Title Effect of Essential Fatty Acids and Pridoxine on the Formationof Gallstones, Especially Cholesterol Stones

Author(s) MARUYAMA, IZUMI

Citation 日本外科宝函 (1965), 34(1): 19-34

Issue Date 1965-01-01

URL http://hdl.handle.net/2433/206454

Right

Type Departmental Bulletin Paper

Textversion publisher

Kyoto University

Effect of Essential Fatty Acids and Pyridoxine on the

Formation of Gallstones, Especially Cholesterol Stones

by

IχUi¥11 MARUY州主

From the 2nd Surgical Division, K、t山， Uni河川勺 MedicalSchool (Director：ドrりfDr. CHU)! KIMURA)

Received for Publication Nov. 10, 1964

I INTRODUCTION

19

The fundamental studies on the mechanisms of gallstone formation were established

by NAU！叫N and AscHOFF. In general, it is believed that local factors such as biliary

infections, bile stasis, damage to the gallbladder wall and so on lead to the decrease of

cholesterol holding power of gallbladder bile so that cholesterol precipitation may occur and

stones begin to form. However, cholecystitis without stones or cholelithiasis without biliary

infection are frequentlv encountered clinically. Many other investigators have tried to

elucidate the initiating factors of gallstone formation with disturbances of metabolism,

dysfunctions of the endocrine system, dysharmonies of the autonomic nピr、りU只 systemand

abnormalities of constitution. But the etiology of gallstone yet remains obscure.

In our laboratory, HIKASA et al.10l 1•l 16l 3n investigated the specific nutritional effects

of essential fatty acids (EFA) and recentlv have reached to the conclusion that cholesterol

esterified with EFA especially with tetraenoic acid, i.e. metalコolicallyactive cholesterol, i只

the direct precursor of adrenocortical hormones. On the other hand, bile acid is a main

end-product of cholesterol2> 3> 5> 49> 見け thatit is reasonable to suppose that cholesterol should

be metabolically active, i.e. it should be esterified with tetraenoic acid, for the biosynthesis

of bile acid. According to this supposition, HIKASA 13> has first surmised in 1960 that

gallstone may be due to deficiency or metabolic disturbances in EFA.. Deficiency and/or

metabolic disturbances in EF A lead to decreased biosynthesis of bile acid. Lecithin, which

contains EFA as its component, may be decreased in these stat引 ． 点け that cholesterol

precipitation may occur and stones begin to form.

The present study was designed to clarify the influences of EFA upon the gallstone

formation.

II MATERIALS AND METHODS

MATERIALS

1) Clinical Experiment

Gallbladder biles obtained aseptically during surgery were analysed. The patient只

were composed of 39 cases of gallstones and as a control group 21 cases of peptic ulcer,

admitted at the Second Surgical Division, Kyoto l fniversity Hospital. Thev were all without

distinct disturbances of liver functions. Further, the hepatic biles from the common bile

duct through the draining tube were analysed.

20 日f.:（，；トFドi;:r'J'.i :'j'¥3 t誉市1号

2) Animal Experiment

Male albino rats of Wistar strain supplied by the Animal t冶nterin Kyoto l Tniversity

were housed in single, screen-bottom cages at 20° C. They were fed a rat chow until

their body weight reached about 40 to 50 mg, then they were divided randomly into three

groups EFA diet, EFA-deficient diet and pyridoxine-def icient EF A diet groups (5 to 9

animals in each group). The composition of each diet is listed in Table 1, 2 and 3. As

the source of EFA. a purified and peroxide free sesame oil was used. The sesame oil

contained 48.9 % linoleic acid, 0. 7 % linolenic acid, and no other polyunsaturated fatty

acids.

Table 1 The Composition of the Diet

Starch

Vitamin-free C九%引nSesame (Ji[

:-:alt Mi,t11rぞ

Vitamin Mixture < [1.,[ine ( ＼，］，』rul1

火 lヘIid＜ 代in<frf'f'

EFλDiet

60°0

16

20

3 o.:;

o.s

Table 2 Vitamin :V11リ11re(in I g)

Vitamin . .¥

!31

82

l3s

B12

D

E

K

Niacin

Pantothenic入ιirl

Foli】C .¥, irl

Table 3

¥.";i(J

¥.";iHJ'< J,

K.HF<>,

（、lll ,r J'( >,,・HJ J

Cd. Lactate M店、（），

2500 I.U.

1.0mg

l.5mg

!.Omg

1.0;

37.5mg

200 I.U.

!.Omg

0.2mg

10.0mg

2.5田g

0.5mg

S;dt Mi,t11n’（in 1000 g)

46.3gm

92.0

253.0

143.0

369.0

70.4

EF A-deficient Diet Iヘridoxine-deficientEL'¥. Diet

80°0 60%

16 16 。 20

3 3

0.5 0.5権

0.5 0.5

All rats wen' thus maintained for about

3 months. During the course of feeding

period, EFA-deficient and pyridoxine-

deficient EFA diet groups exhibited the

characteristic signs of EFA deficiency and

pyridoxine-deficiency. At the end of this

period, all rats were fastened for 12 hours

and sacrifized by bleeding from the aorta.

The li¥-er slices were excised and weighed

immediately by microbalance. The bio-

chemical analysis were made with these

liver slices.

¥IETHODS

1) Extraction and Determination of Bile

人ciclin Bile

Bile acids were extracted by a modi-

fication of the procedure reported by Mos-

H,¥cH・iaJ. Twenty volumes of 99% ethanol

was added to the bile (or homogenated

liver slices), and the mixture was heated Kl ~6.:J

on a steam bath for 1 hour. The mixture

was filtered and the fl叫iduewas washed twice with 5 ml of petroleum ether (bp. 30° to

50 C) Io remove neutral lipids・ Thissolution w川 thenacidified with 3N-HCI and further

EFFECT りFEF礼 （）火 THE Iごりl{M.¥Tlり』＼ < >F 1; ¥I.I Jずl<>!¥I・メ 21

extracted with petroleum ether to remove fatty acids. The acidic alcohol ph11se was then

evaporated on the steam bath under a current of air. The residue was dissolved in 5 %

NaOH and hydrolysed at 120°仁 for3 hours. The hydrolysate was acidified with 3N-

HCI, and extracted 4 times with 10 ml of ethyl ether. The ether extract was washed

with distilled water to rem川 ＇l' chloride and dried over anhydrous sodium sulfate. A suita-

ble aliquot of this extract was pipetted into a test tube, and evaporated on the吋引 rnbath.

