62 Biol. Pharm. Bull. 39, 62–67 (2016) Vol. 39, No.

Regular Article

Cholesterol-Lowering Effect of Calcium Alginate in Rats

Yoko Idota,a Yumi Kogure,a Takako Kato,a Mana Ogawa,b Shoko Kobayashi,b
Chihaya Kakinuma,a Kentaro Yano,a Hiroshi Arakawa,a Chihiro Miyajima,c
Fumiyoshi Kasahara,c and Takuo Ogihara*,a
a
 Faculty of Pharmacy, Takasaki University of Health and Welfare; 60 Nakaorui-machi, Takasaki, Gunma 370–0033,
Japan: b Research Center for Food Safety, Graduate School of Agricultural and Life Science, The University of
Tokyo; 1–1–1 Yayoi, Bunkyo-ku, Tokyo 113–8657, Japan: and c Kimica Corporation; 2–4–1 Yaesu, Chuo-ku, Tokyo
104–0028, Japan.
Received June 23, 2015; accepted October 8, 2015

We examined whether calcium alginate (Ca-Alg) reduces blood cholesterol levels in rats fed a high-
cholesterol diet. First, we examined taurocholate adsorption in vitro by various types of sodium alginate (Na-
Alg). High molecular-weight, guluronic acid-rich Na-Alg showed the greatest adsorption of taurocholate, and
therefore the corresponding Ca-Alg was chosen for the in vivo study. Rats were fed a high-cholesterol diet or
a Ca-Alg-containing diet for 2 weeks. Body weight and diet intake were measured, and the general condition
of the animals was monitored during this period. After 14 d, the plasma concentration of cholesterol, portal
plasma concentration of bile acid, and bile acid in feces were measured. The plasma concentration of choles-
terol was significantly reduced in rats fed a 2% Ca-Alg-containing diet. Furthermore, the portal concentra-
tion of bile acid was significantly lowered in the 2% Ca-Alg group. A tendency for a Ca-Alg concentration-
dependent increase in fecal excretion of bile acid was also seen, although it was not statistically significant.
While several changes in biochemical parameters and histopathological findings were observed, all the values
remained within the physiological range. These results indicate that Ca-Alg is effective in reducing plasma
cholesterol. A possible mechanism would be enhanced fecal excretion of bile acid due to reduced intestinal
reabsorption, which in turn might stimulate bile acid synthesis from cholesterol in the liver, leading to a de-
crease in plasma cholesterol.
Key words alginate; cholesterol; bile acid

