Glycative Stress Research
Online edition : ISSN 2188-3610
Print edition : ISSN 2188-3602
Received : October 17, 2022
Accepted : November 11, 2022
Published online : December 31, 2022
doi:10.24659/gsr.9.4_186

Original article

Antiglycative effect of ferulic acid
Masayuki Yagi 1), Chieko Sakiyama 1), Toshio Kitaba 2), Hiroo Kondo 2),
A. N. M. Mamun-Or-Rashid 1, 3), Tanzima Tarannum Lucy 1, 3), Yoshikazu Yonei 1)
1) Anti-Aging Medical Research Center and Glycative Stress Research Center,
Graduate School of Life and Medical Sciences, Doshisha University, Kyoto, Japan
2) Social welfare corporation KITABA society, Wakayama, Japan
3) Department of Environmental & Occupational Health, School of Public Health,
University of Pittsburgh, Pittsburgh, PA, USA

Abstract

Glycative stress is one of the factors that cause aging and accelerates development of various diseases, including diabetes.
Inhibition of glycative stress is referred to as anti-glycation. Anti-glycation includes suppression of postprandial hyperglycemia,
suppression of glycative reaction, and decomposition and excretion of advanced glycation endproducts (AGEs). Inhibitory
effects of AGE formation are recognized in various food materials such as herb teas, vegetables and fruits. It is reported
that these food materials include many phenolic compounds that inhibit the formation of AGEs. Ferulic acid (FA) is a kindof phenolic compounds included in grains such as rice, wheat, fruits, vegetables, and other food materials. Bran (grain), in
particular, includes a lot. FA is well known to be effective in inhibiting development of complications of diabetes or otherneurotic diseases because of its anti-glycative / anti-inflammatory action. In our body, many kinds of AGEs are produced
through many paths due to glycative stress, while there are many kinds of proteins that are glycated. On the other hand,
reducing the function of proteins due to glycation is associated with the cross-linking formation induced by proteins changing
into AGEs. The objective of this study was to evaluate the anti-diastatic effect of FA refined from rice bran. In this study,
inhibitory effects of protein glycation reaction models such as human serum albumins (HSA), collagen and elastin, protein
cross-linking formation by AGEs, and activity enhancement for oxidized protein hydrolase (OPH) were validated. As a result,it was clarified that FA inhibits the formation of fluorescent AGEs (F-AGEs) in all glycation reaction models. FA inhibited theformation of F-AGEs, pentosidine, CML, 3DG, GO, and MGO in the glycation reaction models of HSA-gulcose. In addition,
FA also inhibited the formation of dimmer and trimer caused by the glycation of lysozyme. FA increased the OPH activation
by a factor of 1.6. There is a possibility that anti-diastatication is associated with FA‘s usefulness for a variety of diseases.    

KEY WORDS: advanced glycation endproducts; AGEs, inhibition of AGE formation,		inhibition of glycated protein cross-linking, ferulic acid

Introduction
Glycative stress is one of the factors that cause speed
progression of various diseases, including aging and adultdiseases. Inhibition of glycative stress is called anti-glycation.
Inhibition of postprandial hyperglycemia occurs by forming
advanced glycation endproducts (AGEs), as well as degradationand excretion of AGEs 1, 2). Aminoguanidine (AG) has an
inhibitory effect on the formation of AGEs 3, 4). However, an
intake of AG has adverse reactions such as anemia, hepatic

disorder and vitamin B6 deficiency. On the other hand,various food materials including teas, herb teas, vegetables
and fruits are reported to have inhibitory effects on AGE
formation 5-7). Materials that form inhibitory effects on AGEs
included in these food materials are reported to be phenol
compounds such as chamaemeloside 8), luteolin 9), polymethoxy
flavonoid 10, 11), cyanidin3-O-galactoside 12), caffeoyl glucose 13),
rhodanthenone B 14), as well as chlorogenic acid 15). Ferulic
acid (FA) is a material usually included in a variety of foodmaterials such as grains like wheat, fruits, and vegetables.