FivP ml of 65% H，メ（)1 were added and after heating 60 ± 1 C for 15 min., the optical density was read at 3200 and 3850 ...-¥ with the Br·川、 ~ L \ :--i DU quartz spectrophotometer.

2) Extraction of Lipids and Determination of Other Lipids in Bile

a) The extraction of lipids is illustrated in Fi巴.1.

b) Fractionation of Bile Lipids with Silicic Acid Column Chromatography

Fractionation of bile lipids were performed by a modification of HIRSCH and A.HRENS

method 19>. Petroleum ether solution containing 1 % (80 ml), 10 % (100 ml) and 20 ・；シ

(150 ml), were employed to elute successi¥・elv cholesterol 引 Tピf'-, triglycerides and free

cholesterol. Phospholipids (lecithin) were subsequently eluted with 300 ml methanol.

Fractionated bile lipids were then saponified with 10% KOH-ethanol at (;r1ぐ for1

Fig. 1 E川町ktinn of Bile Lipid、

Fraction a le 、、1d1silicic acid column chn 1111at＜＇日raph、

Bile (2 ml)

}frat at 6:i C for 5 min. in 40 ml ！ ~I山r＼田luti口n

Filtrate

I Extract w出 20ml petrole…い times

I Petroleum Ether おl11tJ<>llj •Li 1叫 ｝

ト古江戸川1f,・ with 10°; KOH-ethanol at 60 C for J hour

i Extract川 ith 20 ml petn

I 3 ti

Residue Petroleum Ether 片山ill• 川〔TotalChoJe,terけ11

¥cu.l1f1・ w】th3×－HCI

Extract 川thIO ml petroleum ether

3 times

Wash "ith distilled日・atert" ice

λ11'orpti"n of the 山 il目、、ith

sodium sulfate

22 日本外科宝函第34巻第1号

hour. After extraction of nonsaponifiable materials (esterified and free cholesterol) from

the alkaline solution with petroleum ether, fatty acids contained in cholesterol esters, glyc-

erides and phospholipids were removed by acidification with 3N-HC! and subsequent ex-

traction with petroleum ether, respectively. The extracts were washed with distilled water

and dried over anhvdrous sodium sulfate.

c) Determination of Cholesterol

Esterified, free and total cholesterol were determined by KINGSLEY’s method25> of

LIEBERMANN-BURCHARD reaction.

d) Determination of Fatty Acids

Fatty acids from cholesterol esters, glycerides and phospholipids, and total fatty acids

were weighed by microbalance, after the addition of hydroquinone and evaporation to

dryness under nitrogen.

3) The Gas-liquid Chromatographic Analysis of Fatty Acids

The extracted fatty acids were methylated with freshly prepared 2 ＇~＞， H2SO,-methanol

at 70''C for 90 min .. The methyl esters of the fatty acids were dissolved into petroleum

ether and chromatographed in a SH!MADZU Seisakusho Model GC-1-B instrument equipped

with a hydrogen frame ionization detector. A 150 cm×6 mm i. d. column packed with

25% diethylene glycol succinate polyester coated on Shimarite (60 to 80 mesh) was used.

The flash heater was maintained at 300' C, the column at 210°C, and the detector at

240°C; the nitrogen flow rate was 30 ml/min.×2, the gas pressure 3 Kg;cm2. The fatty

acids from phospholipids and the total fatty acid were analysed by gas-liquid chromato-

graphy. Each component of the fatty acids separated by gas-liquid chromatography was

identified with the standard samples offered from National Institute of Health and Hormel

Institute in l T S.λ.. The details of this procedure were reported previously15>.

4) The Analysis of Gallstones

The analysis of gallstones were performed by a modification of the method reported

by NrsHIMlH<.-¥ 39>. The 伊 llstonescontaining mげ 70% of cholesterol were grouped into

cholesterol stones.

III RESULTS

1) Clinical Experiments

Gallbladder biles were tested microbiologically, and the patients were divided into the

groups of with or without biliary infection. The results of the experiments are listed in

Table 4.

a) Cholesterol

主｜】りut98':>o of cholesterol in bile was fr町 andthe esterified cholesterol was detected

only traces. Total cholesterol in bile did not significantly increase statistically in cholesterol

stones when biliary infection was absent, while it decreased significantly in the presence

of biliary infection.

b) Bile Acid

Total bile acid decreased moderately in the bile with cholesterol stones when without

biliary infection, while it did strikinglv with biliary infection. Moreover, change in com-

position of bile acids, that is the ratio of dihydroxycholanic acid to trihydroxycholanic acid

FドドFt、T(Jド EF人 ＜J>; THE FORMATf< J:--; < !F r;.¥LLSTONES 23

Table 4 Bicchemical ¥n.1k川、ofHuman Gallbladder Bile

ぐh()Jf'－；［ぞnolメt<lfltヘ ι1い‘》1.. 、te晶 r<》｜円l‘’( ・, 111tr《，l 、、1th ： 、、1th，’ll t 1 without

Biliary Infれ ti<111 Hili;irv lnft・cti"n ' Hil1;in・ !11fect1<>11

Ne』りiPatients

Tc山 iぐhc1lt'、tl'n,J mg/di

Total Bile .¥ml mg/ml I

flih、cln山、 .＼ncl'Trihydroxy., . .¥cid

Bile λι1d/Chc1le：、knol

Total Fatt、..¥c・id mg/ml

21 I 20

608士36 681士85

67.8士3.1 cl7.9土 1.0

1.18士0.08 1.39士0.11

11.68士2.06 7.75土1.67

24.39士2.01 II.I 1士2.18

＼’umhers after土 arestandard em'"・

6

261土63

ヨ］.L士6.0

0.90土0.10

10.20土3.04

1.94士1.90

6

671土181

42.8土6.3

1.10士0.09

9.67土2.51

19.57士l.62

(Di Tri), was not so different from that of control group in the cases of gallstones

without biliary infection, on the contrary, it decreased markedly when with biliary infec-

tion. Thus change in bile acids in the cases of cholesterol stones with biliary infection

is clearly different from that of without biliary infection.

c) The Ratio of Bile Acid to Cholesterol (B C)

As shown in Table 4, the ratio of R C was decreased siεnificantly in the bile with

cholesterol stones when without biliary infection. The ratio was less than 8. This value

is important because M1 Y ,¥KE"l has reported that the ratio of B C more than 8 was neces-sary to keep cholesterol in solution.

d) Fatty Acids

When lipid composition of bile was analysed by silicic acid column chromatography,

it was found that about 95 to 99 % of total fatty acid was contained in lecithin. Therefore,

quantitative change in total fatty acid was parallel with that in lecithin. Further, changes

in total fatty acid composition were almost identical to those of lecithin composing fatty

acids, as shown in Table 5. Total fatty acid was decreased in the bile with cholesterol stones in cases of without

biliary infection. When accompanied with biliary infection, it decreased more strikingly.

