Y Hoc TP. Ho Chi Minh * T^p 20 * So 6 * 2016 Nghien ciiu Y hpc
DANH GIA TAG DUNG DIEU TRI ROI LOAN UPID MAU CUA CAO LA SEN- HA THU 6 DO (FOLIUM NELUMBINIS -
RADIXPOLYGONI MULTIFLOm TREN THl/C NGHIEM
Huynh Thi My Hien*, Le Thi Lan Phuong*, Nguyen Thi Scm*, Nguyen Phucmg Dung*TOMTAT
Ma d&u: Roi loqn lipid mdu Id nguyin nhdn chinh gdy xo viea dgng mqch vd tdng tyli tie vong do benh ly tim mqch. Do dd dOu tri rdi loqn lipid mdu tro thdnh mdi quan tdm hdng dau. Ld Sen (LS) vd Hd thii d do (HTO) Id nhieng dugc liiu cd san Hong nuac vd da dugc kii hgp sie dung tren ldm sdng diiu Hi rdi hqn lipid mdu. Tuy nhiin chua co nghiin cieu chung minh hiiu qud dieu trf rdi loqn lipid mdu cua viec kei hgp Id Sen vdHdthiid dd.
Ve tdi ndy dugc thuc hiin nham muc dich cung ccqj cdc bang chimg khoa hgc cho viec ieng dung kei hgp Id Sm vd Hd thu d do Hong dieu tri rdi loqn lipid mdu.
Muc tieu: Khdo sdt dgc tinh cap ciui cao kei hgp Id Sen - Hd thii ddd(LS- HTO) ty li 2-1. Ddnh gid tdc dung dieu tri rot loan lipid mdu cua cao kii hgp Id Sen — Hd thii d do Hen thuc nghiem.
Doi tugng vd phuongphdp nghien cOu: Chudt nhat Hang, chung Swiss albino, Hgng lugng 20 ±2 g, miui tai Viin Pasteur TP. HCM. Nghien cieu thuc nghiem cd nhom chimg Hin md hinh rdi loqn lipid mdu ngoqi sinh (bang udng diolesterol 25 mg/kg).
Ke't qua: Dmax ciia cao Id Sen -Hdthitd^ld 8,99 g cao/kg chugt Trin mo hinh rm loqn lipid mdu ngoqi sinh, cao LS-HTO ty li 2-1 lieu LS 0,8 g dugc lieu/kg - HTO 0,4 g dugc lieu/kg gidm cholesterol todn phdn 40,5%, triglyceride 45,88%, LDL-C 43,16% . Cao LS-HTO lieu LS 0,4 g dugc lieu/kg - HTO 0,2 g dugc lieu/kg ldm gidm triglycerid 28,51%.
Kit lu4n: D<mx cao LS -HTO Id 8,99 g cao/kg (tuang duang 270,39 g dugc liiu khd/ngu&i 60 kg).Cao Id Sen - Hd thii 6 dded tdc dung diiu Hj rdi loqn lipid mdu Hen thuc nghiim.
Tit khoa: Ld sen, Hd thii d do, dieu tri rdi loqn lipid mdu.
ABSTRACT
EVALUATE DYSLIPIDEMIA TREATMENT EFFECTIVENESS OF THE EXTRACT COMBINED FROM FOUUM NELUMBINIS - RADIX POLYGONI MULTIFLORI IN MICE
Huynh Thi My Hien, Le I h i Lan Phuong, Nguyen Thi Son, Nguyen Phuong Dung
* Y Hoe TP. Ho Chi Minh * Vol. 20 - No 6 - 2016:181 -185 Background: Dyslipidemia is a major cause of atherosclerosis and increasing mortality due to cardiovascular disease. PrevmHon and Heatment of dyslipidemia has become an essential concern. Folium Nelumbinis (lotus leaf) and Radix Polygoni MulHflori havebeen available domestic herb. They have bem combined in dyslipidemia Heatmmt However, no study has demonstrated dyslipidemia preumHon effrd of Folium Nelumbinis and Polygonum Multifiorum combination. This research is conducted to provide scientific evidences for this combinaHon in treatment dyslipiikmia.