Contact Address: Professor Masayuki Yagi, PhD
Anti-Aging Medical Research Center / Glycative Stress Research Center,
Graduate School of Life and Medical Sciences, Doshisha University
1-3, Tatara Miyakodani, Kyotanabe-shi, Kyoto, 610-0394 Japan
TEL & FAX : +81-774-65-6394 e-mail: myagi@mail.doshisha.ac.jp
Co-authors: Sakiyama C, csakiyam@mail.doshisha.ac.jp;
Kitaba T, t_kitaba@inforlive.com; Kondo H, KONDO@KITABAKAI.org;
Mamun-Or-Rashid ANM, anm541@pitt.edu; Lucy TT, cygf 2501@mail4.doshisha.ac.jp;
Yonei Y, yyonei@mail.doshisha.ac.jp_ 18 6 _Glycative Stress Research 2022; 9 (4): 186 -193(c) Society for Glycative Stress ResearchGlycative Stress ResearchBran, in particular, contains a lot 16 - 19). F A has an antioxidative effect, anti-inflammatory effect and anti-bacterial
effect which prevent us from suffering from development ofneurotic diseases, or Alzheimer disease 20), while inhibiting
obesity and lipid abnormality 21, 22). On the other hand, theanti-glycative effect of FA has been validated in glycation
reaction models which used bovine serum albumin (BSA) as amodel protein 23). The glycation reaction in a body is complex
due to its multi-path feature. There are various short-chain
aldehydes such as 3- deoxyglucosone (3DG), glyoxal (GO),and methylglyoxal (MGO) as glycative reaction intermediates,
and there are several kinds of AGEs formed from them 24). For
this reason, it is important to validate the inhibitory effect offormation, focusing on AGEs with different glycative reaction
paths by using multiple proteins 25). Furthermore, reductionof the protein function due to glycation is associated withcross-linking resulting from proteins becoming AGEs. In
this study, aiming at evaluating the anti-glycative effects of
FA refined from rice bran, the inhibitory effect of glycative
reaction intermediates, and glycation by using human serum
albumins (HSA), collagen, and elastin, protein cross-linking
formation caused by AGEs, and activity enhancement effects
for oxidized protein hydrolase (OPH) were validated.

Materials and methods
Reagent

Materials used were procured from the manufacturers as
bellow:
Human serum albumin (HSA), lyophilized powder, ≥ 96 %,agarose gel electrophoresis, methylglyoxal (MGO) solution
40 % in H2O; Sigma-Aldrich Japan (Meguro-ku, Tokyo, Japan)
Collagen type I, bovine skin, pepsin-solublilized; Nippi(Adachi-ku, Tokyo)
Elastin peptide ( P-elastin) ; Nippon Meat Packers, Inc(Osaka, Japan)
Lysozyme hydrochloride (from egg white) aminoguanidinehydrochloride (AG), 2,3-diaminonaphthalene (DAN), 40 %glyoxal (GO) solution; Dojindo Laboratolries (Kamimashiki,
Kumamoto, Japan)
CircuLex CML / Nε-(Carboxymethyl)Lysine) ELISA Kit;
MBL (Nagoya, Aichi, Japan)
Acetyl-alanine p-nitroanilide (AAPA); Bachem (Budendorf,
Switzerland)
Acylamino -acid releasing enzyme of oxidized proteinhydrolase (OPH); Takara Bio (Otsu, Shiga, Japan). And otherreagents (prime grade, or HPLC grade) were procured fromFuji Film Wako Pure Chemical and Nacalai Tesque (Kyoto,
Japan).

Sample solution
Ferulic acid (FA) refined from rice bran ( Tsuno Food
Industrial, Wakayama, Japan) was used as a reagent (Fig. 1).

Protein-glucose glycative reaction model
The inhibitory effect of glycative reaction was validated
by using the protein-glucose glycative reaction model,
referring to the previous report 26). The reaction solution was
produced: in 0.1 mol/L phosephate buffer (pH 7.4) including

1. Structureof ferulic acid.
Fig. 1. tructureFigureof ferulic
acid.

protein and glucose, sample solution was put by the volumeof one tenth of phosephate buffer. The density of protein
and glucose was as follows: 8.0 mg/mL HSA and 0.2 mol/L
glucose; 1.2 mg/mL collagen and 0.4 mol/L glucose; and 6
mg/mL elastin and 0.2 mol/L glucose.
To examine the glycative reaction, four liquids were prepared
as follows:
A. liquid which was made of phosphate buffer solution,
protein solution, glucose solution and sample solution ;B. liquid which was made of the A solution, but instead of
glucose solution, purified water was added ;C. liquid which was made of the A solution, but instead ofsample solution, purified water was added ;D. liquid which was made of the C solution, but instead of
glucose solution, purified water was added.
In the case of the HSA-glucose glycative reaction model,
incubation condition was set for 40 hours at 60 ℃, while inthe case of collagen or keratin-glucose glycative reaction
model, incubation was set for ten days at 60 ℃. The quantityof AGEs and glycated reaction intermediate was determinedby measuring the density of the reaction liquid after incubation.
Glycative reaction inhibitor, AG was used as a positive
control material to inhibit the glycative reaction.

