HP CHT KHNG KHUN, KHNG NM T THC VTHNG DN KHOA HC: Gs.Ts. CAO NGC DIP

NI DUNGGII THIU CHUNG V KHNG SINHNHM CHC C HOT TNH KHNG KHUNNGHIN CU THC VT KHNG KHUN TRN TH GII

I. GII THIU CHUNG V KHNG SINH

I. GII THIU CHUNG V KHNG SINHKhi nim chungPhn loiLch s khng sinhGiai on 1928 1945Giai on 1945 v sau

Khi nim chung nh ngha

Sinh hc Bn tng hp Tng hp

Vi khun Vi nm T boNgun gcc ch chuyn ha

Khi nim chung

Phn loiNhm lactam (penicilin v cephalosporin).Nhm aminosid hay aminoglycosid.Nhm cloramphenicol.Nhm tetracyclin.Nhm macrolid v lincosamid.Nhm quinolon.Nhm 5- nitro- imidazol.Nhm sulfonamid.

Phn loiDa vo cu trc ha hc

Thuc c ch tng hp vch t bo vi khunThuc c ch tng hp protein ca vi khunThuc c ch tng hp acid nucleicThuc c ch chuyn hoThuc lm thay i tnh thm ca mng t bo

*Phn loiDa trn c ch tc dngPhn loi

1. Khng sinh c hiu2. Khng sinh ph rng3. Khng sinh ph hp

Phn loi theo ph khng khunPhn loi

Lch s khng sinh

Giai on 1928 1945PenicillineSulfonaidGiai on 1945 v sauTm khng sinh miPht trin khng sinh c

*Khng sinh c pht hin giai on 1940-2000 *

Giai on 1928 - 1945Nm 1922: Alexander Fleming pht hin ra Lysozime c trong nc mt, nc mi, nc bt, dch v, ca ngi. Lysozime l 1 enzyme c tc dng c ch s sinh trng ca 1 s vi khun

*- Nm 1928: Alexander Fleming pht hin penicilline (t nm Penicillin notatum)- Nm 1929: A.Fleming cng b kt qu nhng lc u cha to c s ch ca d lun.

Giai on 1928 - 1945

Nm 1932: Sulfonaid c Gerhard Domard (ngi c) tm ra.SulfonaidGerhard Domagk (1895 1964)Giai on 1928 - 1945

Giai on 1928 - 1945

Nm 1934: Streptomycin c Selman Waksman v Albert Schatz tm ra

Nm 1939: H.Florey v E.Chain bng phng php ng kh chit tch ra c Penicilline.

PenicillineHoward Florey v Ernst Chain Giai on 1928 - 1945

Nm 1945: A. Fleming c gii thng Nobel v y hc v ng chia s gii thng ny cng H.Florey v E.Chain Giai on 1928 - 1945

Penicilline c a vo th nghim lm sng v ng dng cu sng cc thng binh trong Th Chin II.

Giai on 1928 - 1945

Giai on 1945 tr v sau1962, cc khng sinh tng hp , thuc chng vi sinh vt c bn cht quinolone c to ra.Tip tc ci tin trong tng lp khng sinh v thu c khng sinh c hot tnh cao hn nh -lactam.

S tin ha ca penicillin G

Th k 20, vic pht hin, pht trin v s dng cc khng sinh trong iu tr lm gim ng k t l cht do nhim khun.Nhng nm 1980 s khng sinh mi c a vo iu tr gim hn do vn chi ph cho vic pht trin v th nghim thuc mi.Tnh hnh khng khng sinh tng cao.

Hin nay, vic khng thuc ca vi khun phi c khc phc bng vic pht hin ra cc thuc mi. Tuy nhin vi sinh vt cng ngy cng nhanh khng thuc v tc li nhanh hn tc to c thuc mi ca con ngi. Do vy, cc nghin cu cn phi tp trung vo lm th no vt qua tnh khng thuc cng nh pht hin cc khng sinh mi c cc c ch hot ng khc nhau.