Aims of study: Survey the acute toxidty of aqueous exHact combined from Folium nelumbinis - Radix Polygoni multiflori (2-1 ratio). Evaluate dyslipidemia treatment effectiveness of Folium Nelumbinis - Radix
' Khoa Y Hpc Co Truyen - Dai hpc Y Duoc Thanh pho Ho Chi Minh
Tdc gid lien lac: ThS. BS. Huynh Thj My Hien DT. 01682089687 Email: [email protected]
N g h i d n Clhi Y hpc Y Hpc TP. H b Chi M i n h * T a p 20 * So 6 * 2016 Polygoni Multiflori exHact in hyperlipidemic mice.
Methods: Male and frmale Swiss albino mice, wdghing 20 ±2 g, were purchased at the Pastmr InsHtute in HCM City. Experimental study was conducted with control group on exogenous dyslipidemia model by oral adminisHaHon of cholesterol at a dose of 25 mg/kg frr 8 weeks.
Results: Dma of Folium Nelumbinis - Radix Polygoni Multiflori exHact is 11.818 g/kg. On the model of exogenous dyslipidemia, Folium Nelumbinis - Radix Polygoni Multiflori combinaHon (Folium Nelumbinis 0 8 g/kg - Radix Polygoni MulHflori 0.4 g/kg) reduced 40.5% total cholesterol, 45.88% triglyceride, and 43.16%
LDL-C. The combination of Folium Nelurribinis 0.4 g/kg - Radix Polygoni MulHflori 0.2 g/kg decreased 28.51%
triglyceride.
Conclusion: Dma of Folium Nelumbinis - Radix Polygoni MulHflori extract is 11.818 g/kg (equal to 270,39 g herb/60 kg person). Folium Nelumbinis - Radix Polygoni MulHflori exHad has hypolipidemic effed on cholesterol-induced exogenous hyperlipidemia in mice.
Key words: Folium Nelumbinis, Radix Polygoni Multiflori, dyslipidemia Heatment effectiveness.
D A T V A N D E
Roi loan lipid mau la yeii to thuan Ipi gay nhieu benh ly tim mgch do xa vua dgng mach (XVDM). Theo thong ke, ty le tii vong 6 benh nhan XVDM la 45%, trong do berth mach vanh chiem 32% va tai bien mach mau nao chiem 18%'^'. Do do, dieu tri roi loan lipid mau tro thanh mpt trong nhiing moi quan tam hang dau trong cham soc siic khoe cua nhieu quoc gia.
Nhieu bai thuoc va vi thuoc tu thien nhien da dupe chiing minh la co tac dung dieu hoa lipid mau nhu Ngim tat. Qua lau. Ha thii 6 do.
Ban ha, la Se'^' ^""' i"^**"*"™ w""^ not found.) Xrong thuc te lam sang, nhieu thay thuoc da ket hpp la Sen va Ha thii 6 do trong dieu tri roi loan lipid mau, tuy nhien ehua co nghien ciiu chiing minh hieu qua dieu trj roi loan Upid mau ciia viec ket hpp 2 dupe Ueu tren. De tai nay dupe thuc hien nham tim hieu tac dung dieu tri roi l o ^ Upid mau khi phoi hop la Sen va Ha thii 6 do tren mo hinh thue nghiem.
Muc tieu ciia nghien ciiu la khao sat dpc tinh cap eiia cao k^t hpp la Sen - Ha thii 6 do ty le 2-1 va danh gia tac dyng dieu tri roi loan Upid mau hen chupt nhit tr^ng ciia cao la Sen - Ha thu 6 do tren mo hinh chupt nhSt trSng roi loan Upid mau ngoai sinh (uong cholesterol).
