ANH HUONG CUA POLYSACCHARIDE CHIET TU QUA THE VA SINH KHOI SOI NAM LINH CHI (GANODERMA LUCIDUM) NU(DI TRONG LEN TRONG LUONG VA THOI GIAN SONG THEM CUA CHUOT NHAT TRANG SWISS MANG U B A N G SARCOMA 180

N g u y e n Thi Quy, H o a n g Thi My N h u n g , D i n h H o n g D u y e n Khoa Sink hoc, TrUdng BHKHTN, BHQGHN

MODAU

Ung thU la mot can benh hiem ngheo vdi ty le t\i vong vao loai cao nhat tren the gidi. Ty le ngUdi mac benh hang nam ciing ngay cang gia tang. Viet Nam, mot nUdc dang phat trien, ciing khong n^m ngoai mo'i nguy hiem nay. Trong no lUc tim kie'm nhuEng che pham c6 tac dung ngan ngiifa va phong cho'ng ung thU, cac nha khoa hoc ciing quan tam den nhiing che pham cd kha nang tang cUdng siic khoe va siic cho'ng chiu cho ngUdi benh. Nam Linh chi la mot trong nhiing che pham do.

Linh chi tijf lau da dUdc biet den nhu mot loai "thSn dUdc" chijfa "bach benh". T\i trUdc den nay, da cd rat nhieu nghien cdu chi ra kha nang chuta tri mot so benh cua nam Linh chi, trong do cd benh ung thU. Chung toi Cling tien hanh mot so' nghien cxiu ve boat tinh khang u va kha nang ho trd dieu tri benh cua loai nam nay. Nhiing ket qua dUdc neu sau day chi de cap den anh hudng cua nam Linh chi len trong lUdng va thdi gian so'ng them cua chuot nh^t trang Swiss mang u bang sarcoma 180.

Cac nghien cijfu thUc nghiem dUdc tien hanh tren chuot bach tai phong thUc nghiem thugc Bo mon Te bao - Mo phoi va Ly sinh, Khoa Sinh hoc, TrUdng dai hoc Khoa hoc Tu nhien, Dai hoc Quoc gia Ha Noi.

215

NGUYEN LIEU VA PHUONG PHAP

Dong vat thi nghiem: chiing toi s\i dung chuot nh^t dUc trang,

dong Swiss, trong lUdng trung binh 20gam, tuoi trung binh t\i 7-8

tu^n, do Vien Ve sinh Dich t l Trung Udng cung cap.

Nam Linh chi (Ganoderma lucidum): chung toi sil dung san ph^m

da nghien nhd cua qua the va sinh kho'i sdi nam dang hot kho do cong

ty TNHH Nam Linh chi cung cap.

Tebdo ung thU sarcoma 180: Dong tebao ung thU sarcoma 180 la

dang u bang ac tinh thUc nghiem do Nhdm Ung thU thUc nghiem, Bp

mon Te bao-M6 phoi va Ly sinh, khoa Sinh hoc, trUdng Dai hoc Khoa

hoc Tu nhien cung cap.

Bo t r i t h i n g i e m

Thi nghiem 1: Anh hUdng cua thanh phan polysaccharide EFI

chiet tCf qua the va sinh kho'i sdi nam Linh chi nudi t rong len trong

lUdng chuot. (EFI la polysaccharide chiet trong dung moi la nUdc

va con).