II. Nhm chc khng khun1. Phenol n v acid phenolicTo ra hng v v mu sc cho cc loi cy trng. Bo v cy chng li s tn cng ca cc loi vi sinh vt, cn trng v ng vt n c.

II. Nhm chc khng khun1. Phenol n v acid phenolic

Ngi gimC x hng

II. Nhm chc khng khunEugenolEugenol (tinh du inh hng) c xem l cht kim hm ng thi chng li c nm si v vi khun.

II. Nhm chc khng khun1. Phenol n v acid phenolicCatechol v pyrogallol c cho l c c tnh i vi vi sinh vt.

PyrogallolCatechol

II. Nhm chc khng khun2. Cc hp cht quinonesQuinones l vng thm vi hai nhm ketone.Quinones gy s bt hot v lm mt chc nng ca protein do hnh thnh phc khng phc hi vi cc axit amin ca vi khun.

II. Nhm chc khng khun2. Cc hp cht quinones* Cy c ch: Tc dng khng khun:Candida mycoderma, Corynebacterium diphteriae, Bacillus antracoides, B. anthracis, B. Subtilis, Hansenula anomala, Botrys cinereav nhiu loi nm mc, cha Herpes, tr ho, vim ph qun.

II. Nhm chc khng khun2. Cc hp cht quinones* Cy bch hoa x- Staphylococcus aureus, Streptococcus pyogenes, Pneumococcus, Mycobacterium tuberculosisEscherichia coli,- Cha cc bnh ngoi da, nhng vt lot, vt thng, cha gh.

II. Nhm chc khng khun2. Cc hp cht quinones* Mt s loi thuc h Gng v- Mycobacterium tuberculosis- Droseron cn c tc dng chng co tht, cha ho.

II. Nhm chc khng khun2. Cc hp cht quinones* Cy l mng

Nhm vi khun: Staphylococcus, Streptococcus, Brucella v Salmonella,Cha l lot, hc lo, cha ho, vim kh qun.

II. Nhm chc khng khun3. Flavonoid Cc flavonoid c hot tnh khng khun do chng c kh nng to phc vi cc protein ngoi bo v thnh t bo vi khun.Flavonoid cng a bo cng c kh nng ph v mng t bo vi sinh vt.

II. Nhm chc khng khun3. Flavonoid * Tr xanh

Catechine v hot c t gy bnh t ca Vibrio cholera 01, c ch enzym glucosyltransferase ca Streptococcus mutans, c ch hot ng ca Shigella v mt s vi khun, vi nm khc.

II. Nhm chc khng khun3. Flavonoid Mt s flavonid nh quercetin, hesperetin v catechin c th chng li virus type 1 (HSV-1) gy bnh mn gip, polyvirus type 1, virus type 3 gy bnh kh th tr.

II. Nhm chc khng khun3. Flavonoid

Galangin (3,5,7-trihydroxylflavone) c chit xut t cy tho mc sng lu nm Helichrysum aureonitens c hot tnh chng li vi khun Gram (+) cng nh nm si v virus, c bit l HSV-1 v coxsackie B type 1.

II. Nhm chc khng khun4. Tannin L nhng hp cht polyphenol c trong thc vt, c v cht c pht hin dng tnh vi th nghim thuc da v c nh lng da trn mc hp ph ca bt da sng chun.

II. Nhm chc khng khun4. Tannin Khi lin kt vi protein chng c th lm mt hot tnh ca cc protein chc nng. Cc protein ny c th l enzym, cc protein vn chuyn hay thnh t bo polypeptideTannin c th gy c cho nm si, nm men v vi khun.

II. Nhm chc khng khun5. AlkaloidNhng hp cht hu c c cha nit, a s c nhn d vng.C phn ng kim.Thng gp trong thc vt v i khi trong ng vt.C dc lc tnh mnh v cho nhng phn ng ha hc vi mt s thuc th chung ca alkaloid.

