Nghien cvcu YDHCT Viet Nam Sd40 - 2014
^ NGHIEN CUtJ TAG DUNG HA GLUCOSE MAU
CUA CAO LONG BOI M A U QUA LAU TREN C H U Q T N H A T BINH THUCJNG VA CHUOT NHAT DAI THAO D U Q N G T Y P 1
Studying the hypoglycemic action of extract B M Q L in normal mice and type 1-like diabetic mice
* Pham Thi Van Anh ', ** Nguyen Thi Thanh Ha ',
*** Nguyen Tr3n Thi Giang Hwong ' *** Tran Quoc Binh ^ ' Khoa YHCT Trudng Dgi hoc Y Hd Noi, ^ Binh viin Y hoc co truyen Trung uang Tom T5t:
Mac tiiu: Ddnh gid tdc dung ha glucose mdu cua cao long BMQL trin chuot nhdt binh thudng vd tren chuot nhdt DTD typ J gdy boi STZ. Phu&ngphdp nghi&n citu: Mo hinh: chuot binh thudng vd chuot DTD typ 1 gdy bdi STZDgng vdt nghien cuu- chugt nhdt trdng chiing Swiss. Thuoc nghien cuu- chuot binh thudng: gliclazid 80mg/kg the trong, BMQL 4,84g/kg, I9,36g/kg uong lien tuc 4 tudn; chuot DTD typ 1: STZ tiem mdng bung liiu 150mg/kg thi tronginsulin tiim mdng by.ng lUI/kg the trong, BMQL 4,84g/kg. 19,36g/kg the trong uong lien tuc trong 7 ngdy. Ket qud nghiin ciru:BMQL khdng co tdc diing hg glucose mdu tren chuot binh thuang sau 4 tudn uong thuoc va chuot DTD typ 1 sau 7 ngdy udng thu6c(p > 0,05). Kit luan: BMQL liiu 4.84g/kg vd 19.36g/kg khong CO tdc dung hg glucose mdu trin chuot binh thudng vd chuot DTD typ I.
Tir khoa: chuot nh5t dai thao dirtmg, bfii mSu, qua lau, BMQL, STZ, glucose mau Summary
STUDYING THE HYPOGLYCEMIC ACTION OF EXTRACT BMQL IN NORMAL MICE AND TYPE 1-LIKE DIABETIC MICE Object: to investigate the hypoglycemic action of extract BMQL in normal mice and STZ induced type 1-like diabetic mice. Methods: Model: normal mice and STZ induced type 1-like diabetic mice. Animal used: Swiss mice. Dose- normal mice: gliclazid 80mg/kg bw, BMQL 4,84g/kg, 19,36g/kg bw. oral route, during 4 weeks; type 1-like diabetic mice: STZ 150mg/kg bw i.p route, insulin lUI/kg bw. i.p route, BMQL 4,84g/kg. 19.36g/kg bw, oral route, during 7 days. Results: BMQL has no blood glucose - lowering effect in normal mice for 4 weeks treatment, and in type 1-like diabetic mice for 7 days treatment (p > 0.05) Conclusion:
extract BMQL has no effect of lowering blood glucose level at dose of4,84g/kg and 19.36g/kg daily in normal mice and type 1-like diabetic mice.
Key words: diabetic mice, bei mu, gua lou, BMQL, STZ, blood glucose
Dia chi lien he: Pham Thi Van Anh - Trudng Dai lipc Y Ha Npi Ngay nhSn bai: 12/01/2014 Dia chi email: [email protected] Ngay phan bien: 24/02/2014 Phan bien khoa hoc: PGS.TS. ViJ Nam Ngay dang bao- 24/3/2014
* TS.BS; ** ThS.BS; *** PGS.TS
Journal of research in viet nam traditional medicine and pharmacy No 40-2014
. DAT VAN DE
Dai thao ducmg (DTD) la benh ly rli loan chuyen hoa man tinh dac trung boi sir tang glucose mau do thieu insulin tuyet dli hay tucmg d6i. Cac thuoc y hgc hien dai dieu tri co hieu qua tit, nhung lai gay nhieu tac dung kh6ng mong mu6n va con nhieu ch6ng chi dinh, gia thanh cao [4]. Vi vay mgt trong nhung xu huang hien nay de dieu tri DTD la sir dung cay con thu6c CO nguin goc tu nhien, vira ban che dugc cac tac dung kh6ng mong muln lai thich hgp cho dieu tri keo dai.Bli mSu qua lau (BMQL) la bai thuoc c6 phuong CO tac dung thanh nhiet hoa dam. Cac vi thuic trong bai: Boi mlu, Thien boa phin. Cat canh, Tran bi.