DOIlUONG-PHl/ONGPHAPNGHIENCLrtj Nguyen lieu n g ^ e n o i u
Cao la Sen (LS), cao Ha thii 6 do (HID) dupe bao che bang each sac 2 lan voi nuac, Ipc didi chiet dem eo de dupe cao. Cao LS eo hieu suat chiet 16,34%, dp aim 12,94%, dp tro 5,31%. Cao HTO CO hieu suat chiet 16,99%, dp am 15,54%, do tro 4,15%. Djnh tinh bang sac ky lop mong cao LS, cao HTO cho cac vet co mau sac va Rf tuang tu voi mau doi chieii LS, HTO.
P h u o n g tien
Hoa chat: Cholesterol, acid choUc (Sigma).
Thuoc doi chieii: Atorvastatin lOmg (Stada - Viet Nam).
D p n g vat n g h i e n curu
Chupt nhat trang Swiss albino, khoe mgnh, dong deu ve gioi tinh, 6-8 tuan tuoi, trpng lupng 20 + 2 g, cung cap boi Vien Pasteur TP. Ho Chi Minh. Trong suot qua triiih thii nghiem, chupt dupe nuoi trong dinh duang on dinh.
P h u o n g p h a p n g h i e n cmi
Khao sat dpc tinh cap ciia cao la Sen - Ha thii 6 do'i^'.
Cho chupt nhin an 12 gia truoc khi thii nghiem, uong nuoe tu do. Dua thuoc duong uong bang kim d^u tu mpt Ian duy nhat vai the tieh 0,2 ml cao/lO g the hpng.
182
Y Hoc TP. Hb Chi Minh * Tap 20 * So 6 * 2016 N g h i e n cuu Y hpc
Giai doan tham do (4 chup1/l6): Khoi dau tix
^e\i cao nhat co the bom duoc qua kim cho uong. Xae djnh LDo (Ueu toi da khong gay chet) va LDiDo (lieu toi thieu gay chet 100%).
Giai doan xac dinh (20 chu6t /to): Chupt dupe chia 16, moi 16 duac cho uong thuoc thii nghiem mpt Ueu suy nhat trong khoang LDo va LDioo chia theo cap so nhan.
Danh gia ket qua:Theo doi cac bieu hien ve hanh vi va van dpng ciia chupt trong vong 72 gio sau khi dimg thuoc. Ghi nhan so chupt chet trong timg 16. Xac dinh Dnux hoac LDso theo phuong phap Karber-Behrens (neu eo) de lam ca sa tinh Ueu cho cae thii nghiem dupe ly.
Danh gia tae dung dieu tri roi loan Upid mau ciia cao ket hpp la Sen - Ha thii 6 do tren m6 hinh gay roi loan Upid mau bang cholesteroic*?).
Chupt thii nghiem sau khi nuoi on dinh 1 tuan dupe chia lam 216:
- L6 chiing (n=12): Uong nuac cat.
- Lo cholesterol (n=36): Uohg cholesterol 25 mg/kg trong dau thuc vat.
Dung dich cholesterol trong dau hoac nuoc cat dupe cho UOTig vao 8 - 9 gia sang m6i ngay voi the tieh 0,1 mI/10 g trong 6 tuan. Sau do, 6 chupt thuoc 16 chiing va 6 ehupt thupe 16 cholesterol dupe gay me va lay mau true Hep tii tim chupt de dinh luong cholesterol TP, TG, HDL-C va LDL-C- So chu6t con lai dupe chia ihanh 516, m6i 16 6 chupt:
- Lo chiing: Uong nuac cat.
- L6 benh: Uong cholesterol 25 mg/kg trong dau ftiuc vat va nuac cat.
L6 atorvastatin (16 doi chiing): Uong cholesterol 25 mg/kg trong dau thuc vat va atorvastatin 10 mg/kg.
- Lo LS-HTO (1): Uong cholesterol 25 mg/kg trong dau thuc vat va eao la Sen - Ha thii 6 do vol lieu LS 0,8 g dupe Ueu/kg va HTO 0,4 g dupe Ueu/kg.
