?C CONG NGHE SINH HQC TOAN QUOC 2013
KHAO SAT MOT S 6 HOAT T(NH SINH HOC COA DICH CHI^T METHANOL CAY sAi HO NAM (PLUCHEA PTEROPODA HEMSL) THU THAP TO" VUC'N QUOC GIA XUAN THOY, NAM BjNH
Hoang Thj Ld, Dao Van Tan, Ddo Thj Sen*^
Trvdng D9I h<?c Sifph$m Ha Ndi
T 6 M T A T
CSy sai hfi nam {Pluchea pteropoda Hemsl) thuOc ho Cfic {Compositea hay Asteraceae) la loai cay phan bfi khd phfi bien t ^ Vircm Quoc gja Xuan Thiiy, Nam Dinh. Day la loai c6 t i ^ nang cho nhihig chit co ho^t ttnh sinh hgc. Trong bii bdo nay, chfing tSi phSn tich ket qua thu n g h i ^ Vak naog chong oxi h6a, kha nang irc che vi sinh vat, vi kha ndng iic dil sinh tnrang tiiuc v|t ciia djch chiSt meth^ol tit cdc bfi ph|n cfia loai cdy iiay Ho^t tinh chfing oxi hoa dugc tfait theo phucmg phdp quet gfic ti^ do DPPH, kit tpid cho thAy tdt ca cdc djch chiet thu nghiem diu co hoat tmh chfing oxi hda. Trong do, dich chilt cfia Id sdi hfi co boat tinh n^nh nhat (IC50 Id 2,46 mg/ml) vd dich chilt cua thdn co hoat tinh ylu nhat OCjo = 70,38 mg/ml). Ho^t tinh khdng khudn dugc thu theo phuang phap khukh tdn th^h, ket qud cho thay cdc d)ch cbilt co khd nang khdng duang tinh dfii vdi 2 lodi Basilus subtilis vd i^ibrio parahaemolyticus, manh nhat Id djcb chilt cua rS vdi Vibrio parahaemolyticus (D Id 11,3 mm). Cdc dich chilt co khd ndng khdng dm tinh doi v6i Escherichia coli. Vibrio ovan, Staphyloccus aureus. Cdc djch cbilt v6i methanol cfia cdc bfi phan cay sdi ho nam diu c6 khd nang lie chl sinh tnrdng it mdm h^tt cdi cii, tit 69-73% tfiy timg bO ph|n. .
7^ khoa: Sdi hh nam, DPPH, ure che vi sinh v|t, hoat tinb sinh hgc
MdTDAU
Vl^t Nam nim hong khu v^c nhi$t <36i gid mCia, da d^ng ve cdc h$ smh thdi trong dd bao g6tn cdc sinh thdi bt^n va ven bd vdl thanh ph4n thvc v$t phong phii. da dgng. Ti^m nang ay vln dang dugc khai thdc vd nghiSn cmj.vd thi,rc sg da trd* thanh mgc tieu nghien ctru ci^a nhieu nhd ktioa hQC. Trong nhOng th^p kl qua, thu6c chOa t)$nh co ngu6n goc t6ng hg'p tuy difgc sCr dyng nhi^u song ngu'&i ta ngay cdng nhgn ttiiy mat b'd! cOa n6 vd ngdy cdng tdp hung khai thdc vd nghi3n ci>u cdc lo$i thu6c co ngudn goc thidn nhiSn. Ket qua Id trong nhOng nam gin ddy rat nhiSu thuoc chOa benh c6 ngu6n g6c thdo du'grc da ra dcri vd th^c sg d6ng gdp khong nho vdo vi#c hi^n dgi twa nln y hgc c6 truykn etia Viet Nam.
Trong cac h$ sinh thdi ven bidn, h>ng ng^p mgn Giao Thiiy thuOc khu h$ viFon Qu6c gia (VQG) Xudn ThQy - khu Ramsar du'gc c6ng nhdn dku tidn t^i Vigt Nam id ngri hi>a hgn dly tiem ndng ve dugc ligu de nghidn cii'u vd khai thdc.