Heart disease and cerebrovascular disease account for about
one-quarter of all deaths in Japan,1) and a major factor in their
etiology is considered to be dyslipidemia.2–4) Indeed, dyslip-
idemia can be considered predominantly a modern lifestyle-
related disease.5,6) Therefore, there is considerable interest in
food additives or health foods that decrease cholesterol ab-
sorption or promote cholesterol excretion. Possible candidates
include alginate (Alg), a cationic intercellular polysaccharide
derived from brown seaweed.7,8)
We have compared the effects of sodium alginate (Na-Alg)
and calcium alginate (Ca-Alg) in promoting excretion and
decreasing absorption of Sr and cesium (Cs) in rats.9) Both
additives increased the excretion of Sr, though Cs concen-
tration was significantly reduced only in the Ca-Alg group.
We also measured the plasma concentration of cholesterol in
that study, and interestingly found that Ca-Alg has a greater
cholesterol-lowering effect than Na-Alg.9)
At present, Na-Alg is widely used as a food additive (thick-
ening agent) and as a health food to decrease blood cholesterol
and inhibit weight gain.10–12) However, sodium intake is asso-
ciated with hypertension,13) and Ca-Alg may be preferable to
Na-Alg, if it has an equivalent pharmaceutical effect. There-
fore, in this study we first examined various types of Na-Alg
to evaluate their bile acid-absorbing ability in vitro. Based on
the results, we selected high-molecular-weight, guluronic acid-
rich Ca-Alg for the in vivo study to examine its cholesterol-
lowering effect in rats given a high-cholesterol diet.
* To whom correspondence should be addressed.  e-mail: togihara@takasaki-u.ac.jp
MATERIALS AND METHODS
Chemicals and Animals Cholesterol, sodium cholate,
cholesterol E-test WAKO and total bile acid-test WAKO
were purchased from Wako Pure Chemical Industries, Ltd.
(Osaka, Japan). [G-3H]Taurocholic acid was purchased from
PerkinElmer, Inc. Life Sciences (MA, U.S.A.). High-molec-
ular-weight Ca-Alg rich in guluronic acid and various types
of Na-Alg, i.e., high-molecular-weight Na-Alg containing
guluronic acid and mannuronic acid (Na-Alg HMW-G+M),
high-molecular-weight Na-Alg rich in guluronic acid (Na-Alg
HMW-G), high-molecular-weight Na-Alg rich in mannuronic
acid (Na-Alg HMW-M), low-molecular-weight Na-Alg con-
taining guluronic acid and mannuronic acid (Na-Alg LMW-
G+M), low-molecular-weight Na-Alg rich in guluronic acid
(Na-Alg LMW-G), and low-molecular-weight Na-Alg rich
in mannuronic acid (Na-Alg LMW-M), were supplied by
Kimica Corporation (Tokyo, Japan). Colestimide, a cationic
polymer used to treat hypercholesterolemia, was purchased
from Mitsubishi Tanabe Pharma (Osaka, Japan). Normal rat
diet (CE-2) was supplied by Clea Japan Inc. (Tokyo, Japan).
All other chemicals and solvents were analytical-grade com-
mercial products.
The animal study was performed according to the Guide-
lines for the Care and Use of Laboratory Animals at Takasaki
University of Health and Welfare and approved by the Com-
mittee of Ethics of Animal Experimentation of the University.
Specific-pathogen-free male Wistar rats at six weeks of age
were purchased from SLC Japan (Hamamatsu, Japan), housed
2 animals per cage in a room kept under controlled conditions
© 2016 The Pharmaceutical Society of Japan
 Biol. Pharm. Bull.
Vol. 39, No. 1 (2016)63

(temperature of 23±3°C, humidity of 50±20%) with a 12-h Darmstadt, Germany) at 7500×g for 15 min. The amount of
light/dark cycle, and given free access to food and water. taurocholic acid in the filtrate was determined by radioactivity
In Vitro Experiments Solutions containing 2 n M [G-3H]- measurement using a liquid scintillation counter (LSC-6100;
taurocholic acid, 0.1 μM sodium taurocholate, and various Aloka, Wallingford, CT, U.S.A.). It should be noted that in
types of Na-Alg (3 mg) in 1600 µL phosphate-buffered saline vitro absorption assay of cholesterol was not carried out be-
were incubated at room temperature for 30 min, and then cause cholesterol did not pass through the filter (either because
passed through an Amicon Ultra 10k (Merck Millipore, of micelle formation or adsorption on the filter).
In Vivo Experiments One week after purchase, rats
Table 1. Comparison of the Adsorption Capacity of Different Types of weighing 113 to 152 g were randomized into 6 groups (n=6
Na-Alg, in Terms of Adsorption Rate of Taurocholic Acid each): normal diet group; high-cholesterol diet group (0.5%
(w/w) cholesterol, 0.25% (w/w) sodium cholate); three Ca-Alg-
Taurocholic acid adsorption rate containing diet groups (0.5% (w/w) cholesterol, 0.25% (w/w)
(% of control) sodium cholate, 0.5, 1 or 2% (w/w) Ca-Alg) and a colestimide-
containing diet group (0.5% (w/w) cholesterol, 0.25% (w/w)
Control 0.0±26.2
sodium cholate, 0.5% (w/w) colestimide). Rats received the
Low MW M+G 54.8±10.4
appropriate diet for two weeks. Body weight and diet intake
M-rich −0.1±48.1
were measured and the general condition of the animals was
G-rich 26.4±14.4
monitored during this period.
High MW M+G 78.7±1.7**
After 7 d and 14 d, 0.5 mL of blood was withdrawn from
M-rich 35.6±42.5
G-rich 82.1±3.5**
the jugular vein with a heparinized syringe under anesthesia
induced with diethyl ether, and centrifuged at 1700×g for
The data are mean±S.D. (n=3). The significance of differences from the control
was determined by means of Dunnett’s test. ** p<0.05. We used filters that had ad-
10 min to obtain plasma. After 14 d, 0.6 mL of blood was col-
sorbed excess amounts of taurocholate (cold) in advance. lected from the portal vein after laparotomy under anesthesia,

