Nghidn CCFU - Ky thujfct
Nghien ciru dich tac dung cua escitalopram trong dieu tri tram cam dtfa vao ky thuat tham tach micro va mo hinh dot bien gen
Nguyk Thinh Hdi', Gardier AlalnS Thdi Nguyin Hiing Thu' 'Tnr&ng Dgi HQC Dupe Hd NQI, Vi^t Nam
^Lah Dupe ly phdn tir ve than kinh. Truang Dgi hgc Paris 11. CH Phdp
D$t v^n 6h
Irhm cdm Id mpt trong nhOng b$nh xa hOi cd tdc d$ phdl tridn nhanh vi dnh hu-d^ng tn/c tidp d6n cudc sdng hang ngdy cua b$nh nhfln.
Nghifin c u u d u p e dlch Xic dyng ciJa thudc vd c o chd bdnh sinh cua trdm cam s§ gdp phdn khdng nhd trong vide cSi thidn hidu qud didu tri, hgn chd tac dyng khdng mong mudn cua thudc va gidm tinh trang khdng thudc tr§n b$nh nhan trdm cim. Escitalopram \i thudc chdng trdm cdm mai nhdt hidn nay dang du-g-c dp dyng trSn lam sang trong dieu tn bdnh trdm cdm d cdc nu-d-c Au-My"^' Cdc nghifin ci/u trdn in vitro ddu chi ra rdng escitalopram c6 tdc dyng iJc chd chpn IQC tren kenh van chuyen thu hdi serotonin, Idm tang ndng dp serotonin ddn synap sau cua td bao thdn kinh, tu' dd cdi thidn cdc tndu chii-ng Idm sang cua tram cam'^'. Tuy nhien tren cdc nghien CLKU in vivo va lam sdng, di'ch tdc dyng cua escitalopram van thyc s y chua cd nhieu bdng chung dupe khang dinh. Vi vgy myc dich CLia nghien c u u nay nhdm khdng djnh th6m gid tri khoa hpe ve dich tdc dgng cua escitalopram tren in vivo trong dieu tri trdm cam d y a trdn 2 ky thuat mdi Id : thdm tdch micro vd mo hinh chupt dot bien gen loai bd dfch tdc dyng nghi ngd ciia thudc chong tram cdm, cu the la kenh vgn chuyen thu hdi serotonin.
D o i t L P a n g v a p h i p c n g p h a p n g h i e n
D o n g vat t h i n g h i e m
Chupt d y e binh t h u d n g vd ehuot dot bien gen khdng cd kenh van chuyen thu hdi serotonin {knock out the serotonin transporter), khoang 4-6 tuan tudi, cd trpng lupng khodng 20-25 gam. Chupt dpt bien gen d u p e tgo ra bang
p h u a n g phdp gdy dpt bidn gen c6 chirc ndng tgo ra kdnh vdn chuydn thu hdi serotonin trong chudi ADN trdn chudt m^. Sau thd hd F1, cd 50% ehudt con mang gen ddt bidn khdng cd kdnh vdn chuydn thu hdi serotonin Dd phSn tdeh nhdm chudt ddt bidn gen, cdn su dyng p h u a n g phdp PCR (polymerase chain reaction) xdc dinh kidu gen cua tirng chudt trong thd hd F1'^'. Tdt cd chudt d u p e nudi vd kidm d|nh d u 6 l cdc tidu ehudn rdt nghidm ngdt (Moi nhdm chi cd 3-6 con, nhidt dd phdng khodng 22- 23*'C, phdng d u p e cung cdp dnh sdng 12/24h, cdc hd thdng nud'e udng vd thuc dn day du).
T h u d c SIP di^ng t r o n g n g h i e n ci>u Escitalopram oxalat (thude bdn quydn cua Cdng ty Lundbeck, Dan Mgch) d u p e nghidn cuu trdn chudt bdng cdch tidm vdo mdng byng vdi cdc lidu 8 mg/kg, lidu ddi chung dupe si> dyng bdng placebo (gid dupe). Lidu ndy dupe lya chpn d y a vdo nghidn c u u t h y c nghipm da dupe cdng bd trudc ddy'^.