Measurement of AGEs
Referring to the previous report 26), fluorescent AGEs
(F-AGEs) were measured. 200 μL glycative reaction solution
was put in a black microplate to measure fluorescent material
resulting from AGEs (excitation wavelength 370 nm / fluorescentwavelength 440 nm). Referring to the previous report 27),
glycative reaction solution and 6N hydrochloric acid was
mixed and was hydrolyzed for 18 hours at a temperatureof 105 ℃. Pentosidine was measured by using HPLC. For
CML, CircuLex CML / N the glycative reaction solutionε(Carboxymethyl) lysine ELISA Kit was used as glycative
reaction solution for the measurement.

Measurement of glycative reaction intermediates
The glycative reaction intermediates were measured
referring to the previous reports 28, 29). Glycative reaction
solution of 3DG, GO, and MGO were deproteinized using
perchloric acid. Then, DAN was added under the alkalinecondition. After the label handling, measurement was
performed using HPLC.
Calculation of inhibitory effect of glycative reaction
Inhibitory effects of glycative reaction were measured
referring to the previous report 26). Inhibitory rates of AGEs

_ 18 7 _Anti-glycative Effect of Ferulic Acidand glycative reaction intermediates (%) were calculated by
the following formula:
Inhibitory rate (%) = {1 − (A − B) / (C − D) } × 100
Then, 50 % inhibitory concentration (IC 50 ; mg/mL) was
calculated 26, 30), based on the inhibitory rate of sample solution
with three concentrations. IC 50 shows that the inhibitory
effect of glycative reaction gets stronger in accordance withIC 50 value getting smaller.

Inhibitory effects to form glycative
protein cross-linking
Inhibitory effects to form glycative protein cross-linking
were validated by using lysozyme-glucose glycative protein
cross-linking model 31, 32). The reaction solution was prepared:
in 0.1 mol/ L phosphate buffer solution (pH 7.4) including 5
mg/mL lysozyme and 0.5 mol/L glucose, sample solution was
one tenth volume.
There are four reaction solutions as follows:
A. mixed liquid with phosphate buffer solution, lysozyme
solution, glucose solution and sample solution ;B. liquid which was made of A solution, but instead ofgulucose solution, purified water was added ;C. liquid which was made of A solution, but instead ofsample solution, dissolving sample solution was added ;D. liquid which was made of C solution, but instead of
glucose solution, purified water was added ;The solutions were incubated for 40 hours at a temperatureof 60 ℃. After the reaction, they were placed in a centrifugal
filter with a 3 kDa ultrafiltration membrane (Amicon Ultra0. 5 mL centrifugal filters Ultracel-3K; Merck, Darmstadt,
Deutschland) to remove low-molecular substances. Then, thereaction solutions were placed in the 4-20 % polyacrylamide
gel electrophoresis (SDS-PAGE). After the migration, the gel
was stained by CBB Stain One (Nacalai Tesque). Migrating
image was processed by the Pharos FX System (Bio-Rad)and then strength of the band of dimmer (28.6 kDa) andtrimer (42.9 kDa) of lysozyme was analyzed using ImageJ(NIH, Maryland, USA). AG was used as a positive control
material for the inhibitory effects to form glycative protein
cross-linking. The rate of inhibiting protein cross-linking
formation was calculated following the formula below :Inhibitory rate of forming protein cross-linking (%)= {1 − (A − B) / (C − D) } × 100

p-nitroanilide (pNA) was separated. The separated volume
was measured by a spectrometry at 405 nm (S). In reference
to OPH activity measurement, water was added to the reaction
solution instead of the sample solution for measurement (R).
The rate of activation (%) for OPH was calculated by thefollowing formula:
OPH enhancement rate (%) = {(S 30 – S 0) / (R 30 – R 0) } × 10 0Here, pNA volume separated in 30 minutes just after
the Ref reaction started (0 minute) was set as 100 %.
S ; pNA concentration of reaction solution after sample
solution was added
R ; pNA concentration of Ref reaction solution
30; 30 minutes later
0 ; just after the start of reaction (0 minute)

Statistical analysis
Measurement values were shown by mean value ±standard deviation (SD). Tukey’s multiple comparison test
(Tukey’s test), or t test was used to compare measurement
values. As for the statistical analysis, results with less than
5 % risk rate were considered significant.