Fig. 1. (a) Time Courses of Cholesterol Concentration in Plasma during the Experimental Period, and (b) Differences between Groups at the End of
the Two-Week Period
Rats were fed normal diet, high-cholesterol (Cho) diet, or high-Cho diet containing Ca-Alg or colestimide. The data are mean±S.D. (n=6). The significance of differ-
ences from the high-Cho diet group was determined by means of Dunnett’s test. * p<0.05.
64 Biol. Pharm. Bull. Vol. 39, No. 1 (2016)

and the plasma concentration of cholesterol was measured.
Feces (n=2 rats/cage) were collected during the final 24 h of
the feeding period, dried in a lyophilizer, then pulverized and
heated in ethanol for two hours at 70°C. The heating process
was repeated four times, and bile acid in the pooled extract
was measured.
In accordance with the original Folch method,14) 2000 µL
of chloroform–n-methanol (2 : 1, v/v) was added to a 5 mL
tube containing 100 mg liver. Samples were homogenized in
a Polytron homogenizer (Kinematica, Lucerne, Switzerland),
and incubated overnight at 4°C. Four hundred microliters of
KCl 0.8% was added and the samples were vortexed and in-
cubated for 1 h at 4°C. Then, 500 µL of the lower phase was
evaporated under vacuum. The residue was taken up in 500 µL
chloroform–n-methanol (2 : 1, v/v), and a 100 µL aliquot of the
solution was evaporated under vacuum. The residue was taken
up in 100 µL of isopropanol, and the cholesterol concentration
was determined.

Table 2. Body Weight, Cumulative Amount of Diet Intake and Biochemical Parameters in Rat Plasma during Two-Week Feeding Period

