Effects of Lycii Fructus on the Ovariectomized Osteoporosis of Rats

서 론

(bone matrix) , (bone mass)

1)

. ,

,

.

枸杞子 卵巢摘出 骨組織代謝 影響

구용모

, 김성훈

, 김은영

, 이현삼

, 최현

, 손영주

, 정혁상

, 손낙원

Original Article

Effects of Lycii Fructus on the Ovariectomized Osteoporosis of Rats

Yong-Mo Koo ¹ , Sung-Hoon Kim ² , Eun-Young Kim ¹ , Hyun-Sam Lee ¹ , Hyeon Choi ⁴ , Young-Joo Sohn ³ , Hyuk-Sang Jung ¹ , Nak-Won Sohn ²

1

Department of Anatomy & Institute of Oriental Medicine, College of Oriental Medicine, Kyung Hee University,

2

Department of Neuroscience and Brain Disease, Graduate School of East-West Medicine, Kyung Hee University,

3

Department of Gynecology, College of Oriental Medicine, Sangji University,

4

Hamsoa Pharmaceutical Co., Seoul

Objectives: Osteoporosis is one of the most common metabolic bone disorders in the menopausal stage, a disease signified by decrease of bone mass. The purpose of this study was to learn how Lycii Fructus influences SD rats with osteoporosis caused by the removal of the ovaries.

Methods: After the removal of the ovaries from 24 female Sprague-Dawley rats, they were divided into a normal group, a control group, and a Lycii Fructus (25mg/100g) administration group. After 8 weeks, the rats were sacrificed and their weights, albumin, AST, ALT, ALP, total-cholesterol, triglyceride, P, Ca, T4, E2, weight of a femur, ash content of the tibia, and area and thickness of trabecular bone were measured.

Results: Regarding the density of ALT, ALP, and estradiol from serum, the Lycii Fructus group was significantly decreased compared to the control group, and the density of phosphorous and calcium also decreased slightly.

Moreover, albumin, triglyceride, and T4 showed an increasing propensity, but AST and total-cholesterol showed a decreasing propensity, but not significant.

Regarding the variation of bone, femur weight and ash content of tibia, the Lycii Fructus group was increased compared to the control group but significance wasn’t shown. The Lycii Fructus group increased significantly over the control group in area and thickness of trabecular bone and osteoclast number.

Conclusions: From the results of the above study, Lycii Fructus should be effective for osteoporosis treatment and prevention.

Key Words : osteoporosis, ovariectomized rat, Lycii Fructus , trabecular, osteoclast

2008 6 2 2008 7 2

: , 1

(Tel : +82-2-961-9449, Fax : +82-2-961-0327, E-mail : [email protected])

( ) R&D BK21

.

This research was supported by Hamsoa pharmaceutical

Co. R&D Center and BK21 Project, Republic of Korea.

.

, ,

, ,

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.

2)

. (Solnaceae)

Lycii Fructus

3)

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4-5)

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,

. Saville

,

6)

,

7)

.

,

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, osteoclast ,

calcium (Ca), phosphorus (P), tetraiodothyronine (T4), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), albumin, totalcholesterol, triglyceride, estradiol

재료 및 방법

1.

동물

Sprague-Dawley(SD)

. 170g 21-

23 , 40-60%, 12

,

1 200g

.

2.

검액의 제조

.

300g 3,000ml

2 .

, vacuum rotary evaporator 26.1%

.

3.

실험군의 분류

SD-rat

. ,

,

,

8 .

4.

골다공증의 수술 및 검액 투여

350mg/kg chloral hydrate

. ,

. 2 8

1 100g 25mg/ml

,

.

5.

체중 측정

2 8

.

6.

채혈 및 혈액분석

8 SD-rat

2,000rpm, 4

10 . albumin,

AST, ALT, ALP, total cholesterol, triglyceride, T4, Ca, phosphorus, estradiol

.

7.

대퇴골 무게와 경골의 회분 함량 측정

8

,

900 24

ash .

8.

조직 화학적 염색

neutral buffered formalin (NBF)

1 , , 2

formic acid ,

. 4 um

hematoxylin and eosin (H&E)

40 , 100 .

9.

대퇴골 조직의 관찰

H&E 40 capture

2/3

0.4126702×107

NIH image

1.62 ,

. Osteoblast 100

capture 0.1511111×107

NIH image 1.62 .