II. Nhm chc khng khun5. AlkaloidSolamargine: c trong cy mng h c.Chng li s ly nhim khi mc phi HIV, khng khun tt nht i vi 2 nhm Giardia v Entamoeba v khng cc vi khun tiu chy.

II. Nhm chc khng khun5. AlkaloidCy hong lin:

Berberin c tc dng vi vi trng gy bnh st rt, khng vi khun Shigella, t cu khun, nhiu vi khun Gram (-), Gram (+) v cc vi khun acid.

II. Nhm chc khng khun6. Terpenoid v tinh duTerpen: cht chuyn ha th cp da trn n v isopren. Khi cc hp cht c cha cc thnh phn b sung, thng oxi, n c gi l terpenoid.Tinh du thng l hn hp cc chc khc nhau: hydrocarbon, phenol, ru, aldehyde, acid, ester,nhng thnh phn ch yu l terpen v cc dn xut cha oxy ca terpen.

II. Nhm chc khng khun6. Terpenoid v tinh duCy thanh hao hoa vng:

Artemisin l mt secquiterpen chit xut t cy thanh hao hoa vng (Artemisia annua) c hot tnh khng st rt mnh, hot cht ny c nghin cu v phn lp t nhng nm 70.

II. Nhm chc khng khun6. Terpenoid v tinh duTinh du t cc loi s -Andropogon spp. c tc dng khng khun ch yu ln vi khun Gram m. Tc dng ca tinh du s t l thun vi hm lng citral.

II. Nhm chc khng khun6. Terpenoid v tinh duTinh du t cybch n-Eucalyptus globulusc tc dng lnBacillus subtilisv Staphylococcus aureusnhng km tc dng i viE. coli.Cineol trong tinh du l thnh phn c tc dng.

II. Nhm chc khng khun6. Terpenoid v tinh duTinh du ca mt s cy h Hoa miTinh du cy hng di (= hng qu) -Ocimum basilicumL. C tc dng khngSalmonella typhi.Tinh du cy hng nhu trng -Ocimum gratissimumL. C tc dng khng vi khun lao.

II. Nhm chc khng khun6. Terpenoid v tinh duMenthol c trong tinh du bc h c tc dng chn lc trn mt s chngBrucellanhB. melitensis, B. paramelitensis, B. suis.

II. Nhm chc khng khun7. Lectin v polypeptideThionin

Thionin l peptides thng c tm thy trong la mch v la m v bao gm 47 amino acid. N gy c n nm v vi khun Gram m v Gram dng.

III. THC VT KHNG KHUN CC VNG TRN TH GIISaudiMalaysiaBraziln Nam Phi

1. Saudi 1.1 S lcKho st kh nng khng khun, khng nm ca dch chit methanol t 9 cy thuc c ngun gc y hc dn gian Saudi.

1.2. Phng php tch chitX l, lm kh nguyn liu v xay nhuyn thnh bt.

Ngm nguyn liu trong methanol khong 72 gi, va ngm va khuy.Lc v lm bay hi dch lc thu dng cao thBo qun iu kin thch hp

1.3. Phng php th hot tnhHa tan cao vo methanol t nng 100mg/mlLc vi mng lc c kch thc lc 0.22mChun b vi sinh vt vo mi trng nui cy vi sinh vtBm vi sinh vt vo mi trng nui cyBm cht kho st v cht chun vo so snh kh nng kh khun

1.4. Mt s v d in hnh c tnh khng khun caoRhazya StrictaL kh ca cy ny c s dng trong vic cha lnh vt thng

Blepharis ciliarisTrong iu tr vt thng v ri lon thn1.4. Mt s v d in hnh c tnh khng khun cao

Francoeuria crispaHt khi ca n iu tr cc chng vim nhim pha trn ngc. S dng c cy1.4. Mt s v d in hnh c tnh khng khun cao

2.1 S lcMalaysia l mt trong nhng quc gia c a dng sinh hc, c bit l h thc vt. Hn 20.000 loi thc vt hoang d c tm thy trong c hn 2000 loi thc vt c dc tnh ang c s dng trong iu tr bnh. Cc hp cht khng vi sinh vt bt ngun t thc vt c tc dng chng li cc vi khun v c tc dng ph t hn so vi thuc khng sinh tng hp. Cc cao th thng c chit vi cc dung mi ph bin nht tch l nc ct, nc kh ion, nc nng, Methanol, Ethyl acetate, Hexane, Chloroform, Dichloro methane, Acetone, ethanol, du th v hn hp Chloroform v Methanol (3: 1).

