Kev Tshawb Fawb Txog Kev Tiv Thaiv Ntawm Tag Nrho Glycosides Ntawm Cistanche Deserticola Rau Kev Cuam Tshuam Cuam Tshuam Ntawm Thawj Kab Mob Kab Mob Hepatocytes

Lub hom phiaj: Kawm txog kev tiv thaiv ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm thawj kab mob hepatocytes. Txoj kev: Thawj cov hepatocytes tau sau los ntawm txoj kev hauv situ perfusion, thiab cov txiaj ntsig ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm cov ciaj sia taus ntawm thawj kab mob hepatocytes raug mob los ntawm cawv raug soj ntsuam los ntawm MTT txoj kev; Cov nyhuv ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm morphology ntawm thawj kab lis kev cai hepatocytes thiab nuclei raug mob los ntawm cawv raug soj ntsuam los ntawm fluorescence microscopy; Cov txiaj ntsig ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm apoptosis ntawm thawj kab lis kev cai hepatocytes raug mob los ntawm cawv tau kuaj pom los ntawm kev ntws cytometry; Immunocytochemical staining tau siv los txheeb xyuas cov txiaj ntsig ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm kev qhia ntawm bcl-2 thiab c-fos. Cov txiaj ntsig: Tom qab haus cawv raug mob, qhov ciaj sia ntawm thawj hepatocytes txo qis, thiab muaj kev hloov pauv hauv apoptosis thiab necrosis. Qhov kev qhia ntawm apoptosis-inhibiting gene bcl -2 txo qis, thiab kev qhia ntawm apoptosis-txhim kho gene c-fos nce. Tag nrho cov glycosides ntawm Cistanche deserticola tuaj yeem txhim kho cell ciaj sia tus nqi, txhim kho apoptosis thiab necrosis, txhim khu kev qhia ntawm bcl -2, thiab inhibit qhov kev qhia ntawm c-fos. Cov nyhuv yog nyob ntawm koob tshuaj. Xaus: Cistanche deserticola glycosides tuaj yeem tiv thaiv thawj kab lis kev cai hepatocytes los ntawm kev nthuav tawm cov kab mob apoptosis-inhibiting gene bcl-2, txo qhov kev qhia ntawm apoptosis-txhim kho cov noob c-fos, txo apoptosis thiab necrosis, thiab nce cell ciaj sia taus.

Ntsiab lustag nrho glycosides ntawmcistanche deserticola; Thawj kab lis kev cai; Hepatocyte; Alcoholic daim siab raug mob; Apoptosis

Cistanche hmoov

Nyob rau hauv xyoo tas los no, nrog kev txhim kho ntawm tib neeg txoj kev ua neej nyob, kev haus cawv tau nce sai, nrog rau kev nce ntau yam kab mob cawv, thiab haus cawv rau lub siab yog ib qho teeb meem tseem ceeb. Cov kev tshawb fawb tau pom tias Cistanche deserticola muaj kev tiv thaiv los ntawm kev txhim kho kev tiv thaiv kab mob, tiv thaiv kev laus, tiv thaiv hluav taws xob, tiv thaiv oxidation, tiv thaiv lipid peroxidation, thiab cawv ua rau lub siab raug mob [1]. Glycosides yog cov khoom tseem ceeb ntawm Cistanche deserticola.Hauv kev sim yav dhau los, nws tau txheeb xyuas los ntawm kev sim tsiaj uas tag nrho glycosides ntawmCistanche deserticola maNws tuaj yeem txo qis kev tsim cov dawb radicals, txhim kho cov peev txheej ntawm cov dawb radicals thiab lawv cov metabolites, yog li inhibiting lipid peroxidation, thiab ua lub luag haujlwm tiv thaiv kab mob hauv siab ua rau lub siab raug mob.[2]. Qhov kev sim no yuav tshawb txog kev tiv thaiv ntawm tag nrho cov glycosides ntawm Cistanche deserticola ntawm cov dej cawv daim siab raug mob ntawm qib cellular los ntawm thawj kab lis kev cai ntawm hepatocytes.

