Study of Effects and Optimum Concentrations of Medical Herb Extracts on Blood Glucose and Serum Lipid in Streptozotocin-Induced Diabetic Rats

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서 론

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2008 1 25

2008 3 14

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3 (Tel:+82-63-850-6836, Fax:+82-63-852-5594, E-mail:[email protected])

정명수

¹

, 김혜자

¹

, 조화은

²

, 최윤희

²

, 이기남

^2,3

Original Article

Study of Effects and Optimum Concentrations of Medical Herb Extracts on Blood Glucose and Serum Lipid in Streptozotocin-Induced Diabetic Rats

Myong-Soo Chong

¹

, Hae-Ja Kim

¹

, Hwa-Eun Cho

²

, Yun-Hee Choi

²

, Kin-Am Lee

^2,3

1

College of Oriental Medicine,

²

Professional Graduate School of Oriental Medicine,

3

Research Center of Traditional Korean Medicine, Wonkwang University

Objectives : The purpose of this study was to investigate effects of extract from mixed culture with Tricholoma matsutake mycelium and Cordyceps militaris mycelium in oriental medicine and cereal midium (OCM) considered effective for the treatment of diabetes.

Methods : We examined antioxidant activity and α-glucosidase inhibit activity of OCM and orally administered OCM extracts to Streptozotocin-induced diabetic ratsat the dose of 100mg/kg, 200mg/kg and 300mg/kg body weight.

Results : Total polyphenol content of extracts of OCM was 26.41 mg/g, DPPH radical scavenging activity was 32.24% and SOD-like activity was 31.43%. α-glucosidase inhibit activity showed 85.3% of acarbose. Blood glucose levels of the OCM extracts-treated groups were lower than those of the diabetic control group (DC), total cholesterol and triglyceride contents were similar to the acarbose-treated group (PC) and ALT, AST were lower than the DC and PC groups. The 300 mg/kg body weight administered group (UM-300) were superior to other OCM-treated groups (UM-100, UM-200).

Conclusions : These results showed extract from mixed culture with Tricholoma matsutake mycelium and Cordyceps militaris mycelium in oriental medicine and cereal midium has potentiality to beused in pharmaceutical products and medicinal food for preventive and treatment diabetes mellitus.

Key Words : Orient medicine medium, diabetes mellitus, lipid profile, mushroom.

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. 20~

70%

⁶⁾

,

, ,

7-9)

.

, ,

. (Tricholoma matsutake)

β-glucan

10-13)

. (Cordyceps militaris)

,

¹⁴⁾

. , ,

, ,

15-19)

.

β-glucan cordycepine

.

10 ,

(Oriental medicine and Cereal Medium;

OCM)

STZ(Streptozotosin)

.

재료 및 방법

1.

실험재료

2007

Table 1 . (Trichloloma

matsutake mycelium) , (Cordyceps militaris mycelium)

. (Lactobacillus plantarum 3108) (Saccharomyces cerevisiae 7268)

.

2.

균사체 배양

,

발효 및 시료의 추출

(3)

1.2 121

15 ,

5% 2

4 7 .

95 30

0.5% 24 36~48

(pH 4.0) 15 ,

,

(UMPM, MEDIPS, umex, Korea)

. ,

.

3.

항산화활성 측정

: Folin-Denis

²⁰⁾

1 mg/mL 2N Folin 200 μL

3 . 10 %

Na2CO3 2 mL 1

ELISA reader(Bio-Tek Instruments,

INC) 725nm

. tannic acid

. SOD : SOD Assay kit-WST(Dojindo Molecular Technologies, Inc, Japan)

20 μL WST-1 200 μL, Enzyme 20 μL , blank

37 20 450 nm

.

: Blois

²¹⁾

100 μL 4.0×10-4M DPPH (99% Ethanol)

150 uL , 30

525nm .

4.

α－

glucosidase

저해활성 측정

α-glucosidase 10 μg/mL

0.1 unit/mL α-glucosidase(Sigma, Co, U.S.A) 50 μL, 200mM PBS buffer(pH 7.0)

50 μL 37 15

3mM pNPG(Sigma, Co, U.S.A) 37

10 . 1M glycine-NaOH

405 α-glucosidase acarbose .

5.

실험동물 및 당뇨유발

6 Sprague Dawley ( )

Material Composition(%)

Hordeum vulgare var. hexastichon 15

Fagopyrum esculentum 15

Morus alba 14.5

Eucoimmia ulmoides 13.3

Glycyrrhiza urlensis 7.5

Liriope platyphylla 8.8

Dioscorea batatas 8.8

Panax ginseng 6.3

Trichosanthes kirilowii 5.7

Beauveria bassiana 5.1

Table 1.