Osteoclast

2/3 200

.

10.

통계처리

SPSS® for windows (version 10.0, SPSS, Inc., Chicago, U.S.A.)

oneway ANOVA ,

P value<0.05

post hoc test Scheffe's test .

실험성적

1.

체중 변화 8

(Normal) 214.50 ± 5.95g 282.75 ± 9.71g (OVX) 205.29 ± 1.43g 335.86

± 7.73g (GGJ) 206.63 ± 1.40g

346.38 ± 4.86g .

, (Fig. 1).

2.

혈액성분의 변화

1) Albumin

albumin 4.10 ± 0.12(g/dL), 3.66 ± 0.07(g/dL)

(P<0.05), 3.69 ± 0.07(g/

dL) (Table 1).

2) AST

AST 115.25 ± 16.98(IU/L), 121.71 ± 14.89(IU/L), 100.13 ± 4.11(IU/L)

(Table 1).

3) ALT

ALT 58.00 ± 13.06(IU/L), 52.57 ± 5.62(IU/L), 41.25 ± 2.36

(IU/L) (P<0.05)

(Table 1).

Normal OVX GGJ

Albumin (g/dL) 4.10 ± 0.12 3.66 ± 0.07# 3.69 ± 0.07

AST (IU/L) 115.25 ± 16.98 121.71 ± 14.89 100.13 ± 4.11

ALT(IU/L) 58.00 ± 13.06 52.57 ± 5.62 41.25 ± 2.36

ALP (U/L) 184.25 ± 32.07 179.71 ± 16.87 137.75 ± 10.19

Total-cholesterol (mg/dL) 80.50 ± 3.48 108.29 ± 7.03

97.25 ± 3.71 Triglyceride (mg/dL) 114.50 ± 14.54 39.57 ± 3.25

50.38 ± 5.78

P(mg/dL) 6.23 ± 0.15 5.26 ± 0.35

7.14 ± 0.16

Ca(mg/dL) 10.75 ± 0.12 10.06 ± 0.16

10.58 ± 0.12

T4 ( ㎍/dL) 1.35 ± 0.06 1.83 ± 0.14

1.84 ± 0.10

Estradiol (pg/dL) 14.73 ± 0.30 9.30 ± 0.23

8.39 ± 0.36

Normal; non-treated group.

OVX; administrated with D.W (1ml) daily by oral gavage for 8 weeks in ovariectomized rats.

GGJ; administrated with Lycii Fructus (25mg/ml) daily by oral gavage in ovariectomized rats.

Data are expressed as mean value ± S.E

# P<0.05, ## P<0.01 , ### P<0.001 : significantly different from normal group.

* P<0.05, *** P<0.001 : significantly different from OVX group.

Table 1.

B od y w eig ht (g )

360 340 320 300 280 260 240 220 200 180

1 2 3 7 10 14 17 21 24 28 31 35 38 42 45 49 53 56

daysafter surgery

Normal OVX GGJ

Fig. 1.

4) ALP

ALP 184.25 ± 32.07(U/L), 179.71 ± 16.87(U/L), 137.75 ±

10.19(U/L) (P<0.05)

(Table 1).

5) Total-cholesterol

Total-cholesterol 80.50 ± 3.48(mg/

dL), 108.29 ± 7.03(mg/dL)

(P<0.01) ,

97.25 ± 3.71(mg/dL) (Table 1).

6) Triglycerice

Triglycerice 114.50 ± 14.54(mg/dL), 39.57 ± 3.25(mg/dL)

(P<0.001) , 50.38

± 5.78(mg/dL) (Table 1).

7) Phosphorous

Phosphorous 6.23 ± 0.15(mg/dL), 5.26 ± 0.35(mg/dL)

(P<0.05) , 7.13

± 0.16(mg/dL) (P<0.001)

(Table 1).

8) Ca

Ca 10.75 ± 0.12(mg/dL),

10.06 ± 0.16(mg/dL) (P<

0.01) 10.58 ± 0.12(mg/

dL) (P<0.05)

(Table 1).

9) T4

T4 1.35 ± 0.06( /dL),

1.83 ± 0.14( /dL)

(P<0.01) 1.84 ± 0.10

( /dL) (Table 1).