However, fatty acid composition was not so different fron】 thatof control group in either

Table 5 Composition of Total Fattv . .¥rnl and Fatt、主山l（＇，廿1川 1ll'rlin L町 ithinI%)

Patient Diag11<≫1、 c c c c c c (' c c c c F.itt、、 λι1d14・016・016 : I 18 0 18・118 : 2 18 : 3 20 : 3 20 : 4 22 : 0 22 : 6

一 一 一一一一一一一一 一一 一一 一

31.5 .t.2 6.1 20.0 23.3 0.3 2.1 5.-1 1.5 5.0 C間 II加 Ulcer

Lecithin 0.4 30.5 』.-1 8.:C 20.8 22.2 0.3 1.9 ー 勺 I. I 1.5 " -一一一一一

I i Total 1.3 38.8 5.3 6.3 13.5 4ドヲ ・〉 。.5 1.0 1.1 2.3 3.8 C川ぞ 2Peptic Ulcer

I Lecithin 0.9 38.2 4.9 1.0 15.2 21.9 0.2 1.1 4.6 1.8 3.9

I Cholesterol Total 0.5 36.0 ,).;:, 6.0 11.2 22.5 0.1 1.0 L3 2.5 6.3 C‘l吋；i 討ton~ with口ut

Infecti円n Lecithin 0.5 36.8 5.8 6.5 13.2 ~2 .0 0.5 I. I I.I ~.8 6.2 日一一一一一一一一一

-11. I C＂叩 4Stone withe nil 1

4.3 3.5 11.4 26.8 0.4 0.2 4.6 1.9 4.2

Infection Lecithin 0.2 40.8 3.8 7.7 12.l 23.6 0.3 0.4 1.6 4.7

日木外科宝函ヰiSJF会第1号

山崎（Tahle6). These results were agreed with the data reported by BLOMSTRAN04>,

although 蛇，・era!investigators have reported an increase of oleic and a decrease of linoleic

acid40＞川. Anvwav, the present results show that lecithin does not change qualitatively,

hut does quantitatively.

In a word, in the bile of patients with gallstones, especially with cholesterol stones

and without biliarv infection, decreased bile acid and lecithin were ohserved. When ac-

companied with biliary infection, however, decrease of bile acid and lecithin became more

strikinεand qualitative change in the bile acid was observed. Therefore, when hiliary

infection was not present, the composition of the bile with cholesterol stones was different

from when it was with hiliary infection. The changes in composition of the bile with

cholesterol stones may not be caused hy the infection.

2) Animal Experiment

It has already been demonstrated in our lahoratory that cholesterol esterified with EF A

especially with tetraenoic acid was a precursor of adrenocortical hormones10> 14> 16> 37>. Ac-

cording to this fact, it was surmised that deficiency in EFA. resulted the reduced synthesis

of bile acid from cholesterol in liver. Based on this presumption, the determination was

done on bile acid in the liver slices of the rats fed various diets.

日） Resting State

：＼~ compared with EF A diet只roup,marked decrease in total hile acid and elevated

ratio of I )i ・Tri were observed in EF A deficient diet group. In pyridoxine-deficient EF A

diet grou1】， however,patterns of bile acid were almost日imilarto those of EF A diet group

(Tahle 7).

h) Administration of ACTH Z

After the daily intramuscular administration of ACTH-Z 3 J.IJ. for consecutive 4

days, total bile acid decreased markedly in pyridoxine-deficient EFA diet group. On the

other hand, it decreased slightly in EF . .¥ diet group (Fig. 2).

Table 6 F.itt、.＼cid日 ・mp＜川ti川 i of Human Gallbladder Bile (%)

Control

¥o, of Patients 13

（、 14: 0 0.3士0.1

（‘ 16 ' 0 41.0士2.9（、 16:I 3.9土0.5( 18 0 '.?.8土0.6（、 18. I I 1.3士l.I （‘ 18・2 '.2'.?.I士l.2（‘ 18 3 0.5土0.2（、20 3 0.7士0.2( 20. 1 6.6士0.7（、22: 0 3.4士O目5（、22:6 4.7士0.6

河川.lwr'after士 arestandard <Tr t』，.，

('hi.It・、ternlStoneo without

Btl1.1r、Infection

15

0.5士0.1

31.8士3.0

I l土0.5

3.7士0.7

15.9士lI

2 l. 1士LO

0.1士0.1

1.2士0.2

6.2士0.5

2.Y土0.4

4.7士OA

Cho lesterりISt11nいwith

Bilian・ Intect1on

6

0.；）士0.'.2

3'.2.1士1.1

3.5士0.6

:i.9士0.6

17.0土1.1

22.I士1.0

0.4士0.1

'.?.3士0.5

7.8土0.6

:2.9二l::0.4

4.9土0.5

Pigment St《＞llC、without

Biliarv [nfectwn

1.6土0.I

5目』士0.9

17.2士0.I

:27.3士2.7

0.6±0.2

0.7土0.7

8.6土1.3

2.8士0.3

5.8ト0.7

EドドFぐTOF EFλUN THE FりR孔fλTI<>i¥ < 1F <;Auぷ［＇（） i¥'ES 25

EFえ Diet

Table 7 Bile A.cicl in the Liver ,,f Rats Fed Various Diets

EF A-deficient I Jwt lヘricloxine-cleficientEFA Diet

下山）． of Animals

Total Bile .¥cir! (w／、、%）

fl1h1 dn ＇＂ λじl《lTnh、dn山、.＼ml

η乙ーハリハU

m士ハ同i

d

ハUnU

内〆I

“

Qυ ハHv

ハHu

印

土戸、υ

ηペυ

－

nり 0.146土0.041

N’umbersι1fter土 arestandard errors.