Thi^c vgt rCrng nggp m$n chi>a cdc hgp chSt sinh hgc co hisgt tinh khdng vims, khdng l^uln vd khang ndm trong tg nhien. Tlig'c v$t ng^p mgn Id mOt ngu6n phong phii cua steroid, triterpenes, saponin, flavonoid, alkaloid vd tannin, phenol. Nhilu nghidn ci>u da cht ra rang cdc chat chi4t xuat ti> nhieu lodi cdy rirng ngdp mgn cd nhieu tdc dgng du'gc trnh nhu* khdng khuan vd chong giun sdn, chdng oxy hda (Kabaru etal.. 2001).
Cdy sai h6 nam {Pluchea pteropoda Hemsl) mgc hoang dai d m$t s6 viing ven bien, rirng ng^p man (Phan Nguydn Hdng efa/., 1999). Sdi h6 nam c6 tdc dgng I6n trong y hgc co huyen. R i thu'd'ng di/gc dCing chO-a ngogi cdm, phat sdt ndng hoi rdt, nhijc diu, khdt nu'dc, tire ngi^c, khd chju, Uiay cho sdi hd bic (Bupleum sience DC) (Dodn Thanh Tirdng, 2001). Ld cd hu'ong tham Uiu'&ng dCing dk xong, cdn dung d l chOa dau mdi Ju'ng, trj cdc vlt lodt c6 gidi d hdu, bd.
Ngodi ra cdn cd tdc dgng lgi tilu, dieu kinh. Mac dii da cd nhirng nghidn ct>u v l thdnh phan hda hgc (Eltta Sdo et al.
2012; Nguyln Thj Chung et al., 2003) nhirng kha ndng chong oxy hda, kha nang irc c h l vi sinh vdt, irc chl sinh tnfcmg chu'a du'gc quan tdm nhilu. Chinh vi vgy, w^c xac djnh hoat tinh chong oxi hda, Cre chl vi sinh vdt, ire chl sinh trudng Id cin thilt d l xdc dinh cdng dgng du'gc tlnh vd danh gid diing du'vc gia trj cua lodi cdy do ti^ dd cd hu'dng phat triln vd khai Uidc.
Trong bdi bdo ndy, chung toi bin hdnh d i i l t nit djch chilt methanol tir cdc bg phgn khdc nhau ciia lodi cdy ndy nghien cii'u hogt tinh chdng oxy hda, khdng khuan vd irc c h l sinh tnj'dng
NGUYEN LI^U VA PHlfaNG PHAP N G H I £ N CIJU Nguyen lif u
Cdc bO phdn eiia cdy sdi ho nam {Pluchea pteropoda Hems!) dugc thu th|p tir rirng ng|p man Giao Thiiy, Nam Djnh Cdc Chung vi sinh v§t kilm tflnh: Escherichia coli, Badllus subtilis. Staph^ococcus aureus. Vibrio ovan, Vibno parahaemolyticus do Vifn V# sinh Dich t l Tnjng l/ong cung d p .
Thgc vdt de thu' khd nang d'c c h l sinh tru'dng: hat cdi cii {Raphanus sallvus L.).
Phuo-ng phdp nghien CLPU
Thu m i l l va chilt riit djch; Mlu tu-oi cdc bg phgn cua cay sdi hd nam {Pluchea pteropoda Hemsl) dugc thu thdp ^ rirng ngdp mdn Giao Thuy. Nam Djnh. Cdc mlu tuoi thu hdi v l dugc n>a sgrfi vd phdn loai cdc bg phdn. Mau dygc dem say Ithd d nhigt d0 SO^C.
Mlu da sly khd thi du'gc nghiin nhd, gdi trong giay Igc, ngam bong dung mdi methanol trong 24 gid, sau dd dem chiet bdng may soxhlet, djch chilt thu du-gc cho bay hoi hit dung moi thu du'gc d|Ch keo.