2. Malaysia

2.2. Phng php sng lc khng khun.a giy lc Whatman c t trong a petri kh.Cho dung dch gc vo a giy lc kh trng.a giy lc c sy kh v t cn thn vo vng nui cy, 37oCCho cht khng sinh chun vo (Nystatin v ciproflaxacin).Quan st tnh khng khun khng nm ca cht th v cht chunPhng php khuch tn a

2.2. Phng php sng lc khng khun.Cho thch nng chy vo a petri to mi trngTri u chng nui cy v trng trn b mtCho 4 dch chit vi nng khc nhau v 1 dung dch chun vo cc ging 37oC trong thi gian ph hpSo snh cc vng khng khunKhuch tn ging thch

2.3. Mt s v d in hnh c tnh khng khun caoAndrographis paniculataNo khang st thng han, khang khun, khang nm, bao v gan, cht chng oxy hoa, khang sinh, chng st ret, khang vim, khang noc rn, khang ung th.Khng c nhiu loi vi khun nh:Proteus mirabilis Proteus vulgaris Neisseria meningitis Escherichia coli Pseudomona saeruginosa

Alpinia conchigera- Thn r c s dng nh mt loi gia v v cc chi non c thm vo mn n t thc vt.Trong y hc dn gian n c s dng iu tr cc vn v tiu ha, vng da, nhim nm, au d dy v nh mt loi thuc sau sinh, chng au, khng vim v khng ung thKhng c nhiu loi vi khun nh:Candida albicansMicrosporum canis Trichophyton rubrum2.3. Mt s v d in hnh c tnh khng khun cao

Euphorbia hirta- N c hot tnh gii lo u, chng oxy ha, chng ung th, khng khun, khng nm, chng st rt, h huyt p, gim au, h st, khng vim .- iu tr cc ri lon n gii, bnh ng h hp nh ho, hen suyn, vim ph qun, ri lon v da, ri lon tiu ha, c bit l k sinh trng ng rut, kit l amoebic, tiu chy v lot.Khng c nhiu loi vi khun nh:Proteus mirablis Proteus vulgaris Proteus aeurginosa

2.3. Mt s v d in hnh c tnh khng khun cao

3. BRAZIL3.1. S lcCerrado l qun x sinh vt ln th hai ca Brazil, sau Amazon, bao gm hn 7.000 loi thc vt c mch. Kho st hot tnh khng khun, khng nm t 5 dch chit khc nhau ca 6 cy thuc t vng Cerrado- min trung Brazil

3.2. Phng php tch chit

nh gi hot tnh khng sinhHot ng khng khun,khng nm c xc nh nh nhau bng th nghim pha long trong mi trng lng trn phin 96 ging ( a c 96 ging).Ghi nhn nng c ch ti thiu (MICg/ml).Th nghim c tnh bng vic xc nh kh nng gy cht trn u trng Brine Shrimp (Artemiasalina).Thc hin trong ba ln. Gi tr LD50 ug/ml c xc nh bng vic phn tch ng tin cy.