KT t h u ^ t t h d m tach m i c r o
KT thudt thdm tdeh micro da dupe md ta trong cdc nghidn CLPU tnj-dc ddy'^'^'^. Cy the, chudt t h y c nghidm d u p e gdy me bdi cloral hydrat vdi lidu 400 mg/kg, tidm mdng byng. Sau dd, rgch ldp da d dinh ddu vd ru-a sach bdng nude oxi gid se nhin thay cdc vi tri xdc dinh tren mdng eipng (diem thdp, diem lambda). De xac dinh mpt vi tri cd djnh trong vung vd nao, ti>
diem thop thay ddi theo tpa dp (tien phia tru6'c:
+1,6 mm, sang bdn phai hogcsang ben trdi ±1,3 mm), sau do khoan 1 Id trdn mang a i n g cua v6 nao. Tai vi tri ndy se cay kim tham dd vao viing vd nao bang mdy bdn t y dpng vdi chidu sau co djnh Id: -1,6 mm (CMA7 model, Stockholm, Thuy Dien) cho phep thu hdi serotonin {5-HT) t?i
TAP CHI DUgfC HOC - 08/2012 rSO 436 NAM 52)
• Nghien C I F U - Ky thugt cdc khe synap te bdo thdn kinh trong viing v d nao. De cd djnh kim cdy edn do mOt Id-p keo gdn kim v d i mdng cung cua ddu chudt. Sau dd dua chupt v d o phdng hdu phSu, chdm sdc cho ddn khi tfnh d§y. Chupt se dd qua ddm eho mpi hogt ddng d u p e t r d l^i binh thudng. Sdng hdm sau, lai mpt ddu ddy kim se d u p e truydn mdt lo^i djch d u p e pha chd cd thdnh phdn vd ndng dd t u a n g d u a n g vai dich ed trong vd nao cua chudt (gdm. NaCl 147 mM; KCI 3,5 mM; CaClj 1.3mM;
MgCl2 1,2 m M ; NaH2P04 1,0 mM) v d i tdc flO truyen Id 1,0 /il/phiit qua mdy truydn tdc dd chdm (CMA100, Thyy Didn), trong khi dd chudt van di chuydn, an, udng v d hogt ddng binh thuo-ng. C u 20 phut se idy mdt lupng dich (thd tich 20 |il) de dem dinh lupng xac d|nh ndng dp 5-HT bdng mdy sac kl ldng hi^u ndng cao (HPLC), cpt C18, detector didn hda. Mdi chudt lay 4 mau lam diem ehudn, sau do chudt se d u p e tiem dud-i mang byng bdng placebo hodc ESC vP\ lidu 8 mg/kg vd tiep tuc lay ede mau trong vdng 2 gio' sau didu tri. Nghien cifu dupe t h y c hien lai tren chudt binh thuang va chupt dot bien gen khong cd kSnh vgn chuyen Ihu hdi serotonin trong mdi nhdm de lay gid tri trung binh, t u do thu dupe duo'ng cong dpng hpe ndng dp 5-HT (fmol/20fil) theo thai gian (phiit).
XLF ly ket qua
Phan tich cac gia trj thu dupe bdng phan mdm thdng ke y hoc StatView/ 5.0. (Abacus Concepts, Berkley, CA, L>SA). S y khdc nhau cd y nghTa thdng ke cua cac gia tri AUG (dipn tich d u d i ducmg cong ndng dp 5-HT theo thai gian) d u p e SU" dung bdng kidm dinh t-student, hoac
phdn tich s y bidn thidn mdt bidn (bidn didu tri) (One-w/ay ANOVA) vd-l kidm dinh Fischer, Gid tri khdc nhau ed y nghTa thdng kd ndu p < 0,05.
