP PolyphosphateHP
Pi Pi
PiPiATP
IAP
IAP
IAPIAP
degrada7on
microbiota microbiota
ATP
ATPATP
種類分布
Type,Numberchange
BloodPiHigh BloodPiHigh
Pi Pi
Hypothesis
Uremictoxin