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( ) (

22±2 , 50±5%, 12 )

stainless cage

. 16

β-

streptozotocin(STZ, Sigma. Co, U.S.A) 0.01M citrate buffer(pH 4.5) 50 mg/kg/body weight

. citrate

buffer . 24

300 mg/dL

. (NC )

(DC ) (PC ),

100 mg/kg (UM- 100

), 200 mg/kg (UM-200 ), 300

mg/kg (UM-300 ) 6

. NC DC 0.9% ,

PC acarbose 50 mg/

kg , UM-100, UM-200, UM-300

(0.9%, saline)

1 1 .

2 (AIN-93 diet)

.

6.

혈당측정 및 혈액

,

조직의 채취

3 (Accuchek

Mixed of OCM

Added with 1.2vol. D.W. (121 ℃. 15 min)

Inoculared with Tricholoma matsutake & Cordyceps militaris (5%) 24 ℃. 7days

｜

OCM culture broth

Autoclaved (95 ℃. 30min)

｜ Inoculated with

Lactic acid bacteria & Yeast Fermented (24 ℃, 36～46hr)

｜

Added with 15vol. D.W. (65 ℃, 4hr) 1)Micro wave (2450MHz) 30min 2) U.Sonic (26KHz) 30min 3) U.Sonic (65KHz) 30min 4) U.Sonic (128KHz) 30min 5) With micro bubble and vacuum

｜

UMPM fermented extract

｜

Centrifuged

｜

Evaporation

｜

Freeze dryin

Fig. 1.

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Active. Roche Diagnostics Cmb, Germany)

. 12

ether

3 3000rpm 20

. ALP AST, ALP,

, , creatinine, BUN, uric acid kit(Bayer, USA) (Advia 1650, Tokyo, Japan) .

4% NBF buffer(pH 7.2~7.4)

4 μm H-E(Hematoxylin

& Eosin) staining , .

7.

통계처리

SPSS 12.0 program

one-way ANOVA ±

, Duncan’s

multiple range test p<0.05 .

결과 및 고찰 1.

항산화활성

free radical

22,23)

.

SOD

Table

2 . 2

,

26.41 mg/g microwave

9.20 mg/g Lee

²⁴⁾

Lee

²⁵⁾

5.80 mg/g, 2.10 mg/g, 2.00 mg/g

. 1 mg/mL

32.24%, SOD 31.43%

.

2.

α－

glucosidase

저해활성

α-glucosidase

. acarbose,

voglibose α-glucosidase

Total polyphenols (mg/g) DPPH activity (%) SOD-like activity (%)

Antioxidant activity 26.14 ± 1.09 32.24 ± 0.90 31.43 ± 7.08

All values are mean±S.D.

Table 2.

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.

α-glucosidase

acarbose , (Fig. 2).

10 ug/mL acarbose 77.28%, 60.97%

acarbose 78.89%

.

acarbose 83.5% 52.1%

Shin

²⁶⁾

.

3.

혈당에 미치는 영향

Fig. 3 .

NC 107.20 mg/dL , STZ

Alpha-glucosidase inhibitory activity (%)

100

80

60

40

20

0

Acarbose UM

Fig. 2. α

Blood glucose (mg/dL)

700

600

500

400

300

200

100

0

0 1 4 7 10 13 15

(day)

NC DC PC UM-100 UM-200 UM-300

Fig. 3.

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, DC 513.75 mg/dL, PC 498.37 mg/dL, UM-100 513.71 mg/dL, UM-200

498.71 mg/dL, UM-300 496.71 mg/dL

. 2 acarbose

PC

, DC

537.37 mg/dL

UM-100 487.71 mg/dL, UM- 200 523.28 mg/dL, UM-300 469.00 mg/dL DC

UM-300 .

4.

혈청지질에 미치는 영향

2-5

²⁷⁾

70-80%

²⁸⁾

.

Table 3 .

29,30)

.

UM-100 77.0 mg/dL ,UM-200 83.71 mg/dL, UM-300 81.43 mg/dL

acarbose PC 74.00 mg/dL DC 104.20

mg/dL .

UM-200 UM-100, UM-300

UM-200

.

UM-300 96.71 mg/dL .

UM-300

25.9%, 36.0%

.