0.26士0.04

N " w " E~孔 lliet

。IS

11111

。11・,

仁二二二二コ~

~

Pvridoxine IJef1cient EFA Diet EE~Defl口ent D』et

ACTH Z

－一一一一→:Jill/day for 1 days

Fig. 2 （、h.ingesin the Bile λじidLevels in the Liver of R‘iト FeelVa.rious Diets after the .¥clrni1ll';tr;it1けnof .＼（、TH-Z

6

0.45士0.04 0.23土0.05

According to these results, it is evi-

dent that EFA are necessary for the syn-

thesis of bile acid from cholesterol and that

pyridoxine deficiency accompanied by the

stress may affect this proc明丸

3) Clinical Application

Patients with gallstones were selected

for this experiment after surgical drainage

was done from the common bile duct. They

were not suffering from any distinct disturb-

ances of liver functions. They were fed

a basal diet (protein: 7 4.3 g, carbohydrate :

470.ll g, fat 10.0ε，2267 Cal.) after 7th

day of operation. Case 1 was administer-

ed both吋 valecithin 20 g and pyridoxine

hydrochloride 50 mg per day. Case 2 was

administered sC1y；ιl lecithin 20 g per day and

the other 2 patients were not administered.

Twenty grams of si"¥ya lecithin contained

8.52 g of total fatty acid: 2.004 g of palmitic acid, 0.444 g of stearic acid, ll訓 lI g of oleic

acid, U68 g of linoleic acid, and 0.460 g of Iinolenic acid according to同日ーliquidchromato-

graphic analv只is. Hepatic bile of these patient弓 werecollected forヒ1じry21 hours and

analysed.

Case 1 (treated with both同 valecithin and pyridoxine) : bile acid increased gradually

while cholesterol decreased, so that the ratio of B C was elevated (Fig. 3 a) .

Case 2 (treated with sδya lecithin only) : although both bile acid and cholesterol

increased, the increasing rate was higher in bile acid than in cholesterol，同 thatthe ratio

of B;C was elevated (Fig. 3 b).

Case 3 (non-treated) : change in bile acid level was slight and cholesterol was increased.

The ratio diminished gradually (Fig. 4 a).

（、ase4 (non-treated) bile acid was decreased markedly and the ratio diminished

(Fig. 4 b).

The ratio of B/C was elevated in all patients treated with EFA and pyridoxine, on

the contrary, in non treated patients the ratio of B （＿、 wasdecreased gradually. Thus EFA

and pyridoxine are effective for the treatm.ent of gallstones after surgery and have a

~1 2

" ~ 10 2 ぷu

""- ，；・司

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旦ッ

国

－ ~ 12 ω

“ ~ 10

－ 主ド'-' '¥. 6 ’U

足4

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雫ヨu 〈

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Lecithin － Pyndo x・ne- 2.1’ E ＼

"' E 15

τ；3 'u I 0 〈

ω ー『国 U.5

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~ II 13 15 17 days

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7 9 11 13 lo days

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7 9 11 13 15 17 19 l 1 days

Fi耳 3I b1 （川e2

3 5 7 9 II 13 15 17 days

Fig. 3 1a1 C‘t吋、 l

The !nflu!'ncc、pfEf. . .¥ nnd p、Iido日neupのn the C川 np<>,iti《川。fH11111:111 Hqx.11<・ Bile Fig. 3 and (

≪> <N

EFFECT OF EF,-¥ Uメ THEF<m:VIλTION OF GALL-iTt J:¥I・：メ 27

protective effect for the gallstone formation.

IV GAS-LIQUID CHROMATOGRAPHY OF BILE ACIDS

In the present chapter, separation of bile acids from human bile with the aid of gas司

liquid chromatography has been reported.

Method. Bile acids, extracted by the method described above, are methylated with

freshly prepared diazomethane. Then trifluoroacetates were prepared by heating the methyl-

esters in 0.1 ml of trifluoroacetic anhydride and 0.1 ml of pyridine in a glass stoppered

test tube at 30°C for 15 min.20>50>. The reagent was evaporated under nitrogen and ex-

tracted with ethyl ether. The ether solution was then washed with dil HCl and then

distilled water and evaporated to dryness. The residue was dissolved into acetone and

chromatographed in a SHIMADZU Seisakusho Model （正 1B instrument equipped羽 itha

hydrogen frame ionization detector. A 225 cm×4 mm i. d. column packed with 0.7ルnitrite silicone coated on Chromosorbも"111 (60 to 80 mesh) was used9>. The flash heater

maintained at 240仁， the column at 230 C, and the detector at 240°仁、 the nitrogen

flow rate was 30 ml/min.×2, the gas pressure 3 Kg/ cm 2.

Trihydroxy compounds gave 日omewhatlower responses with the detector, therefore,

it was necessary to use correction factors determined with a testmixture of known com-

position.

Results. Fig. 5 shows the separation of a testmixture. Although lithocholic acid has

almost the same retention time as deoxycholic acid, a good separation is obtained between

deoxycholic, chenodeoxycholic and cholic acid. Fig. 6, 7 and 8 show gas-liquid chromato-

grams from various patients. A~ illustrated in Table 8, bile acid composition of gallbladder

bile is not so different between the bile of cholesterol stones without biliary infection and

that of control group. Relati¥'e、decreasein cholic acid was observed in the bile with pigment

stones.

凪血－且血且

lO 20 Jllm且且

Z巴÷

Fig. 5 （；川ーliquidl、！日Jill日1けgramof a Test Mixture of Bile Aじid、

士一一寸一一一一~

一一一_j!_一一一一一一一ι-1!Lーι 一一一五車 革且；...

Fig. 6 C,1,-liquid ぐlirom九togramof Bile λcicトin HumanじけllbladderBile. C出 e1

Peptiに Ulcer

28 日本外科宝函 第3』巻 :xii －~； －

一一一一ーーーーーー『ーーーー－ ＿..ニニ丘三 一ーーーーーーーーーー←←一一一 『ー－

0

－ Fig. 7

~

・＿，，＿ー

z一一 ー一τAミニーニーミニ29_ - 2宣a忘却」 －

Gas-liquid Chromatogram of Bile Acids

in Human ( ;:illbladder Bile

Ca,;・ 5 : Cholesterol Stone

－‘

芳十十村山一；ー

ニii三

－：一三7弐之主Gas-liquid Chromatogram of Bile Acicb

in Human Gallbladder Bile

（．川、11: Pigment Stone

Table 8 G.1」liquidChnHll:ll＜収印phi・ of Human Bile λCl<b

Patient

Case 1

Case 2

Case 3 [

（、川、e4 !

一ι：＇＂＇ " Case 6

Ca寸 7

Case 8

仁川首 9

C."e IO

Diagnosis

Peptic Ulcer

//

//

，，

Mean

Ch11le、ter11lStone without Infection

//

//

//

//

必r

Me.in J

l'.1片 11j Pi I Inf札‘ti《》n

Ca同 12 I

に；N• 13 i

九七.111

，， //

1、ot・

v

(mg/ml J Dど‘’x、： t‘lwn《’《ie《』λy . Chol.