Ket qud nghien ciru
Ndng dd trung binh c c bdn cOa serotonin {5-HT) trong vd ndo trdn chudt ddt bidn gen vd chudt binh thud'ng
Ndng dd trung binh ea bdn cua 5-HT {fmol/20jjl) trong viing v d nao trdn chudt ddt bidn gen v d chudt binh thudng xdc dinh Idn lupt Id 14,2 ± 2,8 (n=20) v d 2,8 ± 0,6 (n=22). Cdc gid tr| ndy dupe tinh trung binh tif 4 didm chudn/ehudl v d dupe tidn hdnh trdn nhidu chupt thyc nghidm (n Id sd chudt thyc nghidm).
Hl^u qua cua escitalopram tren ndng d p serotonin (5-HT) t r o n g v 6 nao trdn c h u p t binh t h u v n g
Chupt d u p e tiem dudi mdng bung bdng gid dupe (paleebo) hodc escitalopram (ESC) v a i lieu 8 mg/kg Cac gia tri ndng dp serotonin (5- HT) dupe tinh trung binh cho moi nhdm (9-11 chudt). S y dap irng lieu cd tac dung ciia ESC tren ndng dp 5-HT trong viing v d nao chupt thuc nghidm duac mo ta trong hinh 1. Phan tich s y bien thien mpt bien (One-way ANOVA) tren ndng dp 5-HT trong vung v d nao thong qua cae gia tri di^n tich duPi duang cong ndng dp 5-HT theo thdi gian (AUC) dupe tinh trong khoang thd-i gian 120 phiit sau khi tiem dupi mang byng bdng placebo hoac ESC, cho thay co s y tang len CO y nghTa thdng ke v d ndng dd 5-HT trong viec a d u tri bdng ESC (F(1,21)=10,64; p <
0,001).
Q I
-eo -40 -20 Placebo Esc 8 m g / k g
T h o i g i a n ( p h u t )
Hinh 1 : Hieu qua cua ESC tren nong do 5-HT trong viing vd nao chudt binh thwdng.
A: Duang cong ddng hoc. B: Giatridi^n tich duwduvfng cong (AUC) ("*: p<0,001)
TAP C ' '^'X'XCJw.O_r -
• Nghien ciru - Ky thu j t
Hl^u qud cua escitalopram trfin n6ng d ^ serotonin (5-HT) trong v6 n i o tr«n c h u f t d f t bidn gen
T u o n g t v , chuOt d^t b i i n gen SuKfc tiSm d u d i mdng bgng b i n g paleebo ho$c ESC 8 mg/kg.
Cdc gid trj nJng dO 5-HT Suqio tinh trung binh cho m6l nhiim (9-11 chuOt). SM' ddp ling llAu c6 tdc dyng cCia ESC trdn n i n g dO 5-HT trong vCing v6 ndo chuOt thi/c nghi$m dLFVC mfl td trong
hinh 2. Phfln tich e y bidn thidn mOt bidn (One- way ANOVA) trdn n i n g d i 5-HT trong vting vJ ndo thftng qua cdc gid trj dl^n tich dui>l d u i n g cong nAng dO 5-HT theo thdrl gian (AUC) aiKfc tinh trong khodng thdi gian 120 phOt sau khi tidm d i f d l mdng bung b i n g placebo ho$c ESC, cho t h i y khftng ci5 s i / khdc bl^t c6 y nghla thing kd v 4 n i n g dO 5-HT trong vISc d i l u trj bSng ESC ( F ( 1 , 1 9 ) = 0 , 8 7 ; p > 0 , 0 5 ) .
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Thd-i g i a n (phut)
Hinh 2: H/^u qui cua ESC trin ndng dd 5-HT trong vOng vd nio chudt ddt bi4n gen. A: Di/Png cong ddng hoc. B: Gii trj di$n tich duPi du&ng cong (AUC) (NS: khic nhau khdng c6 y nghTa thdng k6)
thdy, escitalopram kh6ng Idm tdng t h ^ ndng dij 5-HT trdn nh6m ehudt ndy. Oidu ndy eho thiy escitalopram chl cd the tdc dyng uc chd trfen kdnh vdn chuydn thu hdi 5-HT md kh6ng cdn or chd ndo khdc Idm tdng 5-HT trong vung vd ndo Bdn l u | n
Cdc nghidn cu-u in vitro s u dyng trdn td bdo thdn kinh ngud-i da chung minh rdng escitalopram Id thude chdng trdm edm co tdc dyng uc che mpt each chpn Ipc nhdt a^ thu hdi serotonin (5-HT) tgi kenh vgn chuyen thu hdi S-HT*^^. T u 36 d i n ddn lam tang ndng dp 5-HT t?i synap sau cua td bdo than kinh. giup cdi thi^n cdc tridu chung Idm sdng eua bpnh tram cam. Nghien cuu in vivo cua chiing loi tren chupt binh thuPng cung chi ra rdng, escitalopram c6 khd nang lam tang nong dd 5-HT tgi cdc khe synap cua te bao than kinh (290%).