- Lo LS-HTO (2): Uraig cholesterol 25 mg/kg trong dSu thuc vat va cao la Sen - Ha thii 6 do
voi Keu LS 0,4 g dupe Ueu/kg va HTO 0,2 g duoe Ueu/kg.
Dung dich cholesterol trong dau hoac nuac cat (doi voi 16 chung) dupe cho uong vao 8 - 9 gia sang moi ngay, eao LS - HTO hoac atorvastatin eho uong vao 3 -4 gia chieu m6i ngay voi the tieh 0,1 ml/10 g trong 2 tuan.
Sau 8 tuan thii nghi|m, tien hanh gay me va lay mau true tiep tir tim chupt de dinh lupng cholesterol toan phan, triglycerid, HDL- C va LDL-C bang phuang phap do quang (thuoc thu dinh lupng Upid mau ciia cong ty Maxmat - Phap).
Phuong phap x u ly thbng ke sb lieu thuc
nghifm
Cac dii Ueu dupe trinh bay duoi dang Mean
± SEM. Xii ly thong ke bang phan mem SPSS phien ban 16.0. Phmmg phap Ihong ke dupe sii dung la phep kiem Student cho 2 day so Ueu dpc lap. So sanh su khac nhau giiia cac nhom co tac dung dieu tri roi loan Upid mau bang phep kiem ANOVA mpt chieu, mpt yeii to. Su khac nhau dupe xem la co y nghia khi gia tri p < 0,05.
KET QUA VA BAN LUAN Dpc tinh cap
Trong 2 tuan quan sat, khong gju nhan dupe bat ky dau hieu bat thuong nao tren chupt thu nghiem. Tat ca chugt deu an uong va hoat dpng binh thuong, kh6ng eo chupt ehet. Do do khong tim dupe LD50.
Dn.a> ciia cao LS - HTO la la 8,99 g cao/kg, tuong duang 270,39 g dupe Ueu kho/nguai 60kg, gap 34 Ian so voi lieu su dung tren nguai. Dieu nay eho phep nhan djnh rang ket hpp cao la Sen va cao Ha thii 6 do sii dung trong thii nghiem an toan doi voi dpng vat thii nghiem, phii hpp vai thuc te sii dung ket hpp la Sen va Ha thu 6 do chua ghi nhlin cac tac dung bat Ipi tren benh nhan.
De tai chua co dieu kien khao sat tac dung phu ciia cao la Sen - Ha ftiii 6 do khi sii dung dai ngay. Thoi gian dimg thuoc dieu tri roi loan Upid
N g h i e n c u u Y hpc Y Hpc TP. Hb Chi M i n h * T?p 20 * S b 6 * 2016
mau thuang keo dai. Vi the, can tiep tuc tioi hanh nghien ciiu dpc tinh ban truong dien ciia cao la Sen - Ha thii 6 do de lam co so cho cac buoc trien khai lam sang sau nay.
Danh gia tac dung dieu tri roi loan Upid mau ciia cao ket hpp la Sen - Ha thii 6 tren m6 Mnh gay roi loan Upid mau bang cholesterol
So vai 16 chiing, chupt uong cholesterol 25 mg/kg trong dau thuc vat co nong dp cholesterol TP tang gap 1,5 lan (p < 0,05), TG tang gap 1,36 1 ^ (p < 0,05) va LDL-C tang gap 10,35 lan (p < 0,01). Rieng nong dp HDL-C khong CO su khac biet giiia 2 lo.Nhu vay, sau 6 Bdngl. Ndng do cholesterol todn phan, triglycerid Hong mdu ngoqi sinh bdng cholesterol
tuan uong cholesterol 25 mg/kg, da tao dupe tmh trang tang cholesterol TP, TG, LDL-C tr^n chupt nhat trang co y nghia thong ke.