Qua lau, Phuc linh khi diing rieng re da dugc bag cao co tac dung ha glucose mau tren thuc nghiem [1], [4].
Chiing t6i tiln hanh dl tai nghien cim nay vai mue tieu: Bdnh gid tdc dung hg glucose mdu ciia bdi thuoc BMQL tren chuot nhit binh thuang vd chuot nhdt DTD typ 1 gdy bai STZ.
II. DOI TirONG VA PHU'ONG PHAP NGHIEN ClJtJ
2.1. Doi tugng nghien cihi:
- Thuoc nghien ciru: Che pham thuic thu BMQL dang eao long san xuat tai Khoa Dugc, Benh vien Y hgc c l ttuyin Trung uong. C6ng thirc bai thuic nghien ciru gim 6 vi sau: Boi
mlu (2,5g), Qua lau (3g), Thien hoa phin (3g), Tran bi (3g), Cat canh (3g), Phuc linh (3g) (dugc gia giam khoi lugng tir bai thu6c B6i mau qua lau trich trong cuon Y hgc tam ngg)
- Dgng vat nghien ciru;
Chugt nhit trang chiing Swiss, khoe manh, gi6ng due, trgng lugng trung binh 25± 2g do Vien Ve sinh dich te trung uong cung cap. Dgng vat thuc nghiem dugc nu6i 7 ngiiy truoc khi nghien ciru va trong su6t thai gian nghien cuu bang thiic an chuan, nuac u6ng tu do.
2.2. Dung cu may moc va hoa chat nghien ciru
- Hoa chat phuc vu nghien cira + Streptozotocin (STZ) lg lg cua hang Sigma-Aldrich, Singapore
+ Diamicron (gliclazid) vien nen 30mg do hang Servier (France) san xuat.
+ Insulin lg 10ml, 40UI/ml, biet dugc Scilin R cua c6ng ty Bioton SA, Ba Lan.
+ Kit dinh lugng glucose On Call Plus cua hang ACON Biotech, My
+ Dung dich dem Cittat pH 4.5 - May moc phuc vu nghien cira + May thir duong huyet On Call EZII cua hang ACON Biotech, My
2.3. Phugng phap nghien cihi
Nghien cim YDHCT Viet Nam So 40 - 2014 2.3.1. Nghien ciiu tdc dung hff
glucose mdu tren chuot nhdt binh thir&ng
Chugt nhit tting dugc chia lam 4 16, m6i 16 10 con.
L6 1: Lo chimg (u6ng nuac cat) L6 2; U6ng gliclazid lieu 80 mg/kg.
Lo 3: Uong cao long BMQL lieu 4,84 g/kg (Tuang duang lieu lam sang)
L6 4: U6ng cao long BMQL lieu 19,36 g/kg (Gap 4 lan lieu lam sang)
Chugt dugc u6ng nuoc eat hoac thu6c thir lien tuc trong 4 tuan vao cac bu6i sang. Truoc khi lay mau lam xet nghiem chugt dugc nhin doi qua dem, lay mau du6i chugt, tien hanh dinh lugng duong mau tai cac thoi dilm t„
(chua u6ng thuoc), ti (sau u6ng thu6c thu 1 tuan), t2 (sau u6ng thu6c thur 2 tuan), t3 (sau uong thu6c thir 4 tuan).
2.3.2. Nghien cii'u tdc dung hg glucose mdu tren chugt nhdt BTB typ 1 gdy bai STZ
Chugt dugc chia thanh 5 16, moi 16 15 con (16 1 can 10 con)
L6 1: tiem dung moi + uong nuoc cit0,2ml/10g
L6 2: tiem STZ 150mg/kg + uing nuoc cat 0,2ml/lOg
L6 3: tiem STZ 150mg/kg + tiem mang bung insulin lieu lUMcg
L6 4: tiem STZ 150mg/kg + Uong cao long BMQL liiu 4,84 g/kg
L6 5: tiem STZ 150mg/kg + U6ng cao long BMQL liiu 19,36 g/kg
Tit ca chugt a cac 16 dugc djnh lugng glucose mau lan 1 sau khi nhin doi qua dem (To). Cac 16 chugt tir 16 2- 5 dugc tiem STZ liiu 150mg/kg. Sau 72 gig tiem STZ, dinh lugng glucose mau lan 2 (Tyih), chgn cac chugt hi dai thao duong co mue glucose khi doi hen lOmmol/1 dugc tham gia nghien curu. Cac 16 chugt dugc uIng nuoc cit hoac thuoc thu hoac tiem insulin trong 7 ngay sau khi gay m6 hinh, djnh lugng glucose mau luc doi tai thoi diem sau uong thuoc thu 7 ngay (Tc).