Parameter Time (d) Normal High-Cho +Ca-Alg 0.5% +Ca-Alg 1% +Ca-Alg 2% +Colestimide 0.5%
Body weight (g) 0 133±13 132±13 132±13 132±10 132±11 131±6
7 175±14 174±13 169±12 174±7 172±12 170±5
14 202±15 196±13 196±13 199±4 199±16 195±6
Cumulative amout 0 0.0±0.0 0.0±0.0 0.0±0.0 0.0±0.0 0.0±0.0 0.0±0.0
of diet intake (g/ 7 107±3.9 104±2.8 99.8±1.8 106±1.7 102±8.6 96.1±2.8
body) 14 224±7.2 213±6.1 211±6.7 219±6.5 219±17 205±6.8
TG (mg/dL) 0 40.8±12.3 68.3±7.7 59.0±40.1 59.7±24.3 64.3±27.7 54.0±19.8
7 78.8±22.5 86.2±32.7 65.0±16.9 86.8±28.7 71.0±34.2 81.0±7.2
14 43.8±17.9 44.0±52.7 30.0±9.3 39.5±18.4 31.5±11.5 39.0±11.5
LDLC (mg/dL) 0 11.4±1.6 11.4±2.5 10.9±1.1 11.9±1.1 11.5±2.1 12.5±1.4
7 10.2±1.4 21.0±7.3 18.2±4.4 20.5±4.7 18.7±4.6 13.6±2.0
14 7.3±0.9 11.4±3.8 11.4±2.0 13.4±2.8 12.3±2.6 8.8±1.6
HDLC (mg/dL) 0 52.6±7.3 47.0±0.9 44.6±6.0 51.6±3.8 53.8±7.9 48.2±2.7
7 45.5±5.8 40.1±3.7 39.0±6.3 41.6±3.1 42.8±5.0 36.3±4.3
14 43.9±5.0 39.5±2.9 41.7±6.7 43.2±2.9 46.0±8.2 36.2±4.7
PL (mg/dL) 0 163.2±16.6 166.0±13.7 155.0±22.0 169.7±10.8 173.8±9.9 156.2±15.6
7 153.0±18.7 156.5±17.6 148.8±6.0 160.3±5.2 154.8±12.0 138.3±9.3*
14 134.7±14.7 130.8±22.0 129.8±11.6 141.5±5.2 139.7±9.7 117.5±10.0
NEFA (µEq/L) 0 0.45±0.09 0.49±0.10 0.45±0.11 0.46±0.19 0.47±0.15 0.45±0.12
7 0.55±0.05 0.66±0.15 0.64±0.20 0.64±0.09 0.66±0.11 0.57±0.04
14 0.73±0.26 0.56±0.18 0.58±0.17 0.65±0.16 0.55±0.18 0.62±0.23
Na (mmol/L) 0 141.0±2.3 141.0±3.1 141.3±2.8 141.4±3.0 142.2±1.9 141.4±4.9
7 138.2±1.9 139.8±1.5 138.4±1.3 138.6±1.2 139.4±1.8 138.7±2.2
14 142.1±1.1 142.9±1.1 144.4±7.1 144.0±4.4 141.8±1.0 142.2±1.5
K (mmol/L) 0 9.1±2.3 9.2±1.7 8.5±1.8 8.3±2.0 8.2±1.3 8.1±1.7
7 7.6±0.7 6.9±0.4 6.5±0.5 7.0±0.4 6.4±0.5 6.9±0.7
14 4.2±0.5 4.1±0.3 4.1±0.6 4.1±0.4 4.0±0.2 4.2±0.2
Ca (mmol/L) 0 9.5±1.0 7.8±2.3 8.6±0.7 9.2±1.0 10.3±1.1* 10.0±0.5*
7 10.2±0.2 11.1±0.1 10.8±0.3 10.6±0.2 10.9±0.2 10.5±0.7
14 8.7±1.7 10.4±0.7 10.2±0.5 10.7±0.6 10.6±0.2 10.2±0.5
Cl (mmol/L) 0 101.5±2.0 101.1±2.1 101.5±2.8 101.7±2.8 101.6±1.5 102.1±3.3
7 99.5±2.0 99.3±1.8 98.4±1.2 98.1±1.7 99.4±0.8 99.3±1.7
14 99.7±1.0 99.1±1.5 100.9±5.5 100.2±2.7 98.1±1.3 99.4±1.1
Cre (mg/dL) 0 0.19±0.02 0.17±0.06 0.20±0.01 0.20±0.01 0.19±0.01 0.19±0.01
7 0.21±0.01 0.23±0.01 0.21±0.01* 0.22±0.01 0.23±0.02 0.21±0.01
14 0.22±0.03 0.27±0.02 0.25±0.02 0.27±0.02 0.26±0.02 0.26±0.01
ALP (U/L) 0 2193±100 1903±617 2079±257 2135±209 2201±262 2195±130
7 2101±61 2745±228 2489±253 2613±104 2498±297 2533±234
14 1742±370 2185±244 2210±311 2129±204 2130±276 2286±190
GLU (mg/dL) 0 152.8±8.8 127.5±40.1 147.8±11.7 144.8±12.3 155.3±10.2 148.5±8.1
7 169.2±11.1 157.3±11.9 154.3±9.4 161.7±4.2 170.7±10.6 159.8±17.8