1 Cov khoom siv thiab cov txheej txheem

1.1 Reagent thiab ntsuasTag nrho glycosides ntawmCistanche deserticola, tsaus xim av hmoov, muab los ntawm Lub Tsev Kawm Ntawv ntawm Pharmacy ntawm Lanzhou University, muaj 88.6 feem pua ​​​​ntawm tag nrho glycosides ntawm phenyl ethanol. Dissolve nyob rau hauv ob chav distilled dej thiab dilute cov kab lis kev cai tov rau qhov yuav tsum tau concentration. Enzyme marker (Beckman Coulter AD340), fluorescence microscope (Olympus, BX51), inverted phase contrast microscope (Leica DMI3000B), flow cytometry (Beckman colter cell, CellLabQuanta SC), thiab lwm yam.

Cistanche hmoov

Nyem qhov no mus saib Cistanche deserticolativ thaiv daim siabcov khoom

【Nug ntxiv】 Email: xue122522@foxmail.com /  Whats App:  0086 18599088692 /  Wechat:  18599088692

1.2 Cov thawj hepatocytes raug cais tawm thiab coj los ntawm txoj kev hauv situ perfusion [3].

Ib tus nas Kunming tau tshuaj loog los ntawm {{0}}.5 feem pua ​​pentobarbital 0.5 mL intraperitoneal txhaj. Tom qab niaj hnub tshuaj tua kab mob ntawm daim tawv nqaij, qhib lub plab, txav lub plab zom mov mus rau sab laug, nthuav tawm cov hlab ntsha portal, ua tib zoo puncture koob infusion ntawm cov cuab yeej infusion los ntawm qhov kawg ntawm lub portal leeg, thiab ligate thiab kho nws. Tig cov dej ntws ntawm qhov dej tso rau qhov siab tshaj plaws (kwv yees li 4mL / min), thiab tso cov tshuaj calcium uas tsis muaj dej ua ntej preheated ntawm 37 degree. Thaum lub siab o, txiav tawm qhov kawg ntawm qhov kawg ntawm qhov inferior vena cava sai sai, thiab compress lub proximal kawg ntawm inferior vena cava intermittently kom lub siab alternately retracts thiab expands. Thaum cov ntshav seem raug ntxuav tawm thiab lub siab yog xim daj thiab dawb, qhov sov so 0.05 feem pua ​​collagenase (Sigma) perfusion tov yog siv rau kev zom zaub mov. Kev zom zaub mov nres thaum daim siab ua mos thiab inelastic, me vacuoles tshwm nyob rau hauv daim siab capsule, los yog tej kab nrib pleb tshwm nyob rau hauv daim siab cov ntaub so ntswg. Ua tib zoo dissociate lub siab, hloov mus rau ib lub phaj tsis muaj menyuam uas muaj qhov tsim nyog ntawm DMEM high-qab zib kab lis kev cai nruab nrab (Sigma), tev tawm lub siab lub hnab ntawv thiab maj mam co nws, ces tawg cov hepatocytes nyob rau hauv kab lis kev cai nruab nrab, sau lawv nyob rau hauv ib tug conical. raj, co thiab kab lis kev cai rau 10 min (kev vibration zaus yog 100 zaug / min), centrifuge rau 3 min (4 degree , 1000r / min), thiab pov tseg lub supernatant. Cov hlwb raug tshem tawm nrog cov kab lis kev cai nruab nrab, lim nrog 200 mesh nylon mesh, thiab cov filtrate tau centrifuged ntau zaus 3 zaug (4 degree, 1000 r / min) kom tau lub siab parenchyma hlwb. Kho cov cell concentration mus rau 5 × Tom qab 106 hlwb / mL, inoculate lawv mus rau hauv 25mL kab lis kev cai hwj pretreated nrog nas tail collagen, thiab ces incubate lawv nyob rau hauv ib tug 37 degree, 5 feem pua ​​CO2 incubator. Tom qab 24 teev, hloov cov kev daws teeb meem, pov tseg cov hlwb uas tsis yog-adherent, thiab txuas ntxiv mus rau kab lis kev cai rau kev siv hauv kev sim tom ntej.