Thu hogt tinh chong oxi hoa dg* vao khd nang dap tat gdc tu> do (thu nghiem DPPH) (Vign Dugc ligu - Bd Y Te,
,r,nouv ~i i,hi nana tao ra cSc gic tv do >i*n trong dung djch ethanol b5o li6a. KM 2006); 1.1- dil)l«nvl-2-l)iciylliyd[a^ P P P " ) •» I ™ ^ E a n g am tmng hda hoSc t»o vay cio gbo tv do s§ ISm g a ,
trrroirrgrrc:..-^^^^^
« ^ 1995) dira vio dOdSi cua av(nigk(nhv6ngvoktiuSnD (mm).
s l a l . I9»3)airav»j V . . , H . „ .h,rona Dhiip cOa T r i n T i l n Oft va cs (1999): Hat cji ci Thft khS nang Oc ch« .Inh t " ^ ' • « ' * ^ ' f ' S S i ? t dS 6n dinh (30'C ) 1 t,o cSy mSm! NhOng c4.
(Rap/,a™s sa//™s L.) sau kh, '^^'^l^^^^^^'^ZZ^Sri cSy mSm diwo nhung tong 1 gift vie dich chii mSm c6 r i ding diu nhau d u j c * ' " " i ' * " J f , J ^ 3 * ^ ^ nhic lai 3 lan. Cay mim d l l chOng thay did.
S S S ' ^ i ^ l ' S ^ r J m Z ^ " r 2 ? g t " ' l i h i n . ^ ^ ^ ^ ^ ^ l % = (U-l-.)x10(WU
I %: phin hdm dc chl
L,; Dg ddi tnjng binh (cm) cila cdc r l d mau ddi chdng U: D$ ddi tnjng blnh (cm) aia cdc r l d miu thi nghidm S l ll$u dugc phdn tich trdn phin mim Excel vd SPSS.
K^T QUA V A T H A O LUAN Hogt tfnh ching oxy hda
n l tAch rhllt cdc chit trong thuc vdt ngu&i ta sir dgng cdc dung mdi khdc nhau nhu nude, methanol ethanol,
?^Sne ^ c dung n ^ S . STkh^nhau v l dO phdn cgc do dd sd dnh ^ l ^ ^ n g ^ ^ " ^ ^ « ^ ! " ; ' ' ^ S / L K ' I S S ^ c h g p S l t c6 hogt tlnh sinh hgc. Trong dd. methanol Id m^t dung mli thudng dugc su dgng d l tdch thu d ch S r i t ttid ffkhdo S hS^ ttnh sinh hgTbdi khd ndng hda tan tit, ddc bi$t Id cdc cdy rung nggp m$n thudng chua nhilu S r S ftT-S phUhemfcallSartins et al.. 2001). VI vdy, d l khdo sdt hogt tfnh smh hgc ban diu cua c^y sd, hfi n S dSng WdS IgaX-Tmelhanol Idm dung mdi d l tdch chilt Mlu khd cdc bg ph|n cua cdy dugc ngdm trong n^ttiano) 24 gld, sau dd den, chilt bing mdy soxhlet din djch chilt trong r6i bay her, thu cdn ch,et Cdn chilt dugc hda tan liing nude cit su dgng cho cdc th( nghi§m tilp theo.
D l thd hoat tlnh ching oxi hda. chting tdi tiln hdnh thd nghi#m theo phuong phdp quet goc tg do DPPH vd tlnh lugng djch cin thUt d l dc chl 50% glc tg do DPPH. DPPH Id glc h^ do cd mdu tlm nhd vdo dign td N chi/a ghdp ddi nhung sau khi phdn ung vdi oxi nguydn td cOa chit ddP tit glc \}f do se Idm gidm mdu tim dan din Idm gidm dd hdp thu * budc sdng 517nm. Trong nghidn cdu ndy, tnidc hit chOng tdi khdo sdt hogt tfnh chdng oxy hda d cdc ndng d | pha lodng djch chilt khdc nhau: 4, 10. 20, 50, 100 lln. Sau dd xdy dgng hdm hll quy de tlnh gid trj ICso. Kit qud thu dugc h-lnh bdy bong bdng sau:
Bdng 1. Ho^t tfr)h ctiAng oxy h6a cda djch chiit methanol cic b$ ph$n cda cdy BO phfin ICw (mg/ml) % acid ascorbic
Rfi Ttidn
0.64 3,38 7,71 0.47