73.1 I 1.98 : 3.34

83.1 1 : 1.06 : 0.91

Ti目1 I I : 1.08 1.46

76.9 ・ I : 2.97 : 2.21

77.6 I 1 : 1.75 1.98

－一一一二一一一一no

49.9

30.8

36.7

51.5

5U

-11.7

56.9

I l.2

23.8

41.6

DISCUSSION

I

I

I

I

1.19

1.96

1.63

0.62

3.70

コ＿61

1.96

1.33

I I : 1.88

1 1.19

! I : I. I 0

0-96

2.03

1.58

1.00

~.79

3.71

'.2.01

1.59

1.62

1.26

1.12

入！thoughmany informations on the mechanism~

reported, the etiology of gallstone still remains obscure.

of氏allstone formation have been

EFFH :T OF EF A ON THE F< JRMATION OF ＜；λLLSTC>:'¥ES w

So far川じholesterolstones are concerned, it is reasonable to assume that cholesterol

solubility would be the most important in the formation of gallstones31l It has ・been

demonstrated by many investigators that bile acid and phospholipid (lecithin) which are

present in bile are important to keep cholesterol in solution12J21J22>2sJ3oJ Further, lsAKS-

SON23l ha日 demonstrated that a lecithin-bile salt complex plays the actual mechanism to

keep cholesterol in solution in human bile. This observation has been confirmec by the

solubility studies of JOHNSTON and NAKAYAMA.24> In our country, ;¥'lrYAKE34l ha-; also

shown that the ratio of lecithin to cholesterol should be more than 6.6 and at the same

time the ratio of B/C should be more than 8.0 to protect chole~terol precipitation in bile.

In general, bile acid and lecithin, which are important for holding cholesterol in solu-

tion, are decreased m礼rkedlyin the bile with gallstones. M1YAKE33l34J35> obsen-ecl marked

decrease in total bile acid, appearrance of free bile acid, relati¥・e incmζise in dihydroxy-

cholanic acid and decrease in lecithin and he emphasized that biliary infection was an

important factor for these changes. It has been reported by several im・estigators that selec-

tive change in permeability of gallbladder wall due to infection and inflammation lead to

the decrease of bile acid and lecithin and on the contrary, the increase of chole3terol in

bile42l51l. Thus it is generally accepted that the regional factors叩 chas infection, inflam-

mation, bile stasis and so on are responsible for cholesterol stone formation. On the other

hand，恥1ATSU032lemphasized the importance of systemic factors on the formation of gall-

stones. Further, ANDERSON1J has described that pure gallstone只 aredue to disturbョncesof

metabolism or of liver function rather than to an inflammatory reaction and that pure

gallstones may form the nucleus of the combined gallstoneべ. The results in the present

study show decrease in bile acid and lecithin, though biliary infection is not present. They

indicate that the composition of the bile with gallstonり when unassociated with biliary

infection is different from that associated with biliary infection. When accompanied with

biliary infection, decrease in bile acid and lecithin are more striking than when without

biliary infection and, in addition, qualitatiw change in bile acid and decrease in cholesterol

are observed. These facts indicate that the decrease in bile acid and lecithin in bile回 n

be occurred by some causes other than biliary infection. Therefore, it is nee引日n-to pay

attention to the importance of metabolic disturbances again.

It has already been demonstrated in our laboratory that cholesterol esterified with EFA

especially with tetraenoic acid is a precursor of adrenocortical hormones and that the ad-

renocortical capacitv of the individuals postulated in deficiencv or metabolic diぇturbanc回

in EFA are reduced15l. Al℃ording to these facts, HrKA.sλ13l has first surmised in 1960

that cholesterol esterified with tetraenoic acid should be the dirじじtprecursor of bile acid

and that deficiency or metabolic disturb礼ncesin EFA are responsible for gallstone formation.

It has heen report巳dby several investigators that highly unsaturated fatty acids from

vegetable oils promote cholesterol catabolism and bile acid excretion6l11l29l•1l44)_ Animal

experiment and clinical application in the present study show the same phenomenon also.

Bile acid in Ii＼＂げ ismarkedly reduced in the rat六 feela EF A -defi亡ientdiet, further, bile

acid and the ratio of B/C in hepatiじ bilein man are gradually incrt':tsl'cl lw the administr;i-

tion of both soya lecithin and pyricloxine. FuKUD.¥ 10l obsen-じda decrea町 inaclrenocortical

じapacityin patients with gallstones. HIRAN018l ohsen-ed that in the patient~ど bile with

30 日本外科宝函第34巻第I号

gallstones, fatty acid composition showed metabolic disturbances in EF A and that cholesterol esterified with tetraenoic acid was decreased. Further, YosHINAGA56> and HIRAN018> observed that decreased bile acid and lecithin in hepatic bile coincided with decreased cholesterol esterified with tetraenoic acid in liver were occurred in EFA-deficient rats. Therefore, cholesterol esterified with tetraenoic acid may be a direct precursor of bile acid. The biosynthesis of lecithin may be reduced in deficiency or metabolic dist町 bancesof EF A. As a matter of fact, lecithin is reduced in the patients’bile with gallstones. Recently, SmooA 45> succeeded in the production of cholesterol stones in hamsters, when they were put on an EF A-deficient diet for 2 to 3 months.

Cholesterol stone formation, however, is not due to the absolute deficiency in EFA, because the fatty acid composition of the bile with gallstones does not indicate EF A defi-ciency and the composition is not so different from that of control group. H1RAN018> also failed to observe EFA deficiency in the livピT of the patients with gallstones, he observed metabolic disturbances and disturbed utilization of EF A.

It has been reported by many investigators that pyridoxine is involved in the metabo-lism of fatty acids26> 21> 35＞叫. In accordance with these reports, bile acids were analysed with the liver slices of another group of rats which were fed a diet containing enough linoleic acid under deficiency in pyridoxine. Although any significant difference was ob-served between EF A diet and pyridoxine-def icient EF A diet groups in resting state, marked decrease in total bile acid in the latter was observed when the animals were administered ACTH-Z. Thus the composition of bile may be aggravated to pathological state by re司

peated exposure to stresses and finally, cholesterol precipitation may occur. It is now generally agreed that pyridoxine is involved in the metabolism of amino

acids and fatty acids. Pyridoxine deficiency syndrome can be observed when individuals are fed a diet high in protein＇則的 and saturated fat26>27>. This is particularly evident when the diet is high in methionine and cystine7> B> 43>. Production of pyridoxine by the intestinal flora is reduced with animal foods containing a large amount of protein and fatO叫. In general, animal fat and protein contain less pyridoxine than vegetables. Thus animal foods dieting are liable to pyridoxine deficiency. Actually, ScHROEDERrn reported that the American diet, containing a large amount of animal fat and hydrogenated vege-table fat and protein, might possibly be marginal with respect to this vitamin. As a matter of fact, gallstones in European and American compose chiefly of cholesterol. Recently the frequent occurrence of cholesterol stones has been observed in the city dwellers in our country who are apt to take a diet high in protein and animal fat with higher standard of life川. Moreover, it is interesting that they meet with stresses more frequently than countryman.