Trong thi nghiem nay chung toi s\i dung 45 chuot, chia lam ba 16;

16 doi chiing (DC), 16 thi nghiem vdi EFI chiet tijf qua the (TNQ) va 16

thi nghiem vdi EFI chiet tu: sinh khoi sdi nam (TNS). 0 16 d6i chiing

tii ngay thtt ba sau cay truyen te bao ung thU (TBUT), chuot dUdc cho

uo'ng nUdc cat bang xdranh 1ml c6 gan kim tiem ddu tron, xong thang

vao da day. Cho u6'ng nam ngay lien tiep (moi ngay mot l§n:

0,2ml/con). Ngay thii tam nghi. Tiep tuc cho uo'ng tdi ngay thii 16 vdi

lieu lUdng nhu tren. h 16 thi nghiem, ciing vdi each cho ud'ng noi tren

nhUng thay nUdc cat b^ng dung dich che pham EFI vdi lieu

30mg/0,2ml/l Ian x 10 Ian (sau 48 gid cay truyen), tUdng dUdng tong

lieu 300mg/lchuot. Tien hanh can chuot each nhat (hai ngay mot Ian,

ke tu: ngay b^t dau thi nghiem). Den ngay thii 16 ket thiic thi nghiem.

216

Thi nghiem 2: Xac dinh thdi so'ng them cua chuot n h i t tr^ng Swiss mang u bang sarcoma 180 dUdc dieu tri bang polysaccharide EFI chiet tii qua the va sinh khd'i sdi nam Linh chi nu6i trong.

Cach b6'tri thi nghiem cung tUdng tu nhU thi nghiem I. Tuy nhien de'n ngay thu: 16 khong ket thuc thi nghiem ma tie'p tuc theo doi thdi gian sd'ng, che't cua chuot.

Cac phifcyng p h a p

- Chung toi sii dung phUdng phap chiet n i t polysaccharide theo

Takashi Mizuno (1985) [7,8].

- PhUdng phap cay truyen te bao ung thu [3,4].

Dung cu, hoa chat cay truyen: Bdm tiem 1ml, bong, con, dung dich sinh ly c6 pH=7,2 hoac dung dich muoi sinh ly 0,9%. Diing kim tiem loai 18 G trich bang b 6 bung chuot da giii gid'ng tijf 6 den 10 ngay roi pha loang b§ng dung dich sinh ly sao cho dich chtia te bao c6 mat do la 5.106 te bao/ml. Sau do tiem 0,2 ml vao xoang bung moi chuot, tUdng dUdng 106 TBUT/ con. Sau khi cay truyen, chuot dUdc cham soc can than va theo doi sU phat trien u bang cua chiing.

- PhUdng phap xac dinh thdi gian so'ng keo dai them theo Norio

Nagamoto [5]

ILS(%) = ( Ts/Cs - 1) X 100%

ILS -Increase in Life Span (Su gia tang thdi gian so'ng them). Ts: Thdi gian song trung binh ciia chuot 16 d dieu tri. Cs: Thdi gian so'ng trung binh cua chuot 16 do'i chiing

K £ T QUA VA THAO LU^N

1. A n h hvTorng c u a p o l y s a c c h a r i d e l e n t r o n g IvfoTng c h u o t

thi nghiem

Ket qua ve sU thay doi trong lUdng chuot d 16 DC va hai 16 TNQ,

TNS cd u6'ng che pham EFI vdi lieu 30mg/con/lan dUdc trinh bay trong

bang 1 va hinh 1.

217

Bang 1. Trong Ivftfng t rung binh cua chuot d cac 16 trong thofi gian thi nghiem

Stt Ian can

1

2

3

4

5

6

7

8

9

Ngay

0

2

4

6

8

10

12

14

16

DC(g)

20,0

22,0

23,0

25,0

27,0

27,5

28,0

26,3

24,4

TNQ (g)

20,0

22,0

26,0

27,0

28,0

28.0

30,0

29,0

29,0

TNS (g)

20,0

21,0

23,0

25,0

27,0

28,0

28,0

29,5

31,6

35

30

25

20

15

10

5

0

Trong luang (g)

—A-

DC TNQ

TNS

Thcri gian (ng^y)