Results

Inhibitory effect of forming fluorescent AGEs onprotein-glucose glycative reaction models

Regarding FA’s inhibitory effect on the formation of
F-AGEs, it was proven that there is concentration dependence
in each glycative protein model of HSA, collagen, and elastin( Fig. 2 ).
Inhibitory effects to form F-AGEs of FA concentration
0.1 mg/mL was examined: the rate of inhibition was 52.7 ±1.4 % for HSA, 44.5 ± 0.8 % for collagen, and was 32.4 ±1.5 % for elastin (in height order). In the same way, AG has a
glycative reaction inhibiting effect. The rate of inhibition was
79.6 ± 0.2 % for HSA, 58.0 ± 1.4 % for elastin, and was 30.0
± 0.3 % for collagen (in height order). The rate of inhibition
to form F-AGEs in FA with reaction concentration 0.1 mg/mL
was 0.7 times higher than in the case of AG for HSA, 0.6
times higher (low) for elastin, and was 1. 5 times higher forcollagen (high). In terms of IC 50, AG was lower comparedto ferulic acid for HSA, collagen and elastin: IC 50 of ferulic
acid was 2.0 times the rate of AG for HSA, 1.2 times the rate
for collagen, and 3.3 times the rate for elastin ( Table 1 )

A-D ; strength of band of lysozyme dimmer or lysozyme
trimer, of the SDS-PAGE gel chromatic figure ofeach reaction solution

Inhibitory effect of forming AGEs and glycative
reaction intermediates in HSA-glucose glycative
reaction models

Measurement of activity enhancement effect of OPH

Inhibitory effects that form F-AGEs, pentosidine, CML,
3DG, GO, and MGO, by 0. 3 mg/mL FA in HSA-glucose
glycative reaction models were examined ( Table 2 ). FA’s
inhibitory effects to form AGEs and glycative reaction
intermediates were strong in the following order: F-AGEs,
pentosidine, GO, MGO, 3DG, and CML. On the other hand,FA’s inhibitory effect to form AGEs and glycative reaction
intermediates was lower than AG in all categories ( p < 0.05).
There was no comparison for pentosidine.

Activity reinforcement of OPH was examined: referringto the report 33), 0.1 mol/L Tris hydrochloric acid (pH 7.4), 2
mmol/ L Acetyl-alanine p-nitroanilide (AAPA), 1mU/mL OPH
and sample solution extract were mixed. One twenty-fifth
volume sample solution was added to the solution. Then
250 μL reaction solution was incubated for 30 minutes at
a temperature of 37 ℃. When OPH decomposed AAPA,_ 18 8 _Glycative Stress Research

HSA
100

COL

*

*

80

inhibition ratio (%)

Ela

*

60
40

*

20
0

0.01

- 20

0.1

1.0

0.1

AG

Ferulic acid

Fig. 2. Inhibitory activity of ferulic acid on F-AGE formation.

Results are expressed as mean ± SD, n = 3; * p < 0.05 by Tukey’s test. HSA, human serum albumin; COL, collagen; Ela,
elastin; AG, aminoguanidine; F-AGE, fluorescent advanced glycation endproduct; SD, standard deviation.

Table 1. Half inhibitory concentration (IC50) on the protein-glucose glycation model.
Model protein

Ferulic acid(mg/mL)

Aminoguanidine 1)(mg/mL)

HSA

0.093

0.046

Collagen

0.242

0.201

Elastin

0.231

0.069

1) Positive control of glycation inhibitor. IC50, 50 % inhibitory concentration; HSA, human serum albumin.

Table 2. AGE inhibition ratio on the HSA-glucose glycation model
AGEs compound

Ferulic acid(%)

Aminoguanidine(%)

Fluorescent AGEs

70.6 ± 0.4 *100.7 ± 0.1

Pentosidine

46.0 ± 0.2

---

CML

3.1 ± 1.1 *65.1 ± 1.1

3DG

19.8 ± 1.0 *86.7 ± 0. 2

GO

48.1 ± 1.5 *94.0 ± 1. 2

MGO

35.8 ± 1.6 *104.7 ± 1. 5

Sample concentration, 0. 3 mg/mL; mean ± SD, n = 3, * p < 0.05 vs aminoguanidine by Student’s t test;
Characteristic fluorescence (excitation/emission = 370/440 nm). AGE, advanced glycation endproduct;
CML, N ε- carboxymethyl lysine; 3DG, 3-deoxyglucosone; GO, glyoxal; MGO, methylglyoxal; HSA,
Human serum albumin; SD, standard deviation.