14 88.5±46.3 136.0±27.3 130.5±18.3 142.3±16.4 140.2±16.3 105.2±14.0*
ALT (U/L) 0 42.5±6.7 32.3±11.1 37.8±6.0 38.7±3.7 43.0±6.2 38.7±6.6
7 42.5±4.5 64.7±45.0 53.2±9.4 55.5±16.2 62.2±18.0 39.3±8.4
14 40.7±10.6 46.2±9.5 45.5±8.4 49.2±6.9 50.0±6.5 44.7±10.2
AST (U/L) 0 75.8±6.0 66.7±19.7 82.2±6.1* 73.7±4.1 74.0±3.4 77.8±5.8
7 75.7±6.0 117.8±80.4 103.7±30.6 94.3±18.2 97.8±19.5 74.3±6.9
14 71.5±16.3 72.8±10.9 77.3±13.3 76.8±7.0 73.3±5.9 75.2±3.5
The data are mean±S.D. (n=6). The significance of differences between the high-Cho group and the Ca-Alg containing groups or the colestimide group was determined by
means of Dunnett’s test. * p<0.05.
 Biol. Pharm. Bull.
Vol. 39, No. 1 (2016)65
Fig. 2. (a) Amount of Bile Acid in Feces of Rats after Two-Week Feed-
ing Period with High-Cho Diet or High-Cho Diet Containing Ca-Alg,
(b) Portal Plasma Concentration of Bile Acid in Rats after Two-Week
Feeding Period with High-Cho Diet or High-Cho Diet Containing Ca-Alg
(a) The data are mean±S.D. (n=3). Differences were not statistically significant.
(b) The data represents mean±S.D. (n=4–6). The significance of differences be-
tween the high-Cho diet group and the groups fed high-Cho diet containing Ca-Alg
was determined by means of Williams’ test. * p<0.05.
For safety monitoring, blood samples withdrawn after 14 d
were used for the measurement of biochemical parameters,
namely aspartate aminotransferase (AST), alanine amino-
transferase (ALT), total cholesterol (Cho), triglyceride (TG),
phospholipid (PL), non-esterified fatty acid (NEFA), high den-
sity lipoprotein (HDL), low density lipoprotein (LDL), blood
glucose (GLU), Ca, Na, potassium (K), and chloride (Cl).
After blood sampling, the animals were euthanized by ex-
sanguination (n=3 each). Heart, liver, lung, kidneys, spleen,
submandibular glands, parotid glands, mesenteric lymph
nodes, stomach, intestines, testes, epididymis, thyroid glands,
adrenal glands, white adipose, tissue and brown adipose tissue
were removed and immediately fixed in 10% neutral buffered
formalin. The tissues were immersed in paraffin, and cut at
4 µm thickness. The slices were stained with hematoxylin and
eosin, and examined microscopically.
Analytical Methods Biochemical parameters were mea-
sured with commercial kits using an automated analyzer
(BioMajesty™ JCA-MB6050, JEOL Ltd., Tokyo, Japan) and
blood testing apparatus (XT-2000iV, Sysmex Corporation,
Kobe, Japan). DeterminerL TGII BMS was purchased from
Kyowa Medex Co., Ltd. (Tokyo, Japan). Oculusauto CRE was
purchased from Shino-Test Corporation (Tokyo, Japan). Anal-
ysis kits for other biochemical parameters were purchased
from Wako Pure Chemical Industries, Ltd.