In accordance with these facts, it is evident that the metabolic disturbances in EF A due to pyridoxine deficiency play a very important role upon gallstone formation especially that of cholesterol stone. The total process of cholesterol stone formation in man is ii・lustrated in the schema described by HIKASA et al.11> in 1964. It should be emphasized that the initiating factor of cholesterol stone formation is attributed to the systemic one.

EFFECγ OF EFλ ti>: THE F(JJ＇入！.＼ Tit>>:OF ＜；礼LL討rtJ＞：！二円 31

V! SU!¥O!ARY主NDCONCLCSION

1) Bile acid and lecithin were decreased in the bile of patients with cholesterol

stones, though biliary inf町 tionwas not present.

2) The composition of the bile with cholesterol stones in the t、asesof without biliary

infection was different from that with biliary infection.

3) Therefore, changes in the bile with cholesterol stones may not be caused by m-

fection.

4) The resting levels of total bile acid in the rats' liver were clearly higher in EFA

diet and pyridoxine-deficient EFA diet groups than in EF入－deficientdiet group. After

the administration of ACTH Z, however, the levelメ oftotal bile acid reduced markedly

in pyridoxine-deficient EFA diet group.

5) The ratio of bile acid to cholesterol in human hepatic bile was elevated gradually

by the administration of EFA and pHidoxine.

6) Cholesterol esterified with EFλIll町’ bethe direct precursor of bile acid and

pyridoxine may be involved in the biosynthesis of bile acid from cholesterol.

7) The metabolic disturbances in EFえ dueto pyridoxine deficiency play a very

important role upon gallstone formation in man and stress affecい onthis process.

8) Bile acids from human bile were analysed by gas-liquid chromatography.

The author wi,hes to express hト叶日ceregratitude to .＼，，トLmtl'rけfeトト＇＞ rY ORP,ORI HI KASA for his gene1りLIS

guidance throughout the仁ourseof th卜 experiment,and to Dr. M1cH10 YosHI:-JAGA for his kind ccx1perat1リn.. '¥

part < ,f this ＂け rkwos reported at 63rd‘md 64th General Meeting of Japanese :-imgにal:-;, >ctel)

REFERENCES

1; .¥nder刈 >11, ¥¥" .• ¥. D. : Pathol＜収y,:-i1. Louis, 1957守 c.＼！.λJ,,,J" Co.

2) Bergstroem, :-i and ＼け1man .. ＼.：孔It・L 1 IJ, ,J ic product、ofcholesterol in bile and l町朴りfrat. Proc. ~凡 Exr》

Biol. Med., 83: 71. 1953.

3) Bloch. K., Bng. B.i¥. and Rittenberg, D. The biol＜収1c:ilconversion of cholesterol to cholic acid. J. Biol Chem., 149: 511, 1943.

,1) Blomぉtrand.R. and Ekdall, P. : F arty acid pattern <>f hum2111 bile under normal and pathological conditions.

Proc. ~当凡 ［ミ＇f〕 Biol. Med., 104 ： ~lJ5, 1960. 5 J Byers. S. () and Biggs, .¥!. ¥¥i. : Cholic acid and cholesterol. Studies concerning p＜同ibleintercon¥'er山川 λrch.

Hi・凡lw11LBiophys., 39: 301, 19云'.2.6) Bye＂＇・同 O and Friedman, H. : Bile acid metabolism, dietary fats, and plasma cholesterol le、el、Proc.S >c.

Exp. Biol. Med., 98 : 523, 1958 7) Cerecedo, L. R., Foy, J. R. and De Renzo. E. L、； Proteinintake and 、itaminB6 deficiency in the rat. The

effect of supplementing a le川一protein,pyridoxine-deficient diet with cystme or ¥¥'ith methionine. λrclL Bio-

chem., 17 : 397, 1948. 8) De bey, H. l，吋nell.E. E. and B:1umann, C.人： Studies<>I the interrelationship between methionine and

vitamin Br,. J. :¥utrition, 46 203, 1952. 9) Ellin, R. !.. i'Vlendeloff, .~. I. and Turner, D.λ ：Quantitative determination of ::. 7. 12-triket叫 h0Lin1cacid

in biological fluids by gas-liquid chromatography. 主11.il、ticalBiochem., 4: 198, 1962.

10) Fukuda, H.: personal communication. 11) Gordon, H., Lewis, B., Eales, L. and Broch, J. F .. IJwLcr、fatand cholesterol 1m・L1h<》！ism. Faecal ehm1即日

ti<>n of bile acid, and other lipids. Lancet, II : 1299, I日51.

12) Hammarsten, 0. : Zur Chemie der ( dle. Ergeb. Pin・叶什I,.4:1, 1905.

13) Hikas:c. ¥' et al.; Intravenous administration of a fat emulsion. (in Japanese) Ceka-shinrvo, 2 : 1650, 1960.

14) Hik＜ζ＂＇＂ Y et al.: :¥utritional efkct .,f Lit. (in Japanese) :-iogり l山 l札I.19 : 95, 1962. 15 i Hikasa, Y t'l al.：ドract1c.1lappli山 tionof gas-liquid chromatographic analy、トりlfatty acid,. (in Jar日ll<刊 l

32 日本外科宝函第34；会第 lE子

~.1i>hin-igaku, 18 : 921. 1963.

16) Hil、礼子、・1.Y et al. : ！~oles of EF A on adrenoc1汀ti印 lcapacity. (in .hp.111ee I Niho日イinshci,22: 142, 1964.

17) Hik'" 1. Y. et al. : Initiating factけ＂＇ of gallstones, especiallνcholesterol stones. Arch. Jap. Chir., 33 : 601,

196-1

18) Hirano, Y : personal communi印！1011.

19) Hir町 h,J田山1dλhrens,E. H. : The separation of complex lipid mixture lηthe u日 of、dicicacid chrリmatu-

graphy. J. Biol. Chem., 233 : 311, 1日S8.