_ 18 9 _Anti-glycative Effect of Ferulic Acid

Discussion

Inhibitory effect of forming glycative protein
cross-linking
FA and AG inhibited the formation of lysozyme’s
dimmer and trimer in the sample solutions with concentrationof 0.05 ~ 5.1 mmol/L in a concentration-dependent manner( Table 3, Fig. 3 ). FA’s inhibitory rate to form protein crosslinking was compared to the case of AG. In each case,concentration was 1. 5 mmol/L. Regarding dimer, the rate was
0.2 times higher than the case of AG, while it was 0. 5 times
higher for trimer. As for the concentration of 5.1 mmol/L,both rates of dimer and trimer, were 0.5 times higher (low).

Activity enhancement effect of OPH
Regarding FA, when Ref (water) activation is 100 withsample concentration 0.4 μg/mL, OPH activity enhancement
effects by 166.2 ± 2.7 (n = 3) was recognized.

Ref

FA

0

1.5
(G) (-)

5.1
MW(G) (-) (G) (-)

Inhibitory effects on glycative reaction by FA

It is admitted that FA has an inhibitory effect on theformation of F-AGEs and CML in glycative reaction models
such as glucose, fructose, and ribose whose model protein is
BSA 23). In this study, FA inhibited the formation of F-AGEs
in glycative reaction models including HSA, collagen, andelastin as model proteins. Additionally, FA inhibited theformation of F-AGEs, pentosidine, CML, 3DG, GO, and
MGO in the HSA-gulcose glycative reaction model. These
results show a possibility that FA inhibits the glycation ofvarious proteins in body tissue and prevents the accumulationof AGEs. However, there is a difference in the strength ofeffect.
There is a possibility that an increase of bloodconcentration in 3DG, GO, and MGO consisting of aldehyde
might cause an apoptosis of vascular endothelium 34). An

AG
0.5(G) (-)

5.1(G) (-)

1.5
0.5 mmol / L(G) (-) (G) (-)

37kDa

Trimer

25kDa

Dimer

11kDa

MonomerFig. 3. Inhibitory effect of ferulic acid on cross-linking formation in the lysozyme-glucose reaction model.

5 mg/mL lysozyme were incubated at 60 °C for 40 hours. SDS-PAGE was conducted using 4-20 % acrylamide gels.
MW, molecular weight markers; Ref, incubation with 50 % ethanol; FA, incubation with 0.1 mg/mL ferulic acid
50 % ethanol solution; AG, incubation with aminoguanidine; (G), incubation with 0.5 mol/L glucose; (-), incubation
without glucose; mmol/L, sample concentration.

Table 3. Inhibitory effect of ferulic acid and aminoguanidine on cross-linking formation in the lysozyme-glucose reaction model.
Polymerized lysozyme

Trimer

Dimer

Concentration(mmol/L)

Ferulic acid

Aminoguanidine

Inhibition ratio (%)

Inhibition ratio (%)

0.5

13.1 ± 9. 0

39.4 ± 9.3 *

1.5

36.8 ± 1.1

69.6 ± 8.0 **

5.1

48.2 ± 2.1

90.3 ± 4.3 *0.5-4. 0 ± 3.7

12.4 ± 2.4 **1.5

5. 8 ± 1.8

33.6 ± 2.9 **5.1

28. 5 ± 2.0

53.9 ± 7.0 **5 mg/mL lysozyme were incubated at 60 °C for 40 hours. SDS-PAGE was conducted using 4 - 20 % acrylamide gels. Data are expressed as mean ± standard
deviation, n = 3 ; Dimer, 28.6 kDa; Trimer, 42.9kDa ; * p < 0.05, ** p < 0.01 vs AG by t test.