Bdng 1. Ndng dd lipid mdu ciia chudt nhat Hang sau 6 tudn gdy binh bang cholesterol (mg/dl)
Cac thong s6 lipid m i u Cholesteroi TP
(mg/dl) TG (mg/dl) LDL-C (mg/dl) HDL-C (mg/dl)
Lo chimg (n=6}
112,33±16.56 122,92 ±17,09 5,75*2,48 81,99±17,09
Ld cholesterol (n=6) 170,43 ±50,72"
166,66 ±16,64' 59,64 ±36,53 "
83,43 ±16,91 Ghi chu: Khde biet cd y nghia thdng ki so vai nham dtung (*: p< 0,05, **:p< 0,01).
mdu chudt nhat sau 8 tudn g&y md hinh rdi loqn lipid L6{n = 6)
Chung Benh Atorvastatin LS-KTO (1) LS-HTO (2)
Thudc ttiLF, Li§u
Nirticcat
Cholesterol {25 mg^g) Cholesterol (25 mg/kg) Atorvastatin (10 mg/kg) Cholesterol (25 mg^g) LS(0,8g)-HTO(0,4g) Cholestertrf (25 mgftg) LS(0.4g)-HTO(0,2g)
ChoIesterviTP (mg/dl) 122,14 ±23,74 265,58 ±62,93 175.88 ±106,13**
155,37 ±94,58"*
238.04 ±42,02 "*•
TG (mg/dl) 132,98 ±19,70 133,99 ±22,61 101.52 ±51.20"*
72,52 ±44,23"**
95,78 ±9,15"^
Ghi chu: Khde biit cd y nghia thmg ke so vai nhdm chieng (*: p <
nghia thong ki so v&i nhdm binh (9: p < 0,05, ##; p< 0,01).
0,05, **; p < 0,01, ***: p < 0,001); Khac biet cd y
Nhan xet: So voi chiing, 16 b | n h co nbng dp cholesterol TP tang 2,17 lan (p < 0,001).
Chiing to m6 hinh roi loan chuyen hoa cholesterol ngoai sinh van duy tri den tuan thii 8 (sau khi da ngung uong cholesterol 2 hian) tren chupt thii nghiem.016 atorvastatin, nong dp cholesterol TP giam 33,77%;TG giam 24,22% so vpi 16 benh (p < 0,05), phii hpp voi hieu qua dieu hoa roi logn Upid mau cua statin nay<«. So vai 16 benh, ehupt trong 16 LS- HTO
Bang 3. Nong dd HDL-C vd LDL-C Hong mdu chudt nhdt sau 8 tiian gdy md hinh roi loan lipid mdu ngoqi sinh bang cholesterol
(1) CO nbng dp cholesterol TP giam 41,5% (p <
0,05), TG gi^m 45,88% (p < 0,01). L6 LS - HTO (2) CO nbng dp TG giam 28,51% (p < 0,01).
Nhu vay, ket hpp LS - HTO voi h^u LS 0,8g dupe Ueu/kg - HTO 0,4 g dupe U?u/kg eo tac dung lam ha cholesterol TP va TG; trong khi U ^ thap hon (LS 0,4g dupe USu/kg - HTO 0,2 g dupe Ueu/kg) chi lam ha TG tren ehupt nhat trang tang cholesterol ngoai sinh.