2.4. Xiir ly so li^u: S l lieu dugc nhap va xu ly bang phucmg phap va thuat toan thong ke y sinh hgc tten phan mem Excel 2007. So lieu dugc bieu diln dudi dang X ± SD. Kiem dinh cac gia tti bang t-test Student hoac test tnrac-sau (Avant - Apres).
Su khac biet co y nghIa th6ng ke khi p<0,05. Chu thich:": Khdc biet so vdi chirng sinh hoc v&i p<0,05, vd***: p<0,OOI
HI. KET QUA NGHIEN CiTlJ 3.1. Tac dung ha glucose mau tren chugt nhit binh thuong
Journal of research in viet nam traditional medicine and pharmacy No 40 - 2014
Bang 1. Nong do glucose mau ciia chuot nhit binh thugng sau 4 tuan uong thuoc
Lo chugt L6 1: Chung sinh hgc L6 2: Gliclazid 80mg/kg
L6 3: BMQL 4,84g/kg
L6 4:BMQL 19,36g/kg
Nong do glucose mau mmol/1 (X ± SD) n = 10 T„
5,53 ±1,02 5,24 ±1,54
5,65 ± 0,95
5,27 ± 0,98 T, 5,34 ±1,02 3,70 ±1,78 p2-l<0,0S 5,41 ±1,39 p3-l>0,05 5,28 ±0,71 p4-l>0,05
T j
5,50 ±0,61 2,90 ±1,29 p2-l<0,001
5,61 ±1,51 p3-l>0,05 6,04 ± 0,78 p4-l>0,05
T3 5,92 ±1,25 4,13 ±0,49 p2-l<0,001 5,38 ± 0,90 p3-l>0,05 6,34 ± 0,57 p4-l>0,05 K6t qua bang 1 cho thiy: Cao long BMQL ca 2 lieu uong lien tuc sau 4 tuan khong lam giam nong dp glucose mau so voi 16 chung (p > 0,05). Gliclazid lieu 80mg/kg CO tac dung lam giam nong dp glucose mau ro ret so voi nhom chOmg (p< 0,001).
3.2. Tac dung ha glucose mau tren chuot nhat DTD typ 1
Bieu do 1. Can nang cua chuot nh^t DTD typ 1 qua cac then diem
Nghien cUu YDHCT Viet Nam So 40 - 2014 Nhan xet: Can nang ciia cac 16 chugt dugc tiem STZ sau 72 gig diu giam co y nghia thing ke so vai 16 chirng (p < 0,05). Sau 1 tain uIng cao long BMQL, a ca 2 16 uIng thuic thil liiu 4,84g/kg va 19,36g/kg, 16 tiem mang bung msulin va 16 mo hmh, chugt kh6ng tang can sg vai thcri dilm sau tiem STZ 72 gia va van thap hon ro ret so vai 16 chiing sinh hgc (p< 0,001). Khong co sir khac biet giira cac 16 chugt DTD typ 1 (p > 0,05).
Bang 2. Anh hugng eiia cao long BMQL len nong dg glucose mau cua chuot nhit tring DTD typ 1
Lo chugt Lo 1: Chimg sinh hgc Lo 2: Mo hinh
% giam glucose mau ciia Tc so vol T72h Lo 3:Insulin lUI/kg
% giam glucose mau cua Tc so voi Tj2h Lo 4: BMQL 4,84g/kg
% giam glucose mau cua Tc so vol T72h L6 5:BMQL19,36g/kg
% giam glucose mau cua Tc so voi Tjzh
Nong do glucose mau mmol/1 (X ± SD) T„
6,28 ± 0,87 6,37 ± 0,80
6,86 ±1,03
6,52 ± 0,90
6,08 ± 0,70
T72h
6,16 ±0,65 15,97 ±4,52 p2-l<0,001
15,07 ±4,96 p3-l<0,001
15,67 ±4,22 p4-l<0,001
16,06 ± 5,89 p5-l<0.001
Tc 5,41 ±0,78 10,81 ±3,56 p2-l<0,001
32,31 3,44 ±0,27 p3-2<0,001 77,17 9,15 ±3,01 p4-2>0,05
41,61 8,80 ± 3,65 p5-2>0,05
45,21 Ket qua bang 2 cho thay: O 16
chiimg sinh hgc, nong do glucose khong thay d6i qua cac thoi dilm. d cac 16 chugt tiem STZ, sau 72 gio, nong do glucose mau tang cao ro ret so
voi 16 chiing sinh hgc (p<0,001). Sau 1 taan uong thu6c, cao long BMQL ca 2 lieu CO xu huomg lam giam nong do glucose mau so vai 16 m6 hinh, nhung sir khac biet kh6ng co y nghia thing
Journal of research in viet nam traditional medicine and pharmacy No 40 - 2014
ke (p>0,05). Lo u6ng BMQL liiu 19,36 g dirpc lieu/kg lam giam 45,21%
nong dp glucose mau, cao hcfn 16 u6ng BMQL lieu 4,84 g duac lieu/kg la 41,61%. Insulin lieu lUI/kg co tac dung lam giam nong dp glucose mau ro ret so vcri 16 mo hinh (p<0,001), voi miic giam la 77,17%. O 16 mo hinh, miic giam glucose mau cung dat 32,31%.