0 8 10 12 14 16

Hinh 1: Do thi thay doi trong lUtfng cua chuot 16 d DC va hai 16 TNQ & TNS

Qua do thi tang trUdng chung tdi thay, trong ludng chuot d ca ba

16 DC, TNQ va TNS deu c6 xu hudng tang len d tuan d^u thi nghiem.

Rieng 16 TNQ, toe do tang trung binh c6 nhanh hdn so vdi 16 DC va 16

TNS. 6 tuan thii hai, sU thay doi trong lUdng d ca ba 16 la khong ro

218

ret. Sang tuan thii ba, trong lUdng trung binh cua chuot d 16 DC b i t

dau giam xu6'ng, cd the do tebao ung thU tang sinh manh lam can kiet

chat dinh dUdng cung cap cho cac te' bao lanh cua cd the. Day ciing la

hien tUdng thUdng thay d nhiing cd the mdc benh ung thU.

L6 TNQ cung c6 hien tUdng giam trong lUdng, tuy nhien to'c do

giam it hdn. L6 TNS lai cd xu hUdng tang trong lUdng vao nhuEng ngay

cuoi tuan thii ba. Day cd the la do ke't qua lam tang cUdng siic khoe do'i

vdi chuot nhdt t r i n g dUdc dieu tri b ing EFI. Mat khac, c6 the polysac­

charide chiet t\i sinh khd'i sdi nam Linh chi c6 tac dung utc che sU phat

trien cua te bao ung thU nen lUdng chat dinh dUdng cung cap cho cac

te bao lanh khac tang len d^n den sU tang trong lUdng cd the.

2. Anh hifcfng cua polysaccharide len thofi gian song keo

dai them cua chuot

Kha nang keo dai thdi gian sd'ng cua dong vat thi nghiem la mot

trong nhiing chi tieu quan trong de danh gia tac dung dieu tri ung thU

cua che pham. NhU da noi trong ph^n dd'i tUdng va phUdng phap

nghien ciiu, chiing t6i tien hanh chia 16 va theo doi sU song chet cua

chuot. Nhiing chuot che't deu dUdc giai phSu de kiem tra nguyen nhan

gay che't, de bie't ch§c ch^n 100% chuot che't la do ung thU.

Ket qua theo doi duac trinh bay trong bang 2, hinh 2.

Qua bang so' lieu tren ta thay, d hai 16 thi nghiem thdi gian s6'ng

keo dai them so vdi 16 doi chiing la 32,4% doi vdi 16 TNQ va 32,2% doi

vdi 16 TNS. Thdi gian so'ng trung binh cua chuot 16 doi chiing la 18,53

ngay (ke't qua nay phii hdp vdi nghien ciiu cua Nguyin Thi Quy vdi

thdi gian sd'ng trung binh cua chuot nh^t den thuan chung C57-BL6

mang u bang sarcoma 180 la 20.1 ngay). Thdi gian so'ng t r u n g

binh cua chuot ha i 16 thi nghiem TNQ - TNS keo dai hdn

khoang 6 ngay. Dae biet , moi 16 thi nghiem deu cd hai chuot

con sd'ng sot s au ngay thii 40.

219

Bang 2. Thdi gian song them ciia chuot nhat t r^ng d cac 16 DC, TNQ va TNS.

Stt chuot

1. 2. 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13.

T ILS (%)

Thdi gian song sot ciia chuot (ngay)

DC 13 13

14 14 15 15 15 17 17 19 21 23 25 26 31

18,53

0

TNQ 9 14

16 17

19 21 21 23 23 25

30 32 36 41 41

24,53

32,4

TNS 15 15 19 19 19 20 21 22

22 26 28 28 34 40 40

24,5 32,2

ngay 30 n

25 -

20 -

15 -

10 -

5 -

n -

24.5,

18.53

^M

-̂ ^̂ ^

^^k ^m>

U ~ 1

DC TNQ

3 24.5

H 1 1 H mm

\ TNS

L6 1

Hinh 2. Thdi gian so'ng t rung binh cua chuot 16 DC va hai 16 thi nghiem TNQ & TNS