Statistical Analysis Data are expressed as the
mean±standard deviation (S.D.). Statistical comparisons were
made using the Dunnet test and the Williams test. Values of
p<0.05 were considered significant.
RESULTS
In Vitro Experiments The amount of bile acid adsorbed
by high-molecular-weight Na-Alg was greater than the amount
adsorbed by low-molecular-weight Na-Alg. The recovery of
bile acid in the filtrate was least (less than 20% of the control)
in the case of Na-Alg HMW-G (Table 1). We therefore se-
lected Ca-Alg HMW-G for the in vivo experiment.
Effect of Calcium Alginate (Ca-Alg HMW-G) on
Parameters of Lipid Metabolism in Rats
Plasma Concentration of Cholesterol in Rats
Plasma concentration of cholesterol was increased at one
week in all groups except the control group (fed a normal
diet). In the group fed colestimide-containing diet, the extent
of the increase was less than in the others. Between one and
two weeks, the plasma concentration of cholesterol declined in
all groups, and the group fed the Ca-Alg 2% diet showed the
greatest decrease during this period (Fig. 1a). In the groups
fed diets containing Ca-Alg, the increase of cholesterol con-
centration over the 2-week period appeared to be inversely
dependent on the concentration of Ca-Alg in the diet. In the
groups fed Ca-Alg 2% diet and colestimide-containing diet,
the plasma concentrations of cholesterol at 2 weeks were
128.5±14.8% and 131.4±19.6% of the initial value, respec-
tively, and were significantly lower than that in the group fed
high-cholesterol diet alone (161.3±24.0% of initial value) (Fig.
1b).
Plasma Concentration of Other Biochemical Parameters in
Rats
After one week, PL concentration in plasma of the
colestimide-containing diet group was significantly lower than
that of the high-cholesterol diet group, but no difference was
observed at two weeks. No marked difference in any other
biochemical parameters related to lipid metabolism was ob-
served among the groups (Table 2).
Excretion of Bile Acid in Feces and Portal Plasma
Concentration of Bile Acid in Rats
Bile acid excretion in feces tended to increase depend-
ing on the concentration of Ca-Alg in diet (high-cholesterol
diet alone, 194.8±33.6; Ca-Alg 0.5%, 190.8±26.2; Ca-Alg
1%, 212.1±5.8; Ca-Alg 2%, 262.2±82.2 µmol/d/2 rats/cage),
but the differences were not statistically significant (Fig.
2a). In the group fed the 2% Ca-Alg diet, the portal plasma
concentration of bile acid was significantly decreased to
203.4±97.0 µmol/L, compared to the high-cholesterol diet
group (298.7±77.8 µmol/L) although the decrease did not ap-
pear to be strictly dose-dependent (Fig. 2b).
66 Biol. Pharm. Bull. Vol. 39, No. 1 (2016)