Group Total cholesterol Triglyceride HDL-cholesterol LDL-cholesterol AI NC 64.60 ± 6.54

^a

77.60 ± 21.25 21.80 ± 2.38

^a

27.28 ± 5.29 1.96 ± 0.13

^c

DC 104.20 ± 16.88

^c

151.20 ± 34.21 39.60 ± 5.27

^c

34.36 ± 7.64 1.62 ± 0.22

^ab

PC 74.00 ± 13.70

^ab

103.13 ± 74.86 29.38 ± 7.94

^ab

24.00 ± 10.23 1.57 ± 0.23

^ab

UM100 77.00 ± 14.68

^ab

108.86 ± 66.41 31.57 ± 7.82

^bc

23.65 ± 9.80 1.47 ± 0.22

^ab

UM200 83.71 ± 18.14

^b

125.57 ± 64.33 35.00 ± 8.71

^bc

23.60 ± 6.86 1.40 ± 0.13

^a

UM300 81.43 ± 14.55

^ab

96.71 ± 52.16 30.29 ± 5.58

^b

31.80 ± 9.70 1.69 ± 0.25

^b

NC: normal control group, DC: diabetes control group, PC: positive control group(treated by acabose 50 mg/kg),

UM100: UM extract(100 mg/kg), UM200: UM extract(200 mg/kg), UM300: UM extract(300 mg/kg)

All values are mean±S.D. Alphabet: Different superscripts in the table indicate significant difference by Duncan's multiple range test.

Table 3.

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5.

혈청 중

metabolic variables

의 변화 Table 4

. AST NC 176.00

U/L DC 440.20 U/L

PC 399.88 U/L UM-300

241.29 U/L, UM-200 247.00 U/L, UM-100

265.00 U/L PC . ALT

NC 37.40U/L

DC 171.20 U/L

PC 140.75 U/L

UM-300 79.57 U/L, UM-200 84.29 U/L, UM-100 96.00 U/L

. AST ALT

.

AST ALT

31)

.

AST, ALT DC

, PC

300 mg/mL UM-

300 . Creatinine DC

0.74 mg/dL ,

. BUN NC 20.40 mg/dL, DC 67.24 mg/dL, PC 36.10 mg/dL, UM-300 35.52 mg/dL, UM-200 34.94 mg/dL, UM-100 53.60

mg/dL , Uric acid NC 2.42

mg/dL, DC 2.76 mg/dL, PC 2.08 mg/dL, UM- 300 3.48 mg/dL, UM-200 2.68 mg/dL, UM-

100 1.74 mg/dL .

결 론

α-glucosidase

STZ(Streptozotosin)

.

26.41%, 32.24%,

SOD 31.43% α-glu-

Group AST

(U/L)

ALT (U/L)

Creatinine (mg/dL)

BUN (mg/dL)

Uric acid (mg/dL) NC 176.00 ± 34.05

^a

37.40 ± 9.91

^a

0.64 ± 0.05

^a

20.40 ± 2.30

^a

2.42±0.41

^ab

DC 440.20 ± 261.90

^c

171.20 ± 76.17

^c

0.74 ± 0.05

^b

67.24 ± 15.23

^d

2.76±0.43

^b

PC 399.88 ± 196.84

^bc

140.75 ±112.45

^bc

0.66 ± 0.07

^a

36.10 ± 9.29

^b

2.08±0.47

^ab

UM100 265.00 ± 85.19

^ab

96.00 ± 50.72

^abc

0.62 ± 0.07

^a

53.60 ± 9.56

^c

1.74±0.28

^a

UM200 247.00 ± 57.57

^ab

84.29 ± 12.17

^ab

0.60 ± 0.05

^a

35.52 ± 5.71

^b

2.68±0.59

^b

UM300 241.29 ± 64.67

^ab

79.57 ± 27.75

^ab

0.58 ± 0.06

^a

34.94 ± 5.82

^b

3.48±0.94

^c

NC: normal control group, DC: diabetes control group, PC: positive control group(treated by acabose 50mg/kg),

UM100: UM extract(100mg/kg), UM200: UM extract(200mg/kg), UM300: UM extract(300mg/kg)

All values are mean±S.D. Alphabet: Different superscripts in the table indicate significant difference by Duncan's multiple range test.

Table 4.

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cosidase acarbose 85.3%

. DC

,

acrabose PC

,

. ALT, AST DC

acarbose .

UM-300 ,

UM-100, UM-200 300

mg/kg AI

Uric acid

.

, .

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