220

Nhu vay, khi cho chuot bi ung thU uo'ng che pham chiet tii sinh kho'i sdi nam Linh chi khong nhiing c6 tac dung khang u (nhU da de cap trong mot nghien ciiu khac cua chiing toi) ma con cai thien thdi gian s6'ng them cua chuot theo chieu hUdng tot hdn. D6'i vdi che' ph^m chiet tijf qua the nam Linh chi, tuy chua the hien hieu lUc khang u (ciing theo nghien cutu tren), song no ciing c6 tac dung keo dai thdi gian song them tUdng dUdng vdi che'pham chiet tut sinh khd'i sdi nam. Dieu nay cd the la do che pham c6 tac dung tang cUdng siic khoe, do d6 tang kha nang cho'ng chiu ciia dong vat thi nghiem d in de'n keo dai thdi gian song. Hy vong d nhiing thi nghiem tac dong thuoc bang each tiem true tiep che' pham vao xoang bung, ke't qua se cao hdn.

KET LLIAN

Che pham polysaccharide chie't tOf qua the va sinh kho'i sdi nam Linh chi theo phudng phap cua Takashi va cong sU cd tac dung lam tang trong lUdng cua chuot thi nghiem mang u bang sarcoma 180. Dong thdi, ca hai che pham tren deu c6 kha nang keo dai thdi gian sd'ng them cua chuot thi nghiem len hdn 32% so vdi chuot do'i chiing.

TAI LIEU THAM KHAO

1. Nguyen Thi Chinh, Vii Thanh Cong (1999). Nghien ciJtu cong nghe san xuat sinh khoi vd mot so hoat tinh sinh hoc cua nam Linh Chi Ganoderma lucidum, Tap chi Sinh hoc. Tap 21, s6' 3: 132-136

2. D.K. Hossfeld, L.D. Sherman, R.R. Love, F.X. Bosch, 1991. Ung thu hoc Idm sdng (ban dich tieng Viet). NXB Y hoc. Ha noi.

3. N g u y i n Thi Quy (1993). Gay tao mo hinh u bang thitc ngiem vd thii tac dung phong vd chong ung thilcua mot so che pham tu nhien vd tong hop. Luan an Tien sy Sinh hoc, Trudng Dai hoc Tong hdp Ha noi.

4. T r i n C6ng Yen, 1975. Ciclul evolutiv al celulelor ascitice HR-

18 si influenta acestora asupra celulelor neurosecretoare din NSO

si NPV in conditii de tratament. Luan an Tien si Sinh hoc, Bucarest. 158pp (tieng Rumani).

221

5. Norio Nagamoto et al., 1978. Antitumor crocosmiflora, Planta medica.

6. Y. Quyan, Q. Yan. (1994). The antitumor effect of the G.luci-dum. Int. Sym. On Ganoderma Res, Beijing, China.

7. M. Takashi. (1996). A development of Antitumor Polysaccharides from Mushroom Fungi, FFI Journal 167: 69-85

8. M. Takashi. (1985). Fractionation and Characterization of Polysaccharide from Maitake, Grifla frondosa, Shizuoka, Japan.

SUMMARY

EFFECT OF POLYSACCHARIDE EXTRACTED FROM THE BODY AND MICELLIUM BIOMASS OF CULTURED GANODERMA LUCIDUM TO THE BODY WEIGHT AND THE INCREASE IN LIFE SPAN OF SWISS WHITE MICE CARRYING SARCOMA 180.

*Quy N.T., Nhung H.T.M, Duyen D.H., Hanoi National University.

We used 45 mice devided into three groups: the reference; the treated by EFI extracted from the fruit body and the those treated by EFI extracted from the micellium biomass of Ganoderma lucidum.

With total dose of 300mg / mouse (per.os, daily), the EFI of the micellium biomas and fruit body of Ganoderma lucidum increased the body weight of Swiss white mice carrying sarcoma 180. These products also to cause increase of the life span of the treated mice by 32% in comparison with the control mice (means 24.5 days and 18.53 days respectively).

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