L6(n = 6) ChiJTig
Bgnh Atorvastatin LS-HTO (1) LS-HTO (2)
Thu6c thir, lieu Ntroc cM Cholesterol (25 mg/kg) Cholesterol (25 mg/kg) Atorvastatin (10 mg/kg) Cholesterol (25 mg/kg) LS(0,8g)-HTO(0,4q) Cholesterol (25 mg/kg) LS(0,4g)-HTO(0,2g)
HDL-C (mg/d)) 75,15 ±20,48 75,90 ±17,14 57,90 ±31,65 48,56 ±30,62 60,70 ±10,17
LDL-C (mg/dl) 19,36 ±13,69 164,10±61,48 102,11 ±70,77"*
93,28 ± 56,98~*
158,15 ±43,44"'
184
Y Hpc TP. Hb Chi Minh * Tap 20 * Sb 6 * 2016 Nghien cuu Y hpc
Qii chu: Khde biet co y nghm thdng ke so v&i nhom chiing nhdm binh (tt: p < 0,05)
Nhan xet. So vol 16 benh, nong dp LDL-C ciia 16 LS - HTO (1) giam 42,2%(p < 0,05),tuang tu 16 ubng atorvastatin. Rieng nbng dp LDL-C ciia 16 LS - HTO (2) kh6ng co sir khae biet thong ke so voi 16 benh.Nhu vay, cao LS -HTO co tac dung ha LDL-C 6 lieu LS 0,8 g dupe Ueu/kg - HTO 0,4 g dupe Ueu/kg
Ket qua bang 2 va being 3 eho thay cao phoi hpp LS-HTO a Ueu LS 0,8 g/kg va HTO 0,4 g/kg CO tac dung dieu ehinh roi lo?n Upid mau tr§n chupt nhat tang Upid mau ngoai sinh sau 2 tuan dKU trj, tae dung nay tuang duang atorvastatin 10 mg/kg.
Dua tren eo so Iy luiin ciia Y hpe eo truyen, la Sen va Ha thii 6 do eo tiem nang digu tri cae benh ly Uen quan d&i roi loan ehiic nang tang Ty, Ty mat kien van sinh thap, uat ket thanh dam do nguyen nhan am thuc thuoc nhom bat noi ngocii nhan. Ket hpp 2 dupe Ueu nay co the giam sinh dam, giam tinh trang dam thap tac tro trong ca the, day la mpt trong nhiing nguyen nhan hinh thanh berth ly dam th% theo YHCT.De kiem ehiing gia djnh nay, can co nhiing nghien ciiu danh gia hieu qua ciia che pham phoi hpp la Sen - Ha thii 6 do tren benh nhan roi loan Upid mau the Ty h u dam thap, dam thap tac tro' kinh lgc do an uong nhieu chat beo ngpt
KETLUAN
Nghien ciiu nay da xac dinh dupe Ueu Dma<
cua cao LS - HTO la la 8,99 g cao/kg, tuong duong lieu 270,39 g dupe Ueu kh6/nguai 60kg, gap 34 Ian so voi lieu sii dung tren nguoi.
f***: p < 0,001); Khac biit co y nghia thong ki so vai Tren ehupt nhat trangtang Upid mau ngoai sinh do cholesterol (uong 25 mg/kg), ket hop eao la Sen voi cao Ha thii 6 do ty le 2-1 voi (cao la Sen 0,8 g/kg, cao Ha thii 6 do 0,4 g/kg)c6 tae dimg lam giam nong dp cholesterol toan phan, TG va LDL-C, tuang duong voi nhom uong atorvastatin 10 mg/kg. Khi sii dung Ueu thap hon, cao la Sen 0,4 g/kg va eao Ha thii 6 do 0,2 g/kg, thi thuoc thu nghiem dii the hien tac dpng giamTG
TAI LIEU THAM KHAO
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tac di,ing dieu hoa h p d mau ciia bgt say phun tir dai hoa Bijt gifun (Hibiscus sabdaiiffe L Malvaceae) ti@n chu^t nhat. T^p d l i Y Hpc TP HB Chi Mmh, t^p 17, so 1 tr 369-372.
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8 Wu C-H, Yang M-Y, Chan K-C, Chung P-J, O u T-T, Wang C-J (2010). Improvement m high-fat diet-mduced obesity and body fat accumulation by a Nelumbo nudfera leaf fiavonoid- n d i extract in mice J Agric Food Chem, 58(11): 7075-^81.
Ngdy nhan bai baa- Ngiy phdn biin nhan xet bai bt Ngdy bai bao dum: Sang:
30/08/2016 20/09/2016 25/11/2016
