N g h i e n
CLFU- Ky t h u a t
days of administeration. LD50 was determined as 135g (116.38 - 156.60) extract/kg (p = 0.05) by oral gavage In mouse. As for sub- chronic toxicity,- Eurycoma longifolia J. at the dose of 4 and 12 g/kg rabbits' body weight decreased the number of red blood cells, blood level of hemoglobin and hematocnt, increased serum concentrations of the total cholesterols.
Tai lieu tham khao
1. Bo Trung Oam. Phuong phap xac cJinh 60c tinh
c i p cua thuoc. Nha xuit ban Y hoc. (1996).
2. Duang Thi Ly Huang, Nguy§n Tran Giang Huang, Tran Dtrc Phan, Danh gia hoat tinh-androgen va tac dung len hanh vi tinh due cua re cay ba benh (Eurycoma longifolia J.) tren chupt thuc nghiem. Sap cao h6i nghi nghien ciru sinh lan thir XV- Trwang dai hoc Y, Ha N6I. (2009),
3. Vien Duac lieu, Cay thudc va dpng vat lam thuoc d Viet Nam, NXB Khoa hoc ky thuat Ha N6I, (2004), tr 116-118.
4. Ang HH et al, Eurycoma longifolia Jack enhances libido in sexually experienced male rats.
Exp. Anim. (1997), 46(4): 287- 290.
r
Nghien eii'u tae dung ehong oxy hoa eua
r
nu voi {Cleistocalyx operculatus (Roxb.) Merr. et Perry. Myrtaeeae)
Tran Minh Ngoc', Doan Cao Sen', Pham Tuan Anh^, Do Quyen^, Phirang Thien Thirong^
K/e'/7 Kiem nghiem Thudc Tnmg trang ' Trird'ng dgi hoc Dtrgc Hd Ngi. ' Vien Dugc lieu
Dat van de
Nhtrng nghien eu'u khoa hpe da chtrng minh ring eae dang dxy hoat ddng nhu hydroxyl (OH"), anion superoxid (O2" ), hydro perexid (H2O2) va dxy dan (02^ singlet oxygen) ... gay ra qua trinh dxy hea trong ea the, lam biin doi sinh ting hgp lipid, protein va aeid nucleic. Ttr dd, d i n d i n nhiiu benh nguy hiim nhu: ung thu, xa vua ddng mach, Packinson, lao hea, tieu dud'ng va nhiiu benh khac '^', Vi vay, viee tim ra eae chit ehdng dxy hoa de bao ve ca t h i trud'c nguy ea tren dang la xu huang nghien etru etia t h i gid'i'''.
Cay v i i (Cleistocalyx operculatus (Roxb.) Merr. et Perry) thudc hp Sim (Myrtaeeae), la eay mpc dai hoae duge tring, phan bo c- Thai Lan, Viet Nam, Lae va Trung Quic. Ttr lau, nhan dan da dting nu v i i lam nud'c uing h i n g ngay vd'i tac dung thanh nhiet, tieu thi/e '^'. Cac nghien CU'U trud'c day ehe thiy trong nu v i i ed chtra eac thanh phin hoa hpe nhu: chalcon, triterpen, terpen '^ '*' vai boat tinh sinh hpe nhu ehdng u (antitumor), u'c e h i enzyme ATPases '^ ^l Nhung hien chua ed tai lieu nae nghien etru tae
dung chong oxy hoa va sw lien quan tae dung vai ham lugng phenol toan p h i n etia nu v i i . Vi vay, nghien etru v i boat tinh nay gdp phin bo sung them tac dung sinh hpe va giai thich v i edng dung etia nu v i i dting treng dan gian.
Nguyen lieu va phu'O'ng phap nghien CLPU
Hoa chit, thuic thic, nguyen lieu
+ Nu v i i : Ouge mua 6' thj trud-ng (74 Hang Chiiu), edn ban str dung. M i u duge dung trong ttii nilon kin.
+ Nguyen lieu ddng vat: Nao ehupt nhit tring thuin chting (20 ± 2g) khoe manh duge nudi d"
phdng thi nghiem Truo'ng dai hpc Dugc Ha Ndi, Oai hpc Quic gia Chungnam, Han Qudc.