VL BAN LUAN
4.L Ve tac dung ha glucose mau tren chuot nhat binh thirong
Chung toi tien hanh danh gia anh huofng cita bai thu6c BMQL len chupt nhat trang binh thuang trong thai gian 4 tuan, so sanh vai thu6c chiing duang la gliclazid. Gliclazid 80mg/kg lam giam glucose mau ro ret of ca 3 thai diem xet nghiem. Cao long BMQL a lieu 4,84g/kg va 19,36g/kg khong gay ha glucose mau a ca 3 thai dilm, nong dp glucose mau khac biet khong co y nghTa so voi truac khi nghien ciiu va so voi 16 chiing (p>0,05). Tren dpng vat binh thuong luon co ca che can b ^ g giGa cac yeu t6 gay ha glucose mau (insulin, incretin) va cac yeu to lam tang glucose mau (adrenalin, glucagon,...), chi khi cac tac nhan ben ngoai pha v5 duac he th6ng can bang nay moi lam cho nong dp glucose mau
thay doi ro ret [4], 6 day co the giai thich bang 2 ly do: (I)v6i lieu 4,84g/kg va 19,36g/kg, cao long BMQL chua dii manh de co the gay ha glucose mau, (2)thai diem xet nghiem glucose mau khong phai la thoi diem thuoc CO tac dung manh, tham chi khi do thu6c da het tac dung.Hien chua c6 nhieu nghien ciiu ve tac dung ha glucose mau cua cac dupc lieu trong bai thuoc, nghien ciiu ciia Hikino co n6i den tac dung ha glucose mau tren chupt nhat binh thuang cua 5 loai trichosan A, B, C, D, E tach chiet tu Thien hoa phan, tuy nhien chua c6 so lieu cu the [6], [7].
4.1. Ve tac dung ha glucose mau tren chuot nhat DTD typ 1
STZ la mpt thuoc chong ung thu, dupc sii dung de gay DTD thuc nghiem, co che la pha hiiy chon Ipc te bao p tuyen tuy, miic dp pha huy tuy thupc vao lieu lupng su dung, tren chupt nhat tu 100 - 200mg/kg, tren chupt cong tir 35 - 65mg/T;g [5]. Trong nghien ciiu nay, chiing t6i chpn lieu STZ 150mg/kg, lieu nay cung duoc tac gia trong va ngoai nuac sii dung de gay DTD typ 1[2], [3]. Bieu d6 1 cho thay sir giam trpng lupng chupt a thoi diem sau tiem STZ 72 gio va sau 1 tuan uong thu6c, thap hon ro ret so voi
NghUn ciru YDHCT Viet Nam So 40 - 2014
16 chiing sinh hgc (p<0,001).Ket qua nay tugng tu nghien ciru ciia mgt s6 tac gia tren the gioi, tten ca chugt nhat va chugt c6ng [6]. Can nang ciia chugt tiem STZ giam co the giai thich do insulin giam manh, glucose khong dugc van chuyen vao te bao, ca the mat ngu6n nang lugng cung cap tir glucose nen phai thay the bang qua trinh giang h6a cac chat beo, chu yeu tir ca van. Bang 2 cho thay lieu STZ 150mg/kg gay dugc mire tang glucose huyet cao tren 15 mmol/1, tuong tu kit qua cua Tomoji F. la 16,7 mmol/1. He Lin T. la 17 mmol/1 [7].