Tu bdng cho thiy hogt tfnh dilng oxi hda ciia djch chilt td Id sdi t i l nam Id cao nhlt (ICso =2,46 mg/ml tuong duong vdi khd ndng qudt glc tg do cua 7,71% ascorbic acid). Tilp ^eo Id khd ndng ching oxy hda ciJa cdc djch chilt td ttidn sdi h i ( ICso =7,33 mg/mt). Khd ndng ch&ng oxy hda ttilp nhlt Id djch chill r l (ICK» = 50,94mg/ml). Nhin chung, khi ndng ching oxy hda ciia cdc djch chilt h> cdy sdi h i nam Id cd tllm ndng. So sdnh hogt tinh ching oxy hda cCia nhilu cdy hong chi Pluchea nhdn thiy id cua chi ndy cd hogt tfnh oxy hda mgnh. Kit qud nghien cdu trin cdy sdi h i nam phl^
hgp vdi cdc bdo cdo tr\/dc dd v l cdc lodi trong chi vd cd tiim ndng dng dgng r i t Idn. (Elita Scio et al., 2012; Kumar sf af.. 2011)
100 1
^ 8 0 ! _ ^ — — — ' S ^ V • 13.93ln(iil. 8G.54 o 60 ^ . - - ^ R>. 0.984 X *0 I
0 0-2 0.4 0 6 0.8 1 N6ngd4(9'ml)
100 -,
90 ] » 80 J * _ _ -
70 •) ^,^-'^ V=13.71ln(K)* 10.41
%^° \ / R' = 0.973 1. 50 J>
u 40 T ' a 3 0 1
« 20 H J 10 \
0 100 zoo 300 400 500 NSngae(rTig/miy""' Hlnh 1. Ttro^g quan glira ning d$ djch chl6t va-i tl 1$ % quit glc tg do
Hlnh 1 t h i hi§n mi^ hfcmg quan giua fi 10 (%) dc chl g l c DPPH vd ning do djch chilt Kit qua phdn tich cho thiy mli tucmg quan hjdn theo quy ludt ham logarithmic vdi R^djch chilt r l sal ho nam id 0,984, R^ djdi chilt Id sdi h i nam Id 0,973. Dilu ndy Chdng td fi t# dc chl gdc tg do DPPH vd ning d$ djch chilt cd moi tuong quan chgt che.
Qud blnh ho hip hilu khi c l ^ t l bdo thudng xuydn tgo ra cdc goc tu do td do hinh thdnh cac goc oxy phdn dng (reactive oxygen species, ROS) nhu hydroxyl (OH"), SuperoxHle(Oz ^'.) nitric oxide (NO") vd hydrogen penaxide (H2O2)- Cdc glc h/ do khi du fiii>a se gdy nhilu t i n hgi din t l t)do nhu Idm gidm tinh linh dgng cua mdng. cdc thy t h i enzyme mdng khdng hogt dgng cijng nhu cdc protein mdng... d i n din Idm d i l t t l bao. Cdc glc tu do con lidn quan din cdc n&l logn khdc nhu lao tida, ung thu, t)^nh fim mgch, tilu dudng, vidm khdp dgng fiilp, ddng kinh... Con dudng hi$u qud nhlt d l logi bd vd Idm giam tdc d ^ g cOa cac goc tg do gdy ra su du thda oxy nguyen h> Id ca chl bdo vi ching oxy hda. Chit ching oxy hda la nhung chat cd phdn dng b l gay chuoi tg do. Chat chong oxy hda cd t h i tri hodn, dc che ho|c ngdn chgn qud blnh oxy hda cda vdt lidu d l oxy hda tiang v i ^ quet cdc glc tu do va lam giam bdt oxy du thua.
Kit qud nghien cdu ben cho ttily, djch chilt methanol cOa cdy ndy r i t cd film nang de sd dung nhu mdt chit ching oxy hda. d thgc vgt, hoat ddng chong oxy hda thudng lidn quan tdi cdc hgp chat phenolic do chting cd chda hydro. Cdc nghiSn cdu v l thdnh phan cac cdy thudc chi Pluchea cOng cho ttily hdm lugng cdc chdt phenol chilm W 1$ cao (Kumar e/a/.,2011). Vi vdy, hoat ttnh chong oxy hda cua cdy sdi h i nam cd t h i cung thudc v l nhdm cac hgp chat phenol.