+ Hea chit: T i t ea eac hea c h i t dting treng cae thi nghiem d i u duge mua ttr cdng ty Aldrich& Sigma, Hoa Ky.
Phuang phap nghien eii'u Chuan bj mau thir
Liy 50g nu v i i dun each thuy vd'i methanol (150ml x 3lin). Gpp djch chiit, duoi h i t dung
TAP CHi DUOC HOC - 6/2011 (SO 422 NAM 51)
35N g h i e n CLFU - Ky t h u a t
moi d 50° C, dudi ap s u i t thap d i n khe duge eao methanol toan phan. Lam nhu tren vd'i vd'i dieh chiit nuae, duge eao nud'c toan phin, Cao methanol va cao nud'c toan p h i n duge pha loang trpng methanpl d i thtr hpat tinh,
Tac dung chong oxy hoa
Thw tac dung loai bo gdc DPPH thee phuang phap da dugc mc ta bd'i Phuang Thien Thuang va cdng su '''. Hut ehinh xae 5,0pl dung dieh m i u thu 6' eae ning dp khac nhau ehe vao 96 dTa (96 well plates), them ehinh xae 195,0 pi dung djch g i e tw do DPPH 150 pM, Oe treng t i i 30 phtit, do dp hap thu quang 6' bud'c sdng 520nm tren may Microplate Reader, Mdi m i u duac thtr d' 5 nong do khae nhau, Tae dung ehong dxy hea etia mau thtr duge xae djnh qua gia tri IC50 (ndng dp eua m i u thtr loai bo duge 50% gee ti/ do DPPH). Str dung a-toecpherol cateehin lam mau ddi chtrng.
Thd" tac dung dap tat gdc ABTS'*: Theo phuang phap danh gia tae dung dap tat gdc ABTS"* '^' hdn hgp phan trng gdm 490 pi dung djch g i e ABTS'* va 10 pi dung djch thtr. Sau dting 6 phtit, do dp hap thu quang 6' bud'c sdng 734 nm. Dung Trolox lam chuin g i e . Tae dung dap t i t g i e ABTS'* dugc bieu thj bing he s i tuang duang vdi tae dung ehdng oxy hoa etia Trolex a ning dp 1 (pg/ml) (TEAC; Trolox equivalent antioxidant capacity)
Thw tac dung ire chd qua trinh peroxy hoa lipid mang td bao nao chugt: theo phuang phap danh gia tac dung wc e h i qua trinh peroxy hoa lipid duge mieu ta bd'i Paul Cos va cpng sw '^'.
H i n hgp phan trng bao g i m : 10 pi dung djch m i u thtr, 340 pi dem kali phosphate 50 mM (pH 7,4), 100 pi dung dieh nao ehupt, 25 pi s i t sulphat 5mM va 25 pi ascorbic acid 10 mM; trdn d i u , tl d- 37°C treng bdng t i i 1 gid'. Them l i n lugt 250 pi trieloroaeetic aeid 20% va 250 thiobarbiturie aeid 0,67% trong natri hydroxyd 0,2mM, Oun each thuy sdi 30 phtit, lam lanh treng nud'c da, ly tam d" 5000 vdng/phtit trong 5 phut. O P dp h i p thu quang ctia dung djch thu duge o- bud'c sdng 532nm. Tac dung trc c h i qua trinh peroxy hea lipid eua m i u thtr duge danh gia thdng qua gia trj IC50 (ning dp u'c e h i 50% qua trinh peroxy hea lipid).
ThCr tac dung uc chd enzym lipoxygenase theo phuang phap Tea va edng si/'^'. Trong hon hgp phan ti-'ng ed: 5 pi dung djch thtr, 485pl dem
36
Tris - HCI 100 mM (pH 7,4), 5 pi linoleie acid 4 mM va 5pl lipoxygenase 10000 Unit. Trpn deu, dp dp hap thu quang d' buac sdng 234 nm sau dting 5 phtit. Tae dung trc c h i enzym lipoxygenase etia m i u thtr duge danh gia qua ning dp tre e h i 50% (IC50)
Xac dinh phenolic toan phan
Ham lugng eae c h i t phenolic dugc xac dinh theo phuang phap cua Gov/-Chin va cpng si/ ''"' dting galie aeid lam chuan. Chuan bj dung djch cao methanol nu v i i n i n g dp 0,5 (mg/ml) va dung djch cap nud'c 0,5 (mg/ml) trong methanol.