Tuy nhidn muc dd tdng ndy eo the xuat phdt do escitalopram uc che kenh vgn chuyen thu hdi 5- HT, dan den lam tang ndng dp 5-HT hodc cung e6 the l u mpt sd ea che ndo d6 Idm tang ndng dp 5- HT tgi cdc khe synap cua te bao than kinh.
Tren chudt ddt bidn gen khong co kenh vdn chuyen thu hdi 5-HT, ndng dp 5-HT trung binh ca ban tai khe synap Id 14,2 fmol/20pl, cao gap khoang 5 lan ndng dp 5-HT trung binh c c bdn tren chupt binh thuPng (2,8 fmol/20pl). Dieu nay hoan toan dupe giai thich do tren chupt dpt bien khong con kenh van chuyen thu hdi 5-HT. dan den 5-HT tai khe synap khong the dupe thu hdi va luon luon a trang thai tang. Khi thyc hi?n dieu tri bdng escitalopram tren chupt dot bien vai lieu 8 mg/kg (lieu co hieu qua tren chupt binh thuang), ket qua nghien cuu cua chung toi cho
K e t l u $ n
VPi vide s u dyng kdt tipp 2 ky thudt mdi: t h i n tdch micro vd m6 hinh ddt bidn gen trdn in vivo.
nghidn cCm ndy eung cdp thdm edc d u lidu khoa hpe cd gid tri vd dieh tdc dyng chpn Ipc cua escitalopram trong didu tri trdm edm. Ti> dd gdp phdn gidi thich trdn Idm sdng vd mue dd g i ^ thidu tdc dgng khSng mong mudn oia escitalopram trong didu tri bpnh nhdn tram cam'^.
S u m m a r y
For the fad that in vitro ^udies have s/wwn escitalopram having a selective serotonin reuf^ke inhibitor, which increases serotonin (5-HT) in the synapse of neuron serotoninergic, and so improi^ng the clinical symptoms of depression; but, in addition.
the in vivo studies have really not much evidence, this study aimed at examining the effects (^
escitalopram on extracellular levels of serotonin ([5- HTJett) in the frontal cortex c^ freely moving wildtype (WT) and mutant mice laddng the 5-HT transput (SERT-/-) by using intracerebral microdialysis and mutation mouse model. The findings proved tfiaf- (1) in l y r mice, a single systemic administration d T A P CHI nii-fKi-uf^f- flfl^^o^^ff^^^-,^ ^^^ 51)
• Nghien ctru - Ky thugt
esctfatopram (8 mg/kg, intra-petitoneal) produced a significant increase in cortical [SHTjaa f « 290%); (2) however, escitalopram failed to increase cortical (5- HTJaa in SERT-/- mice. In conclusion, escitalopram may seledively inhibit the 5-HT transporter only, but not any oUier mechanism of increasing the cortical [5-HTIexi-
T a i l i $ u t h a m k h a o 1. Murphy D.L., Lesch K.P, (2008). "Targeting the murine serotonin transporter: insights into human neurobiology", Nat. Rev. Neurosci.. Feb; 9(2);85-96.
2. Nordstrom G., Danchenko N, Despiegel N..
Marteau F. (2012), "Cost-effectiveness evaluation in Sweden of escitalopram compared with venlafaxine extended-release as first-line treatment in major depressive disorder". Value Health, 15{2):231-9.