Hogt tfnh khdng khuan
Chilt xuit mettianol cua cdc bd phdn hoa, Id, thdn, r l cua cdy dugc kilm tra tinh khdng khuin trdn cdc chCing vi khuin E.coli. Bascillus subtilis, Vibno parahaemolyticus. Vibrio ovan. Staphylococcus aureus. S l lieu ttid hogt tlnh khdng khuin thi hl#n bong Bang 2.
Bdng 2. Ho^t tinh khdng khuan trin chung vi khu^n khdc nhau (vdng dc chl. O mm)
Bd ph^n
RS Thdn Ld Hoa
E.cdi B. subdiis 4,0 2,9 2,1
ChiJng vi k h u i n V. pamhaemdyticus
11,3 9,8 8.6 8,7
V.ovan S. aureus
Ghi cha: (-) thi hi$n khdng xdc djnh dugc vdng in chi
Td bdng kit qud cho thiy djch chilt td cdc b$ phdn khdc nhau cda cSy deu cd khd nang khdng khuin vdi khd ndng khdng Id khdc nhau. Cac djch chilt diu c6 khd ndng khdng du'o'ng tinh vdi 2 chung Bascillus subtilis vd Vibno parahaemolyticus, eae djch chilt c6 kit qud dm tfnh dli vdi ba chflng E. coli. V. ovan vi S. aureus. Djch chilt mettianol cua thdn vd r l sdi ho nam cd khd nang khdng mgnh hon djch chilt td Id vd hoa cua lodi nay. Dilu dd cho thiy cd cdc chit khdng khuin phdn bo khong ding diu trong cdy sdi h i nam. Trong ddn gian, khi sd dgng cdy sdi h i chda b$nh ngudi ta thudng su dung r l vd Id, vd dilu dd dudng nhu cd lidn quan tdi cdc chat cd tdc dung dugc If mgnh ho'n trong cdc bd phdn ndy.
Thulc khdng sinh dugc sd dgng d l ching Igi cdc vi sInh vdt gdy bdnh nhung di kdm vdi dd Id theo thdi gian vd cudng dO su dgng se xuit hign cdc loai d l khdng nd. Vi khuin ngdy cdng cd kha nang de khang vdi cdc logi khdng sinh thdng thudng vdi tic dg ddng bao dgng. Do dd. vigc flm kilm cdc chit cd hoat tfnh khang khuan mdi la rat quan trgng. Trong dd, cdc sdn phim tu nhien td Ihg'c vgt cd thi Id Iga chgn ttiay t h i thu vj. Kit qud budc diu v l hogt tfnh khdng khuin cfla djch chilt tho methanol cdy sdi h i nam da cho ttily tinh khdng vdi 2 chung Bascillus subtilis vd Vibrio parahaemolyticus. Sir dgng djch chilt ttid ndn sg cd m$t cfla cdc hgp chit cd tfnh khdng khuan c6 thi khdng nhilu. Vi v$y, d l cd ddnh gid chi filt, day dij han v l hogt tfnh khdng khuin cung nhu su dgng lodi cdy ndy bong vide khai ttidc sd dgng nhu thuoc khdng sinh c i n tach chilt cdc hgp chit tinh khilt han vd khao sat them cac chdng vi khuin gdy bgnh khdc
^ , ,_i B (.•«(!,.„.•.., M,.h ^uiAt ,^^tu I Hlnh 3. Kha nang dc che sinh trifdng re cdi cu cua djch chlSt Hlnh 2. Ho,! tlnh khing B. sMI^cu^ djch Mit mM^nol ^^, ^^^ ^^, ^^ „ . j , ^£ ti„ha„ sal h i ram. (B) cdc b$ ph^n. (+) khdng smh Chloramfenico! 0,4%, (-) nuwc cat , rhLmn
khi> InJng; (TS) djch chilt cua than sdi ho nam, (RS) D|ch chill °°' '^ ^ ^ da r i sdi h i nam; (LS) Dich chiet cOa Id sdi h6 nam, (HS) Djch chilt hoa sdi hi nam
HOI NGHj KHOA HUU u u n o ••
Kha aitia i>c cti^ sinh tnFimg thuc v$t
. _KA • K»,.A™,tt,„<-»at cua c4c didi * * t t i ( r nghldm, Chung 161 Bin h&ih mi, ngNSmli^
sau K.«. I, -* V. d.. * ^ , ; r j , r d ^ s^-^.ti ^iiiri'^sf^h?^g*sa?:^7*^^^^ j. *"Si
11,1 nghl#m. So sanh gli tn trang blnh * ' * " ??'™ | f ^ ' 2 " 1 ^ „ h dtlfcl dSi trang blnh cCia chliu dai rS mim cOa mk
5sst^.itr.Sn3M|Si^*?*^^^^^^
rir<^s-sth=;s^^g'<S'S.xs:"ftnT;?Ka^*---*''^*B^'-3
B i n , 3. H.^unh i,c ch* .Inh i ™ . g .1 ci,cai o i (I %) cua d|ch c h i a f t ^ n g l ^ B^phin R^
^Snt»^^**-'i'--prnS^^^^^^^^
S ^ d d ^ g r i l p h d n bdo ^ ? T S khd nang^flc c h l phdn bdo t i t cfla djch chilt cdy sd, h i nam cd ttil cd n U nang bong vide nghiSn cdu khai thdc tlm cdc hogt chat chong ung ttiu.