Htit ehinh xae 50 pi dung djch tren eho vao 900 pi natn eacbonat 2%, l i e d i u trong 2 phtit, them 50 pi Folin - Cioealteu de yen trong tdi 1 gid, Oo dd h i p thu quang 6' budre sdng 750 nm.
Ham lugng phenolic toan phin dugc tinh theo he s i tuang duang vd'i aeid gallic.
Ket qua nghien
CLPUva ban luan
Tac dung loai bo goc DPPH (DPPH Radical - Scavenging activity):
Kha nang loai bd gee DPPH etia djch chiet methanol (MeOH.ex) va dieh chiet nude (H20,ex) dugc danh gia thdng qua gia trj IC50, K i t qua a bang 1 cho t h i y tac dung loai bd gdc DPPH ctia djch chiit nud'c manh han etia djch chiit methanol vd'i gia trj IC50 etia djch chiit nud'c va djch chiet methanol l i n lugt la 22,5 v^
77,8 (pg/ml). Tuy ed t h i p ban c h i t ddi chtrng cateehin 7,0 (pg/ml) va a-toeophercl 14,5 (pg/ml), nhung tac dung leai bd g i e DPPH etia cac djch chiit nu vci la kha manh.
Tac dung dap t i t gdc ABTS'*:
Xac dinh gdc ti/ do cation ABTS'* [2,2'- azino-bis-(3-ethyl-benzethiazolin-6-sulfonie aeid)] la mdt trpng nhtrng phuang phap quang pho ea ban danh gia tong tae dung ehong oxy hoa etia dung dich mpt chit, dieh chiit hay ctia dd udng '^'. K i t qua 6' bang 1 eho t h i y ea 2 djch chiit d i u cd tac dung dap t i t g i e ABTS'* thdng qua vd'i he s i tuang duang vai tae dung ehong oxy hoa etia trolox (TEAC) 6' ndng dp 1 (pg/ml) l i n lugt la 0,98 (pg/ml) va 0,57 (pg/ml) eua djch chiit methanci va djch chiit nud'c so vd'i mlu d i i chtrng la a-teeopherol 0,31 (pg/ml) va cateehin 2,56 (pg/ml). Tae dung dap t i t g i e [w do cation ABTS'* cua djch chiit nu v i i t h i p ban cateehin va cao han a-tecopherol. Djch chiit nuae cd tae dung manh ban djch chiit methanol la vi ham lugng phenelic toan phin
TAP CHi DUOC H O C - 6 / 2 0 1 1 r s 6 47? N A M «1^
N g h i e n CLFU - Ky t h u a t
treng dich chiet nuac eae han dieh chiet methanol
Tac dung trc che qua trinh peroxy hoa lipid mang te bao chugt
Qua trinh peroxy hoa lipid tao ra cae gdc tu do, gay dde cho ea the. Do dd, trc che qua trinh peroxy hoa se lam giam nguy ea mac benh. Ca dieh chiit nud'c va methanol d i u cd tac dung wc ehe qua trinh peroxy hoa lipid manh, Ket qua duac the hien trong bang 1 vai ning dp wc che 50% qua trinh peroxy hoa ctia dieh chiit nuae va methanol l i n lupt 8,4 (pg/ml) va 7,1 (pg/ml) so vd'i chit del chtrng cateehin 5,6 (pg/ml) thap ban nhung eao ban a-toeopherol 16,3 (pg/ml) . Nhu' vay, cd the thiy tae dung tdt etia eac dich
Bang 1: Tac dung chdng oxy hoa cua dich chidt
chiet nu vdi treng phdng ngu'a va giam nguy ea m i e benh do qua trinh peroxy hoa lipid gay ra.