20) Ikega山I,:¥. • （】山ーliquidchromatographic analv'i' of steroids and alkaloids. (in Japanese：ステロイドおよび

アルカロイドのガスクロマトグラフイー） K i芦北u-no－η6iki.15：」49.1961.

21J I、1ksson. B. : On the Ii p1d c1川、t1tuentsof normal bile .. .¥eta ~1c. Med. Ups1l., 56: 177, l日記

22) Isak弘明n,B. : On the lipid constituents of bile from human gallbladder containing cholesterol gall山川れ...¥eta

？ぢt凡 Med. Upsal., 59 : 2Ti, J 951

23) lsabson, B.: On the dissolving p1川町付tlecithin and bile salts for cholesterol in human bladder bile.弘・ta

ぬ1c.Med. t1p泊 I., 59: 296, 1954

24) Johnston, C. G. and l'¥akayama, F.：冷》l11bil1t、ofcholesterol and gall、tonれ inmetabolic material. . .¥. M. A

Arch. ~11r日， 75: 436. 1957.

251 K11収、l円。 G.R. and Schaffert, R.R.: Determination ol free and total cholesterol I"・ direct chlυn1!1》rmc~trac­

tion. J. Biol. Chem., 180 : 313, 1949.

26) Kotake，ミ： Triptophanmetabolism and experimental diabetes. (in Japanese：トリプトファン代謝と実験的

糖尿病lPr叫. J 6th General A出 embly]ap. Med. Congre田， 1: 299, 1963.

27) ｝い七1ke,Y. : Recent problems on the fat-nutrition. (in Japanese; I］訂1栄養における11立近の問題）片山.16th

General入、吋mblyJap. Med. Congre出， 1: 450, 1963.

28) Large, . .¥. M.: On the f，川n;i¥11111of日dlstones.吋ur伴、円。 54: 928. 1963.

29) Lewis, B. B. : Effect of certain dietan・ けils on bile acid s町 reti1111and世 rumch＜》bt℃ml.Lancet, I : J 090,

1958.

30) Lnng, J. H.川 dC~ephart, F .. On the behaYior of lecithin with bile salts and the occurr閉 じeof ll'l'ithin in

bile. ]. ..¥ m. Chem.冷凡， 30: 1312, 1908.

31) Lutton, R. G. and Lar耳e，主 M.. Gallstones：冷1lubil1t、、tudies.Surger¥・, 42 : 488. 1957.

32) Matsuo, I. : Galbtorws and diseases of bili川 tracts. (in Japanese：胆 （1と胆道の玖忠I19～17, Daigadり L、リ・

33) Miyake, H.: (di叫＜＞Ill.'di弓回目. !in Japanese) N1hけngeka-;-ensho, To川町 1957, Nankodo l、。34) Miyake, H. : On the fけrmation＜》igallstones. ( 111 Japanese ．胆fi生成機序に関して） Saishin-igaku, 14 : 3088,

1959.

35) Miy《;ike’H.:Ertl叩《＇Ile、l、andther;

λ、、t泊I】1bl、J之lf】. Med. C《》n耳ress’I: 733, 196 3.

36) Mosb山 h.E. H，κilin、h.H. J.. H"lpern. E. "nd Kendall, F. E. : Determination "f de＇山 vcholicand cholic acids in bile.λ札 11.Bi＜》chem.Biophys .. 51 : 402, 1954.

37) :'vluraoka, R. : Experimental study on the role of essential fatty acids and pyrirlリxineon adrenocortical function. λrch. Jap. Chir., 34 : 35, 1965.

38) ¥lueller, J. F. and L1co11<>. J. M.: Effect of desoxypyridoxine inducεcl vitamin BG deficiency口口 polyuns"turated

fatt、acidmetabolism in human beings.主m.J. Clin. :¥utr .. 12: 358. 1963.

39） 川、himura,M. : Ueber die Chemische Zu泊 mmensetzungder Gallensteine. ]. Bi叫 hem.,28: :!65, 1938.

40) :--iiトhimura.M. : Gas-liquid chrom 1tographic anal、sisof fatt)' acids in human bile. (in J,1p.1nt・st・ ：ガ 7..7ロ7

｝グラフイーによる人胆作中脂肪酸の分析）吋ιi山 m11 I Oji’K and 九1ι1t 、 11'.1~ 片： Essenti凶l Litt¥・ 《aci《band cholesterol 口1eta lism. I in Japa11円．：！ι、須脂肪駿とコレ

ステロール代謝）Sog6-rinsh6, 7 : 2296, 1958.

12) Iミiegel,C., Ravdin, IS and Johnston, C : Studies of gallbladder functiり日 VI. ’！＇he absorption of bile s;dts

and cholesterol from the bile fr什 gallbladder.Am. J. l'll¥siol., 99・656,1932.

43) Sakurai, Y ;111d Hdra, H.: Effeじtof sulfur-amino acids けn vitamin 86・（ inJapane問： 136作用に；＆11 す；＇~Mcアミノ般の影特j Vitamin, 5 : 415, J 952.

-1・1 J Sat<>. J.: Stu凶作 onthe el i町ト口nun川 turated fatty acid administration upon biliary components. (in Japanc、町）J. Kurnme Med. Assoc .. 24: 1800, 1961.

」五） ＆・h口町ler,・H.λ : Is .ithen1sderosis a cりnditionedpyndoxal def1cimn・? J. Chron. !Ji,., 2; 28, 1955.

16 I Schweigert, B. S., S出 1berlich,H. I、..Elvehjem, C. A. and Baumann, C.九： Ilie七1r、proteinand the v1tan11n

Bりじりntent"I mou田 ti出ue.J. Biol. Chern., 165 : 187, 1964.

EFFECl‘ （）ト EFλ < J~ THE FOR:¥1λTl< J:¥ （）ド <,.¥LLST<J:¥I二円 33

171 Sheppard, E.仁川idMc Henry, E. ¥¥' : The p、ride山 inecontent "f ti同 LI（ヘりfrats fed various dieb. ] . !),, ,J

Chem., 165 : 649, 1946.

48) 吋l11≪L1. l~. ： pc円汁1,ilcommu111山 tion.

lり） Si per吋tein,M. D. and Chaikoff, l. L.: c11 chol凹 tern!.Ul. Ex仁 川lりnof carbつns4 an仁126 in l印刷 Urme

and bile. J. Biol. Chem .. 198: 93, 1952.