3.3. Phng php th hot tnh

Simarouba versicolor iu tr ri lon mu khng c hiu, nhim khun v vim bung trng, t cung.3.4. Mt s v d in hnh c tnh khng khun cao

Roupala brasiliensisiu tr ri lon mu khng c hiu, ng rut

3.4. Mt s v d in hnh c tnh khng khun cao

Hyptis crenataiu tr ri lon h hp, ng rut, bao t3.4. Mt s v d in hnh c tnh khng khun cao

4. n 4.1. S lcHot tnh khng khun ca mt s cy thuc c truyn-dn n cng ang c nghin cu nhiu

4.2. Quy trnh tch chitNguyn liu thc vt c kh di bng mt v nghin thnh bt mn50g bt kh c ngm trong 300ml hexane sut 48 ting kt hp vi lc lin tcCht chit xut t thc vt c lc qua giy lc. Nc lc c sy kh cho n trng lng khng i ta thu c cao chitPhn thc vt cn li c sy kh v ngm trong 300ml methanol v phn on ly trch tin hnh nh quy trnh ly trch bng hexan.

4.3. Chun b mu bnh phm- Nui cy gc c duy tr 4C trn cht dinh dng cha trong thch.- Hot tnh ca vi khun nui cy cho th nghim c to ra bng cch chuyn mt on loopful t t bo ca vt liu nui cy gc n ng nghim ca Mueller-Hinton (MHB) cho vi khun v Sabouraud Dextrose (SDB) cho nm c bit lp trong 24 ting ln lt ti 37C v 25C. - Cc vi khun nui cy c pha long vi MHB v SDB t c mt quang tng ng vi 2,0.106 CFU / ml cho vi khun v 2.0.105 bo t / ml cho nm.

4.4. Th nghim khng sinhRt 15 ml thch nng chy vo a petri v trnga ch cho ng c trong 5 pht v thm vo 0.1% cht nui cy huyn ph c lm ng nht v cht tim chng c lm kh trong 5 phtNng khc ca chit phn on (1.25; 2.5 v 5mg/a) c t trn a v trng 6 mm.a cht c t vo trn b mt thit b v hp cht c php khuch tn trong 5 pht v a c gi cho bnh ti 37C cho 24 ting ng h.Khu vc c ch to thnh quanh a c o bng thc chuyn dng v cch thc tng t c tin hnh tip tc cho nm cn li. Cc th nghim ny c thc hin thnh ba bn.

Trong s 18 loi cy kim tra hot ng chng vi trng, 10 loi thc vt cho thy hot ng khng khun bng cch c ch mt hoc nhiu vi sinh vt. Chit xut Methanol th hin hot tnh khng sinh cao hn so vi chit xut hexane.

4.5. Mt s v d in hnh c tnh khng khun caoToddalia asiatica Pers. SolanaceaeHng liu -thuc b, thuc kch thch, v cay, iu tr bao t.

Punica granatum L. PunicaceaeCht lm se, thuc tr giun sn, kch thch tnh dc, nhun trng, li tiu, thuc v bao t, tim mch, thuc b, cht lm lnh4.5. Mt s v d in hnh c tnh khng khun cao

Cassia fistula L. CaesalpiniaceaeThuc tr giun sn, li tiu, nhun trng, thuc long m, khng vim, thuc x.4.5. Mt s v d in hnh c tnh khng khun cao

5. Nam Phi5.1. S lcC khong 3000 loi thc vt c s dng lm thuc Nam Phi. Trong , c khong 350 loi c s dng ph bin nht. Trong nghin cu ny 12 loi c la chn kim tra hot tnh khng khun chng li tm loi vi sinh vt bao gm nm, vi khun.

5.2. Phng php tch chitCy c phi kh v nghin thnh bt.Ly mi loi bt cy ngm trong Methanol khong 48h nhit phng, khuy trn.LcC quay p sut thp. Thu c cao th.Bo qun 4oC cho n khi cn s dng.

5.3. Phng php th hot tnhMIC, MBC v vi khun c chun b trong microplates 96 gingCc cao th c ha tan bng Dimethylsulfoxide vi nng thch hpCiprofloxacin v M. smegmatis c bm vo ging lm cht chun Mi trng nui cy trn microplates c nim phong vi parafilmQuan st v so mu th vi mu chun

5.3. Phng php th hot tnh Kt qu ca vic xc nh MIC cho thy cc cht chit xut t A. Karoo, E. lasianthum v S. africana c th ngn chn s pht trin ca tt c cc vi sinh vt trong th nghim. Tt c cc mu khc cho thy cc hot ng chn lc.