Tac dung u'c che enzyme lipoxygenase Enzym lipoxygenase thude hp enzym dioxygenase khdng chu'a s i t (non - heme iron - containing diexygenases), phan b i rdng rai trong ea dpng vat va thuc vat Chting tham gia qua trinh chuyen hoa acid arachidonie, gay ra oxy hoa mang lipid etia t i bao. Aeid arachidonie ddng vai trd quan trpng trong qua trinh viem. Qua trinh oxy hoa lipid gay xa vua dpng mach, chtr'ng huyet khoi va tham gia vao eae qua trinh benh ly nhu viem khdp va ung thu. Bd'i vay, chit wc che enzym lipoxygenase ed kha nang dieu trj eac benh nhu: viem, ung thu, mat kha nang mien djch'®'.
nwac va methanol cua nu vdi
Tac dung tren goc tw do Tac dung wc che, IC50 (pg/mi)
Mau thir DPPH
IC50 (pg/ml)
ABTS*
TEAC (pg/ml) Peroxy hoa lipid Lipoxygenase MeOH.ex
H20.ex Cateehin^
a-tocopherol''
77.8 ±0,85 22,5 ±0,62 6,0 ±0,48 14,5 ± 0 , 9
0,57 ±0,03 0,98 ±0,04 2,55 ±0,02 0,31 ±0,05
7,1 ±0,7 8,4± 1,1 5,6 + 1,5 16,3 + 0,9
75,3 ± 1,2 145,1 ± 2 , 5 7.6 ±1,1
b
m
Chat doi chirng: Am tinh: TEAC: He so twang duang vai tac dung chong oxy hoa ciia trolox a ndng d6 1 pg/ml: Tat ca thi nghiem dupc tien hanh 3 lan. S6 co nghla ± SD do lech chuan
O bang 1, eho thay tac dung ti'C e h i enzym lipoxygenase etia djch chiet methanol va nuac duac t h i hien bing gia trj IC50 l i n lugt 75,3 (pg/ml) va tren 145,1(pg/ml) so vd'i ddi chtrng la cateehin 27,5 (pg/ml). Djch chiit methanol ed tae dung manh ban dieh chiet nu'd'c, vi trong nu vdi cac dan ehat etia chalcone la thanh phan hoa hoe chinh '''•^'.
Ham lugng phenolic toan phan
Nhtrng nghien cu'u khoa hoc da chtr'ng minh ring polyphenolie trong djch chiet etia eay ed tae dung dap t i t eae gdc tw de va ching oxy hea '^°'. Ham lugng phenolic toan phin treng dieh chiit nuae va methanol eua nu vdi tuang duong vd'i acid gallic l i n lugt la 352,7 (mg/g) va 175,6 (mg/g). K i t qua duge bieu d i i n d- hinh 1 eho thiy ham lugng phenolic toan phin trong dieh chiit nuac eae han 2 l i n djch chiit methanol. K i t qua nay phti hgp vai cac phep thtr tae dung ching dxy hoa tren va eho thay m i i tu'O'ng quan giCra ham lugng phenolic toan p h i n vd'i tac diung ching dxy hoa cua djch chiit nu voi.
Hinh 1: Ham Iwgng phenolic toan phin (twang dwang vdi acid gallc)
K§t luan
Djch chiit methanol va nud'c etia nij vol da duge danh gia tac dijng ching oxy hea tren in vitro bing eac phuang phap thi nghiem nhu thtr tae dung loai bd g i e DPPH. g i e cation ABTS'*, ire c h i qua trinh peroxy hoa lipid mang t i bao va wc c h i enzym lipoxygenase. K i t qua thu duge ttr eae thi nghiem tren cho thiy hoat tinh chdng dxy hoa manh etia ea hai dieh chiit nu v i i , Djch chiit nuac ed tae dijng dap t i t g i e ti/
do DPPH va ABTS'* manh han djch chiit methanol vi ham lugng phenolic tean phin trpng dich chiit nuo-e cao han djch chiit
i"vii/~».'i^ iir\i~^ ^ / m i i / C O J22 iN.A.M 51) 37
Nghien
CLFU- Ky thuat
methanol. Tae dung u'c ehe qua trinh peroxy hea lipid va lippxygenase etia djch c h i i t methanol t i t b a n dich c h i i t nud'c, d i i u nay phu h g p vd'i nhtrng nghien etru v i thanh p h i n heat c h i t ctia nu v i i . K i t qua nghien etru nay g d p p h i n . l a m sang to kinh nghiem str dung nu voi treng dan gian.