De c6 tac dung tren DTD typ 1, thuoc dieu tri co the theo cac co chi sau: (1) c6 boat tinh tuang tu insulin tten chuyin boa glucid, (2) lam tang van chuyen glucose vao te bao kh6ng hen quan msulin, (3)tang thai glucose ra nuoc tiiu bang each ire chi SGLT2 a Ing than. Dilu tri DTD typ 1 cin mgt thuoc CO tac dung nhanh, manh nhu insulin de lam ha glucose mau, do biln chirng toan mau c6 the xay ra bit cir luc nao. Do vay, nghien curu nay kh6ng nham mue dich tim thuic co kha nang thay the insulin ma chi tham do CO chi cua cao long BMQL.
Insulin, hormon tuyen tuy, co vai tro
van chuyen glucose trong mau vao te bao, lam giam nong do glucose mau tai 77,17%. Q cac 16 u6ng cao long BMQL, glucose mau co xu hugng giam so vai 16 m6 hinh nhung kh6ng CO y nghia th6ng ke, (miic giam so vol thoi dilm T72h la 41,61 va 45,21%, co cao hem so voi 16 mo hinh 32,31%).
Co the cho rang thai gian 1 tuan uong thuoc chua du hoac lieu lugng chua dat miic de thuoc the hien tac dung.
BMQL la bai thuoc co phuang co tac dimg thanh nhiet h6a dam, nhuan phi chi khai (day la 1 trong cac phap dieu tti benh Tieu khat, co nhieu dac dilm gi6ng DTD). Nhieu nghien ciiii da bao eao tac dung ha glucose ciia cac dugc lieu trong bai thuoc [3], [7]. Satoko va cgng su bao cao hesperidin (trong Tran bi), lieu lOg/kg co tac dung giam glucose mau sau 4 tain dilu tri tten chugt cing DTD typ 1 gay bai STZ.
Hesperidin lam tang glucose kinase va giam glucose - 6 phosphatase, hesperidin khi vao ca thi dugc chuyen hoa thanh hesperetin, mgt bioflavonoid CO tac dung ha glucose mau qua eg chi tang sao ma adiponectin (protein tilt ra tir te bao tao ma, dong vai tro quan trgng trgng dilu hoa chuyen boa glucose va lipid, dilu chinh sir nhay
Journal of research In viet nam traditional medicine and pharmacy No 40 - 2014
cam insulin va can bang nang luang) va tang bilu Men PPAR-y- Nhu vay, neu thai gian nghien ciiu dai hon, tien hanh ducrc dinh luang cac yeu t6 tren thi CO the danh gia dupc chinh xac tac dvmg cua cao long BMQL len mo hinh chupt nhat DTD typ 1.
V. KET LUAN:
- Cao long BMQL ligu:
4,84g/ kg/ ngay va 19,36g/ kg/ ngay khong lam giam n6ng dp glucose mau a chupt nhat trang binh thuang sau 4 tuin dimg thu6c.
Cao long BMQL lieu:
4,84g/kg/ngay va 19,36g/kg/ngay co xu huong lam giam glucose mau tren chupt nhat trang DTD typ 1 gay boi STZ 150mg/kg.
TAI LIEU THAM KHAO 1. Nguyen Trong Thong (2010),
"Thuoc dieu chinh r6i loan glucose mau", Du(rc ly hpc tap 2, Nha xuat ban yhpcHaN6i,tr. 186-194.
2. Li W.L., Zheng H.C, Bukuru J. (2004), "Natural medicines used in the traditional Chinese medical system for therapy of diabetes mellitus", Journal of Ethnopharmacology 92, pp.
1 - 2 1 .
3. Xie W.. Du L., (2011)
"Diabetes is an inflammatory disease:
evidence from traditional Chinese medicines", Diabetes, Obesity and MetabolismniA), pp 289 - 301
4. Bo mon Mien dich-Sinh ly benh (2002), Sinh ly benh hoc, Truang Dai hpc Y Ha Npi, Nha xuit ban Y hoc, tr 59 - 71
5. Hikino H., Nakajima K., Sato S. (1989), "Isolation and hypoglycemic activity of trichosans A, B, C, D and E glycans of Trichosanthes kkilowh roots", Planta Medica 55, pp. 349 - 350.
6. Shahidul I., Rachel D.W.
(2012), "Animal Models in Diabetes Research: Experimentally Induced Rodent Models of Type 2 Diabetes ".Methods in Molecular 5/o/ogv955, p p l 6 1 - 1 7 4 .
7. Nguyen Ngoc Xuan {2QQA)J^ghien cim tdc dung hg duang huyet ciia Tho phuc linh (Smilax glabra roxb smilacaceae) tren site vat thuc nghiem, Luan an tien sy, tr. 54 - 75.