Acacia karroo HayneLot ming, nm ming, tiu chy, au bng, cm lnh. Bnh hen suyn, vim ph qun, ho, st, bnh phong v bnh h hp.5.4. Mt s v d in hnh c tnh khng khun cao

Erythrophleum lasianthum CorbishleyNhc u, st, bnh tim, vim da, vt thng, bnh thp khp, bnh giang mai, bnh lu, bnh phong, bnh lao, vim ph qun, au tht ngc, v c ca th rng.5.4. Mt s v d in hnh c tnh khng khun cao

Salvia africana-lutea LCm lnh, cm cm, vim ph qun, ho, au tht ngc, ra m hi m, bnh ng h hp v phi.5.4. Mt s v d in hnh c tnh khng khun cao

Khng sinh c ch s pht trin ca vi khun, gi l khng sinh km khun; khng sinh hu hoi vnh vin c vi khun gi l khng sinh dit khun. Tc dng km khun v dit khun thng ph thuc vo nng .

*Khng th c ch c s sinh trng ca nhng vi khun c bit c hi cho con ngi. Do vy, s pht minh mc d rt l th nhng li khng c coi trng.

*ng cha c nng lc v k thut chit xut Penicilline

*Bn cnh cc khng sinh c chit xut t vi sinh vt, cc cht tng hp, bn tng hp, th cc loi thc vt cng sn sinh ra nhng hp cht c kh nng khng khun rt mnh. l ngun nguyn liu di do cho cc nghin cu v hp cht khng khun mi.

*(Catechol c 2 nhm OH cn pyrogallol c 3 nhm OH. V tr v s lng ca cc nhm hydroxyl trn vng phenol c lin quan mt thit n c tnh ca hot cht i vi vi sinh vt. Ngoi ra, ngi ta cn thy rng, phenol trng thi oxy ha cng cao th kh nng c ch vi sinh vt cng tng.)

*Cht juglon (= 5 - hydroxy 1, 4 - naphtoquinon) c trong qu xanhBn cnh juglon cn tn ti dng kh:-hydrojuglon-4--D-glucosid. Qu cn cha mt flavonoid l juglanin.

*y l nhng loi c c cc l mc thnh hoa th gc, trn l c ph lng tuyn bt nhng su b nh b. Cc loiDroseratrn c cha plumbagin v droseron (=2-methyl-3, 5 - dihydroxy -1, 4 - naphtoquinon).Droseron c tc dng lnMycobacterium tuberculosisvi nng 10- 40mg/ml. Drosera cn c tc dng chng co tht do c bo ch di cc dng ru thuc, xir, thuc hm, thuc cao cha ho g v cha ho. ta c cy gng v -D. indicaL. v cy bo t (= c t g) -D. burmanniVahl. c th nghin cu dng lm thuc cha ho.

*Th nghim thuc da.Ly mt ming "da sng ch sn" ngm vo dung dch acid hydrochloric 2% ri ra vi nc ct, sau th vo dung dch nh th trong 5 pht. Ra li vi nc ct ri nhng vo dung dch st (II) sulfat 1%. Nu ming da c mu nu hoc mu nu en l c tanin. " Da sng ch sn" y l mt mng ch t rut ca b v ng vai tr tng t nh da cha thuc.*Trong cy, alkaloid tp trung mt s b phn nht nh. V d, alkaloid tp trung ht nh m tin, c ph, ti c, qu nh t, h tiu, thuc phin, l nh benladon, coca, thuc l, tr, hoa nh c c dc; thn nh ma hong, v nh cankia, mc hoa trng, hong b; r nh ba gc, lu; c nh u, bnh vi, bch b,

**