K ^ LUAN
Dich chilt methanol td cdc bd phgn cfla cdv sdi h i nam (Pfuchea ptenapoda HemslJ vua cd hogt tinh khdng khuin, vira I cd khd ndng dc chl sinh trudng ttigc vdt ding^thd, cd khd ndng ching oxy h ^ . Trong fld h j ^ t J l J ^ ^ a wty hda M Id Id mgnh nhlt (IC» Id 2,46 mgftnl), djch chilt cfla thdn ylu nhat (IC50 = 70-38 mg^ml Cdc djrt, ch.lt c6 khd r ^ khdna duong tinh d l l vdi 2 todi Basikis subtilis vd Vibrio parahaemolybcus. manh nhdt Id djch chidt cua ifi vd, Vitao parahaemolyticus (D Id 11,3 mm). Cdc djch chiet cd khd ndng khdng dm inh d l i vdi Eschenchia cdl, Vibno oi/ai Step/iytoccus aureus. Hogt tlnh dc chl sinh budng r l cdi cu {Raphanus salivus L.) cfla cdc djch chidt thu dugc Id dii mgnh (69.34 % din 72,87%).
K i t qud nghidn cdu ndy ggi # rang ddy Id kidi cdy t l m ndng d l nghifin cdu cdc hgp chit c6 hoat tinh anh hgc.
TAI U$U THAM KHAO
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BIOACTIVITIES OF METHANOL EXTRACTS DERIVED FROM DIFFIRENT PARTS OF PLUCHEA PTEROPODA HEMSL IN XUANTHUY NATIONAL PAEK.
NAM DINH PROVINCE
Hoang Thi La, Dao Van Tan, Dao Thi Sen Hanoi National University of Education
Pluchea pteropoda Hemsl of fiie &iiiily Asteracea, is a common species m Xuan Thuy National Conservation Paric, Nam Dinh Province with high potential for bioactive substances. In this study, mettianolic extracts fiom various parts, including root, stem, leaf and flower, of Pluchea pteropoda Hemsl collected fix)m Xuan Thuy National Park, Nam Dinh Province were evaluated for their antibacterial and antioxidative activites and inbitntion of plant growth. DPPH fiee radical scavenging activity on crude methanol extracts were evaluated. The maximum radical scavenging activity was observed for the extract from leaves (ICso value, 2.46 mg/ml) and minimum radical scavanging activi^ was shown by the extract fiom stems (IC50 value, 70.38 mg/mt). The antibacterial activity tested using agar difEusion method, the results show that the extracts are resistant to both Bacillus subtilis and Vibrio parahaemolylicus, the strong etiect of growth inliilntion on V. parahaenu>fyticus is root extracts (D = 11.3 mm). The extracts are not resistant to Escherichia coli. Vibrio oval, Staphyloccus aureus. All parts of the plants showed inhibition in ifae growth of seedling Raphanus salivus L. roots, rangjng from 65-72%, depending on parts of the plants.
Keywords: Pluchea pteropoda Hemsl, DPPH, antimicrobial activity, inhibition, bioactivitics.
Auttior for co^espondence: Dao Thi Sen, Email: [email protected]