Summary
The antioxidant activities of methanol and water extracts of the buds of Cleistocalyx operculatus Mert et Perry collected in Vietnam were investigated. These extracts showed possessing free radical scavenging activities against DPPH (with IC50 values of 77.8 and 22.5 pg/ml, respectively) and ABTS'* (TEAC 0.98 and 0.57 pg/ml, respectively). Fudhermore, both extracts remarkably inhibited either lipid- peroxidation of rat brain homogenate (ICso 7.1 and 8.4 pg/ml, respectively) or enzymic activity of lipoxygenase (ICso 75.3 and 145.1 pg/ml, respectively). These findings proved the ethnomediclnal use of C. operculatus as tea in Vietnam really beneficial to prevent oxidative stress-related diseases.
Tai lieu tham khao
1. Thuong, Phuong Thien; Na, MinKyun; Dang, Nguyen Hai; Hung, Tran Manh; Pham Thanh; Thanh, Tran Van; Nam, Nguyen Hai; Thuan, Nguyen Duy;
Sok, DaiEun; Bae, KiHwan, Antioxidant activities of Vietnamese medicinal plants. Natural Product Sciences (2006), 12(1), 29-37.
2. 0 6 Tat Lpi; Nhung cay thuoc va vi thuoc Viet Nam, NXB Yhgc, (2004), 423.
3. Le Thi Anh Dao, Nguyin Xuan Dung, Hcang Van Luu: Thanh phan hoa hpc ctia cac phan khac
nhau ctia cay vdi (Cleistocalyx operculatus Roxb Merrt et Perry) a Viet Nam, T^p chi Hoa hoc, (1997), 53 (3), 47-51.
4. Ye Chun - Lin; Lu Yan-Hua; WeiDong-Zhi Flavonoids from Cleistocalyx opeiculatur Phytochemistry (2004), 65 (4), 445-7
5. Ye Chun - Lin; Liu, Jian - W e n ; WeiDong-Zhi;
Lu, Yan-Hua; Qian, Feng. Invivo antitumor activity by 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone in a solid human carcinoma xenograft model. Cancer Chemotherapy and Pharmacclogy (2005),55 (5), 447- 452.
6. Woo, Anthony Y.H.; Waye, Mary M,Y,; Kwan, H S , ; Chan, Melanie C Y . ; Chau, C.F.; Cheng, Christopher H.K. Inhibition of ATPases by Cleistocalyx operculatus. A possible mechanism for the cardiotonic actions of the herb. Vascular Pharmacology (2002), 38 (3), 163 - 168,
7. Re, Roberta; Pellegrini, Nicoletta; Proteggente, Anna; Pannala, Ananth; Yang, Min; Rice - Evans, Catherine. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology & Medicine (1999), 26 (9/10), 1231 - 1237.
8. Cos, Paul; Rajan, Padinchare; Vedemikova, Irina; Calomme, Mario; Pieters, Luc; Vlietinck, Arnold J.; Augustyns, Keen; Haemers, Achiel; Vanden Berghe, Dirk. In vitro antioxidant profile of phenolic acid derivatives. Free radical Research (2002), 36 (6), 711-716.
9. Tea Joung Ha an Isao Kubo, Lipoxygenase inhibitory activity of Anacardic acids. J. Agric. Food.
Chem. (2005), 53, 4350-4354.
10. Gow-Chin; Hsieh, Chiu - Luan. Antioxidant activity of extracts from Du - zhong (Eucommia ulmoides) toward various lipid peroxidation models in vitro. Joumal of Agricultural and Food Chemistry (1998), 46(10), 3952-3957
^ r
Nghien eiru dieu che
va tieu chuan hoa acid azelaic
Trirong Phirong, Tran The Vinh, Nguyen Huu Lac Thuy, Le Ngoc Tii
Dai hgc Y- Diroc TP. Hd Chi MinhDat van de
Aeid azelaic la mpt acid dicarboxylic no mach t h i n g d u g e tim t h i y trcng Itia m i , Itia mach va
mach nha, va d u g e tao ra do mdt Ipai n i m la Malassezia fudur(haY Pityrosporum ovale). Nd cd tac dung trj benh da, khang khuan va kieh thich