SO I 吋J＂円 .ill.J.: qu;dit川1、ぞ礼mlysisof bile ;;cicb. by gas chromat<>grιq>ll¥ Bile acJCト andsteroids 1 2 l. 主山iChem.

Sc・;ind .. 16 : 1761, 1962.

51) Uraki, ] . : Bile and choleメter口I.(in ] a pan出 e：胆汁とコレステロール）初日（）－］＇］］ト lio.7 : 2255, 1958.

521 ¥V；ι1tanabe, N.: F.1tt1 acids in bile. (in ]a panesぞ： 1J11?Iの！日ilj俄＇ Nihon fllbhO, 22 : 541, 196,l.

531 Witten, P. ¥¥' and Holman. R. T.: Pulyethenoid fatt1 acid rn山 1boli叶n.¥'!. El iιlりlpyridoxine on o町 nt1al

fatty acid rnm・ersiけ日． :'ucli. B1< 、hem.. 41 : 266, 1 ~Ei2

5.1 I Yano, M : Elkeト ofc・cllulc町 uponthe srnthesis of vitamin ll,, by intestiml flora. I in Japa即日：人体iU,Jil~

細菌によるBsの合成とセルロースの影特 1Vitamin, 10 : 166. 1956.

詠） Yoshida, l¥l. .ぐlit11仁社IStud日、 ongallstone disease. (in J日 l川町：胆百症の叫ん•qミ的研究 l ]. Jap. Smg. Soc.,

60: 599. 1959.

56) Y"shimg;1, M.: Experimental studies '"' the initiating factor rd ιhc,Jり lLTCJi gall山川氏、， especiallyon the in-

fluence of essential fott1 ιi仁川、 andpyridoxine on the bile constituεnts. 入＇＂I' Jap. Chir., 34 : 1 .. 1965.

34 日本外科宝函第34巻第1号

和文抄録

不可欠脂酸及びビリドキシンの胆石，殊にコレステロール系

結石の形成に対する意義

京都大予医学店外科学教室第2講座（指導：木村忠司教授）

丸

胆石症の成因に関して： t, :¥aun¥'n及び A"'-・ho ft以

来，内外にlj'c:て幾多の研究業績が発表さ1.tて来たが，

現庄尚不明の！.＇！， ;j；多いー従来，一般に小ld感.；~，胆汁

鰐滞等の局所性因子戸重視されて来たがp 胆道感染の

認められない純コレステロール結石や，無 （j胆嚢炎等

コ1日常の臨床に於てEID;々 みられる点からすると p 胆石

症の成因を局所性因子のみで説明する事は出ういない．

従って，新陳代謝の ＇ii;：：；；；，内分泌機能の＇ J;:,・r;,J，肝機能

の不全等の全身性要因を重視する人もある．我々の教

室ではp 日笠等が夙に干三可 t、lj[了，，建

に；，＇，日し P そのif伏Lこつとめて来たがp 最近に至り p

副腎皮質ホルモハけに Gluc叫 orticoidの前駆物質はp

不可欠II旨酸， 殊に Tetraenoicacidと＿［＿ ステル結合し

たコレステロールであり， 不ロI欠JJ旨酸欠乏時にはp

副腎皮質ホルモンの産主能が低下する事を知り得るに

至った． 4 方，コレステロールの終；lミ代謝必物ii胆汁

酸てある点i二郎Lみ，胆汁酸の生合成に阪しても，亦コ

レステロールは不可欠！日限とエステル結合していなけ

ればならないということが充分推測され仰る．このよ

うな汚え方からすjtli，不可欠I］汁燃の欠乏P 乃至はそ

の代謝障害の{j.在する場合にはp 当然胆汁酸の産生は

障害され，胆汁中への胆i'I酸分泌能も低下し， これが

胆汁の不安定化を招き，結石1ド1A：：重大な配管を及If

すものと考えられる．そこで不可欠脂酸史的その代

謝に重要な役割号果すP ピリドキシンの胆r！車ll口！＜，に及

ぼす影響をみるためにp 以下のような臨床的及びy 基

liJ}!的実験壱施行した．即ちP 手術時無菌的に採取し

た，！但 （j症患、者交ぴ，対照群としてF 消化性潰疹患者

の胆嚢胆汁を夫々分析す石と同時にp 更に総胆管ドレ

ナージを施し九皆、者にp 不可欠脂酸及びピリドキシン

を投与してP 肝JJ旦汁組成の変動を紙日的に分析，検討

した． 更にy ウ fスター系雄性ラットを不可欠II旨酸

’hu円’tr

泉

食，不可・.. :im般に之ft，ピリドキシン欠乏・不可欠脂

酸食，にて夫町 2～3ヵ月に互り飼育し，その肝臓

に含有される，阻戸｜酸をも分析し次のよ弓な結果を得

fご．

ill コレステ U-'c系私！？石患者の胆嚢胆汁ーではp 肝

機能障害や胆道感染が認められなくても，総胆汁酸量

及びレチティン量の減少が認められる．

(2) 胆道感染の認められないp コレステロール系結

石患者の胆汁組成は，胆道感染の認められるコレステ

ロ＿，，系結L1患者のそれと，明らかに異っている．

13) 従ってP コしステロール系紡（Iにみられる胆F卜

組成の変化は，九銭＇｛＇ Iこ起｜人｜するものとは考え難い一

i・ll ラ、ット肝臓中に含有される ljll1f限量は，安静時

に於て，不＂J• .. :Iii:!再建欠こ fr/ffでは皆I＇•＇~している．一方P

1トしスを加えると p 不可欠IJ旨酸食群ではその肝臓合

有胆汁酸量に皆変ボ認められないのに対し，ピリドキ

シン欠乏・不可＇ rll'1l限ttmでは；七ihiQ;j＞認められた．

(5) 総胆管ドレナージを砲した患者の肝胆汁では，

不可欠脂酸見ひピリドキシンの投与により，胆汁酸量

点噌加しp 胆f卜酸対コレステロールの比IBl（、比）の上

昇が認められた．

161 共同研究ん ，＇｛永p 平野等の実験成績を併せ考

えると＇ ij旦汁酸の前駆物l'tlt，不可欠脂酸，殊にTet-

raenoic acidとエステル結合したp コレステロ＿，，であ

ると廷えらjLる．

(7) ピリドキンン欠乏に起因する不可欠町般の代謝

障害はp 胆 Li，殊にコレステロール系結行の形成に重

要な役割を果しP 1トレスがこれを助長するものと考

えられる．

18) 人胆汁中に ：~ イJされる胆汁酸をガスクロマトゲ

ラフィーでうJtlrする：・J；にも成功した．