10) Estradiol

Estradiol 14.73 ± 0.30(pg/dL), 9.30 ± 0.23(pg/dL)

(P<0.001) , 8.39 ±

0.36(pg/dL) (Table 1).

3.

대퇴골의 무게

1.572 ± 0.030g, 1.674 ± 0.049g, 1.737 ± 0.037g

(Table 2).

4.

경골의 회분함량

0.2524 ± 0.0048g, 0.2647 ± 0.0087g, 0.2773 ±

Left femur(g) Right femur (g) Entire femur (g) Tibia ash weight(g)

Normal 0.786 ± 0.009 0.786 ± 0.021 1.572 ± 0.030 0.2524 ± 0.0048

OVX 0.842 ± 0.026 0.832 ± 0.024 1.674 ± 0.049 0.2647 ± 0.0087

GGJ 0.875 ± 0.017 0.862 ± 0.021 1.737 ± 0.037 0.2773 ± 0.0046

Normal; non-treated group.

OVX; administrated with D.W (1ml) daily by oral gavage for 8 weeks in ovariectomized rats.

GGJ; administrated with Lycii Fructus (25mg/ml) daily by oral gavage for 8 weeks in ovariectomized rats.

Data are expressed as mean value ± S.E

Table 2.

0.0046g

(Table 2).

5.

골소주의 두께

118.61 ± 5.31 , 96.23 ± 2.32

(P<0.01) 114.04 ± 2.89

(P<0.001)

(Fig. 2).

6.

골소주의 면적

71.14 ± 3.35%, 56.02 ± 0.47%

(P<0.001) 63.90 ± 1.06%

(P<0.001) (Fig. 3, 4).

##

150

Tra be cu la r t hic kn es s (u m )

Normal 0

120 ***

90

60

30

OVX GGJ

Fig. 2.

###

80

Tra be cu la r a re a (u m )

Normal 0

***

60

40

20

OVX GGJ

Fig. 3.

7. Osteoclast

의 개수

Osteoclast 0.37 ± 0.07 (No

/mm), 0.56 ± 0.07 (No/mm) , 0.24 ± 0.03(No/mm)

(P<0.001) (Fig. 5).

8. Osteoblast

의 면적

Osteoblast 2.06 ± 0.18%,

5.25 ± 0.81%

(p<0.01) , 2.60 ±

0.09% (P<0.01)

(Fig. 6).

1 2 3 4

5 6 7 8

Fig. 4.

0.7

O ste oc la st (N o/ m m )

Normal 0

***

0.6

0.4

0.2

OVX GGJ

0.5

0.3

0.1

Fig. 5.

고 찰

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9-10)

.

, , ,

estrogen , Vit.D , calcitonin

11-13)

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14-15)

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estrogen, calcitonin , osteoclast

bisponate

, fluoride

. estrogen

, , ,

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18-19)

. Saville

,

6)

.

estrogen

, 8

albumin, AST, ALT, ALP, total-cholesterol, triglyceride, estradiol, phosphrous, calcium, T4

, , osteoclast

. Albumin

60% . Albumin

Ca, P, S

7

O ste ob la st (% )

Normal 0

**

6

4

2

OVX GGJ

5

3

1

##

Fig. 6.

, albumin

20)

. albumin

, . AST, ALT

, , ,

. AST

, ALT

AST

20)

.

AST, ALT ,

AST , ALT

.

. ALP

, . ALP

,

22)

. ALP

.

Estrogen estradiol, estron, estriol 3 , estradiol

estrogen ,

estrogen

23)

, estrogen

,

. cholesterol

triglyceride cholesterol

LDL-cholesterol cholesterol

16)

.

. total-cholesterol

, triglyceride estradiol

. total-cholesterol

estradiol ,

triglyceride .

Calcium 99.9% 0.1%

.

calcium calcium

,

calcium

24)

. Phosphorous 85%

. phosphorous , ,

20)

,

calcium calcium

phosphorous phosphorous

calcium

.

Phosphorous calcium

, Phosphorous calcium

.

90% thyroxine, tetraiodoth- yronine(T4) 10% triaiodothyronine

,

.

6,26)

calcium

osteoclast

27-28)

.

, T4

,

.

estrogen calcium phosphorus

29)

. ash weight

30)

,

,

.

tibia ash

,

. osteoclast

, .

, ALT

, phosphorous, calcium, , osteoclast

.

.

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