_ 19 0 _Glycative Stress Researchincrease of blood acetaldehyde also happens when the
postprandial glucose level elevates 35). In addition, when 3DG
increases by 100 nmol/L, risks for diabetic retinal disease andnephropathy double 28). Therefore, there is a possibility thatFA’s effect to inhibit aldehyde formation is associated with
the inhibition of blood vessel aging or diabetic complications.
There are many papers that validated FA’s influence on
diabetic complications 36). In the test where 100 mg/kg FA per
day was administered to STZ -diabetogenic rats for eightweeks, an improvement of renal functions associated with anamelioration of volume increase, sclerosis, and fibrosis ofglomerulus was recognized 37). In the test where 10 mg/kg FA
per day was administered to type 2 diabetic rats for 26 to 45
weeks, there were effects that improved diabetic renal disease
based on a reduction of glomerular basement membranethickness and an increase of glomerular volume andMesangium cells 38). According to these papers, FA’s usefulness
is considered to be primarily associated with anti-oxidative
effects or anti-inflammatory effects.
Based on the results of this study, there is a possibility
that FA’s inhibitory effect to develop diabetic complications
is associated with its inhibitory effect of glycative reaction.
FA is included in grains such as rice and wheat, fruits, andother vegetables. Wheat, barley, and corn contain 500 mg FA
per 100 g 19). The volume of FA available from a meal withthese grains is calculated as 20-50 mg 19). Inhibitory effects offorming AGEs by rice were validated using extracts from 35
breed varieties of Sri Lankan rice. It was reported that redrice breeds had stronger effects compared to white rice
breeds 39). Red rice’s inhibitory effects to form AGEs is
associated with cyanidine-3-O-glucoside (C3G 40). Additionally,
14 rice breeds including black rice, red rice, brown rice, andwhite rice are reported to have inhibitory effects on theformation of AGEs 41). Inhibitory effects on the formation of
AGEs by rice extract liquid are recognized in brown rice orwhite rice, which does not contain C3G. It is presumed thatthe effect is associated with polyphenol 41). FA is an ingredienteasy to ingest from grains which are a staple food. However,
it is possible that a large volume intake of grains mightcause excessive consumption of carbohydrates. It is necessary
to use refined FA or to be careful about the volume ofcarbohydrate contained in grains, in order to inhibit aging
and aggravation of diseases under the condition sufferingfrom strong glycative stress.

Protective effect on albuminous degeneration
caused by glycation
It has been reported that FA has an inhibitory effect todevelop neurotic diseases including epilepsy, depression,ischemia-reperfusion injury, Alzheimer’s disease (AD), andParkinson’s disease 42). AD is caused by glycative stress ormalfunction of mitochondria. Based on this, anti-oxidative
components are utilized to treat or mitigate AD. FA is one of
the anti-oxidative materials existing on the cell wall of plants
and prevents anti-inflammatory action and free radicals. FA
inhibits aggregation of amyloid β(Aβ)and has effects to keep
maintain protein structure 43). In addition, FA and its derivatives
inhibit the aggregation and fibrillization of Aβ 44). Furthermore,
FA is reported to pass blood-brain barrier 4 5). In this study,
it was clarified that FA inhibits a polymerization followedby protein cross-linking formation by glycated lysozyme,

while reinforcing the OPH activity effective in decomposing
glycated proteins. These results show a possibility that FA
prevents protein changing structures due to glycation,
resulting in protecting proteins. Individuals with diabetes
under a strong glycative stress are suffering from higher
rate of AD, compared to non-diabetes 46). Prevalence of
intracerebral AGEs of AD patients is three times higher
than in healthy subjects 47). Moreover, considering that those
whose accumulation of AGEs on skin is large in quantity
have a higher rate of mild cognitive impairment, and it
is presumed that an enhancement of glycative stress is
associated with development of AD 48). Based on this, there
is a possibility that protein protection by glycative FA isconcerned with preventing diabetics or aged people understrong glycative stress from developing neurotic diseases.  

Limitations of study
FA’s inhibitory effects of glycative reaction, inhibitory
effects of forming glycative protein cross-linking, as well as
enhancement effects of OPH activity validated in this study
are all in vitro test results. Further validation is necessary to
clarify the usefulness of FA administered continuously byhumans about the inhibitory effect of glycation.

Conclusion
FA has an inhibitory effect AGEs and glycation reaction
intermediates by the glycative reaction system with HSA,
collagen, and elastin as model proteins, protein crosslinkings, and enhancement OPH activity. There is a possibility
that usefulness of FA to inhibit development of diabetic
complications and other neurotic diseases is associated with
anti-diastatic effects.

Statement of conflict of interests
This study was supported by Social Welfare Corporation
Kitabakai. FA was provided by Tsuno Food Industrial
Corporation.

Acknowledgements
For this study, promotions support was given by Public
Interest Incorporated Foundation, Ishyoku-Dogen Research
Foundation (https://isyokudogen-fnd.jp/ )_ 191 _

Anti-glycative Effect of Ferulic Acid

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