3. Nguyen H T , Guiard B P , Bacq A., David D.J., David I., Quesseveur G., Gautron S., Sanchez C, Gardier A. M. (2012), "Bockade of the high-affinity norepinephrine transporter (NET) by the selectve serotonin reuptake inhibitor esotaiopram: an in vivo microdialysis study in
mice", Br. J. Pharmacol, doi: 10.1111^1476- 5381.2012.01850.x. [Epub ahead of print].
4. NguySn Thdnh Hdi, Guiard Bruno, Gardier Alain, Thdi Nguydn HCing Thu (2012). "Thdm tdch micro, kT thu$t djnh lupng thudc dang ty do tai mA dfch tdc dyng: Nguydn tdic vd Cmg dyng trong nghidn ci^u thudc", T^p chl Dupe h<?c. 52(429), pp. 6-8.
5. Nguyfin Thdnh Hdi, Guiard Bruno, Gardier Alain, Thdi Nguydn HOng Thu (2012), "Nghidn cuu co chd tdc dyng mdi cCia escitalopram trong didu tr| trdm cdm dya vdo kj thudt thdm tdch micro in vivo", Tpp ch/Dupe hpe, 52(431). pp. 37-41.
6. Owens fwI-J-, Knight D.L., Nemeroff CB. (2001),
"Second-generation SSRIs: human monoamine transporter binding profile of escitalopram and R- fluoxetine". Bbl. Psychiatry, 50(5). 345-350.
7. Sdnchez C. Bergqvist P.B., Brennum L T , Gupta S., Hogg S.. Larsen A., Wiborg 0.. (2003), "Escitalopram, the S-(+)-enantiofner of otalopram, is a selective serotonin reuptake inhibitor with potent effects in animal models predictive of antdepressanl and anxiolytic activities", Psychopharmacd. 167(4), 353-362
Nghien cihi bao che piroxieam nano bang phirffng phap ket tinh
Nguyen Thi Mai Anh, Nguyen Van Long, Nguyen Tru-cmg Son Truang Dai hoc Duac Hd Noi
D a t v a n d e
Trong ngdnh duac, vi tri cua cong nghp nano hipn dang d u p e khdng djnh bdng nhOng thanh c6ng trong nghien ci>u mot sd dang thudc t u nguyen lieu nano. Nguyen lieu nay ve ea ban d u p e chia thanh 4 loai bao gdm: nano tinh the, nano polymer, nano lipid ran va liposom'^', Trong do, nano tinh the la dang d u a c coi la c6 nhidu uu diem nhdt ve do tan, tdc dp tan va thuan Ipi cho bao chd nhieu dang thudc^'''.
Piroxieam la mot d u a c chdt thupe nhom oxicam (giam dau chdng viem khong steroid) Tuy CO nhidu uu diem ve tac dung nhung hidu qua dieu tn cdn han chd do rdt it tan trong nuPe.
VPi mong mudn lam tang sinh kha dyng eua piroxieam nano, chung toi tien hdnh de tai
"Nghien cmi bao che piroxieam nano bing phuvng phap ket tinh".
TAP CHI Dl/OC HOC - OS/2012 (SO 4:ifi NAM S21
N g u y e n l i e u , p h i r v n g p h a p n g h i e n CLPU
Nguyen lieu
Piroxieam (Px), tieu chuan USP 32 (Trung Qudc), polyethylen glycol 400 {PEG 400), methylen clorid (MC), aceton (Ace), ethanol (EtOH) tinh khiet phan tieh (Trung Qudc), alcol polyvinic (PVA) tieu chuan USP 32 (Singapore).
Mpt sd dung moi hoa chdt tinh khiet khdc dung cho bao chd va phan tich.
Thiet bj
May khuay t u IKA-WERKE (Due), may khuay tdc do cao UNIDRIVE X I 0 0 0 (Due), mdy ly tam Sigma 3-18 K Satorius, may sieu am Ultrasonic LC 60 H, kinh hien vi Satonus KRUSS (Buc), may do quang phd UV-VIS U1800 HITACHI (Nhat), kinh hien vi dien tir quet FESEM Hitachi S-4800 (Nhat), kinh hien vi dien