Fig. 3. Liver Weight (a) and Total Cho in Liver (b) in Rats Fed the High-Cho Diet or the High-Cho Diet Containing Ca-Alg for Two Weeks
The data are mean±S.D. (n=6).

Fig. 4. Representative Histochemistry of Periportal Hepatocytes in Rats Fed High-Cho Diet for Two Weeks
Panel (b) is an enlargement of panel (a). Microvesicular steatosis was increased in the high-Cho diet group, but remained within the normal physiological range.

Table 3. Histological Findings in Rat Liver and Kidney after Two-Week Feeding Period

Ca-Alg Colestimide
Normal High-Cho
0.5% 1% 2% 0.5%
Histological findings 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3
Liver
Microvesicular steatosis in — — — ± ± ± — ± ± ± ± ± — ± ± — ± —
periportal hepatocyte
Kidney
Basophilic renal tubules, focal — — — — — — — — — ± — — — — — ± — —
Angiectasis — — — — — — — — — — ± — — — — — — —
Deposition of hyaline drop — — — — — — — — — ± ± — ± — — — — —
Grade of changes: —; no, —/±; within normal range, ±; slight. Cho: cholesterol.

Liver Weight Gain and Total Cholesterol in Liver
There was no difference in liver weight gain (high-choles-
terol diet alone, 9.8±1.0; Ca-Alg 2%, 10.0±1.1 g) or total cho-
lesterol in liver (high-cholesterol diet alone, 1437±432; Ca-Alg
2%, 1629±407 mg) between the groups fed high-cholesterol
diet and Ca-Alg 2% diet (Fig. 3).
Safety Assessment There were no significant differences
in weight gain or diet intake among the groups during the
2-week experimental period (Table 2). No change in general
physiological condition was observed in any animal in any of
the groups. After two weeks, GLU concentration in plasma of
the colestimide-containing diet group was significantly lower
than that of the high-cholesterol diet group. Several differ-
ences, including in initial values, were observed among other
parameters, but all values were within normal physiological
ranges (Table 2). Microvesicular steatosis was increased in
the high-cholesterol diet group and in the Ca-Alg containing
groups, but remained within the physiological range (Table 3,
Fig. 4).
DISCUSSION
There are several kinds of Alg, namely low- and high-mo-
lecular-weight types containing mainly β-D -mannuronic acid
 Biol. Pharm. Bull.
Vol. 39, No. 1 (2016)67
(M-rich), mainly α-L-guluronic acid (G-rich) or both (M+G).8)
In order to identify the best Ca-Alg for the in vivo experiment,
we first carried out an in vitro study to compare the tauro-
cholate absorption capacity of these types. For this, we used
Na-Alg, because Ca-Alg is virtually insoluble in water. Tau-
rocholate was selected based on preliminary experiments with
colestimide, because cholesterol failed to pass through the
filter. Based on the results, we chose high-molecular-weight
G-rich Ca-Alg, which is commercially available, for the in
vivo experiments.
When 2% Ca-Alg diet was given to rats for two weeks, the
portal plasma concentration of bile acid was significantly low-
ered; further, the amount of bile acid in feces was increased
in all Ca-Alg groups (though without statistical significance).
Therefore, Ca-Alg appears to promote excretion of bile acid
in feces, presumably by binding to the bile acid and inhibiting
its intestinal reabsorption. Moreover, the increase of plasma
cholesterol level at 2 weeks was significantly reduced in
the 2% Ca-Alg group, to a level similar to that in the 0.5%
colestimide group. We speculate that reduced reabsorption of
bile acid caused a decrease of bile acid concentration in the
portal vein, which in turn would induce increased bile acid
synthesis from cholesterol in the liver, with a concomitant re-
duction of cholesterol concentration in plasma.
It is well known that 95% of bile acid eliminated via the
bile duct to the intestine is reabsorbed into the body,15,16) and
gastric–hepatic circulation is the main supply source to main-
tain bile acid homeostasis in the body. Therefore, inhibition
of reabsorption of bile acid by Ca-Alg may be an efficient
mechanism for the reduction of plasma cholesterol.
We found no significant differences in weight gain, diet in-
take, or biological parameters among groups. Moreover, there
was no change in total amount of cholesterol in the liver or
liver weight gain. While several changes in biochemical pa-
rameters and histopathological findings were observed, all the
values remained within the physiological ranges. Furthermore,
the animals in all groups showed no apparent abnormalities.
These results indicate that Ca-Alg is safe when taken daily for
2 weeks at the 2% concentration level. We could not confirm
what types of lipoproteins were reduced by Ca-Alg treatment,
because LDLC and HDLC fluctuated during the test period.
Further study would be needed to resolve this issue.
In conclusion, our results indicate that 2% Ca-Alg in the
diet reduces the increase of plasma cholesterol in high-choles-
terol-fed rats. A possible mechanism would be enhanced fecal
excretion of bile acid due to reduced intestinal reabsorption,
which in turn might stimulate bile acid synthesis from choles-
terol in the liver, leading to a decrease in plasma cholesterol.
In this study, 2% Ca-Alg in the diet was found to be effective.
Ca-Alg is convenient to take, because it is effective in solid
form, unlike Na-Alg. Furthermore, Ca-Alg has been con-
firmed as safe for use as a food additive, and there is no risk
of hypertension due to increased sodium intake. Therefore,
Ca-Alg is expected to be suitable as an additive or functional
food for long-term use.
Acknowledgment This work was supported by
JSPS Grant-in-Aid for Challenging Exploratory Research
(KAKENHI) Grant Number 25560062.
Conflict of Interest Chihiro Miyajima and Fumiyoshi
Kasahara are employees of Kimica Corporation. The others
have no conflict of interest.
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