1.3 Kev txiav txim siab ntawm kev loj hlob nkhaus ntawm thawj kab mob nas hepatocytes

Kho cov hepatocytes purified mus rau 5 × 106 hlwb/mL tau inoculated rau ntawm 24-zoo phaj, thiab 1 mL ntawm cell suspension twb inoculated nyob rau hauv txhua lub qhov dej. Tag nrho ntawm 21 lub qhov dej tau raug inoculated, thiab DMEM high-qab zib kab lis kev cai nruab nrab uas muaj 10 feem pua ​​​​fetal bovine serum (Sigma) tau coj mus kuaj rau 7 hnub. Noj 3 qhov ntawm random txhua txhua hnub, zom thiab tshuab 0.25 feem pua ​​​​ntawm trypsin tusyees, suav rau ntawm lub rooj tsavxwm suav, thiab kos qhov kev loj hlob nkhaus

1.4 Tsim cov qauv kev raug mob ntawm cov kab mob cawv hepatocyte.

Cov logarithmic hepatocytes tau los ntawm txoj kev saum toj no tau inoculated rau hauv {{{0}}} zoo phaj pretreated nrog nas tail collagen, thiab 100 hlwb raug inoculated rau hauv txhua lub qhov dej μ 50. DMEM kab lis kev cai nruab nrab ntawm 10 feem pua ​​​​fetal bovine serum tau kab lis kev cai rau 24 teev thiab tom qab ntawd hloov mus rau lwm 24 teev. Ib pawg dawb huv tau teeb tsa, thiab tsib pawg kev cuam tshuam cawv (50, 75, 100, 125, 150 mmol / L) tau teeb tsa, thiab txhua pab pawg tau teeb tsa nrog tsib qhov rov ua dua. Tom qab cov kab mob hepatocytes tau coj los ntawm ntau qhov sib txawv ntawm cawv rau 24h, 0.5 feem pua ​​​​MTT (Sigma) 20 tau ntxiv rau txhua lub qhov dej μ 50. Txuas ntxiv mus rau kab lis kev cai rau 4h, pov tseg cov supernatant, thiab ntxiv DMSO150 rau txhua lub qhov dej μ 50. Muab tso rau hauv ib lub qhov dej. co lub rooj thiab sib tov rau 15min, ntsuas tus nqi absorbance ntawm 570nm, thiab xam cov cell ciaj sia taus. Cell ciaj sia tus nqi ntawm pawg tswj hwm / tswj pawg × 100 feem pua

1.5 Cov txiaj ntsig ntawm tag nrho glycosides ntawm cistanche deserticola ntawm kev ciaj sia ntawm thawj kab mob hepatocytes

Kev raug mob los ntawm cawv Inoculate lub hepatocytes tau los ntawm cov saum toj no txoj kev ntawm lub {{{0}} zoo phaj pretreated nrog nas tail collagen thiab inoculate txhua lub qhov dej nrog 100 μ 50. Lub DMEM kab lis kev cai nruab nrab ntawm 10 feem pua ​​​​fetal bovine serum tau coj mus kuaj rau 24 teev thiab tom qab ntawd hloov. Tom qab lwm 24 teev, GCs nrog qhov kawg siab ntawm 0.2, 0.4, thiab 0.8 g / L tau ntxiv rau pawg tswj hwm, raws li. Pawg qauv ethanol thiab pab pawg dawb huv txuas ntxiv mus rau kab lis kev cai yam tsis muaj tshuaj ntxiv, thiab txhua pab pawg tau teeb tsa nrog tsib lub qhov dej ntau. Tom qab 24 teev, pawg qauv thiab pawg GCs cuam tshuam tau ntxiv nrog cawv nrog qhov kawg siab ntawm 100 mmol / L. Tom qab 12 teev ntawm kab lis kev cai, lub cell ciaj sia taus yog ntsuas los ntawm MTT txoj kev.

1.6 Cov txiaj ntsig ntawm tag nrho glycosides ntawm cistanche deserticola ntawm morphology ntawm thawj kab lis kev cai hepatocytes thiab nuclei raug mob los ntawm cawv

Muab cov iav npog ua ntej ua kom tsis muaj menyuam rau hauv 6- zoo phaj nrog qhov inoculation concentration ntawm 5 × 106 cells/mL cell suspension, qauv pawg, Cistanche deserticola pawg (0.2 , 0.4, {{20}}.8 g/L) thiab pab pawg dawb huv. Tom qab 24 teev ntawm kab lis kev cai, ntxiv 100 mmol / L cawv nrog qhov kawg concentration. Tom qab 24 teev ntawm kab lis kev cai, tshem tawm 95 feem pua ​​​​ethanol los ntawm iav swb thiab kho rau 15 feeb. PBS tau ntxuav ob zaug, thiab tom qab ntawd rov faib rau hauv PBS. Kho cov cell concentration rau 5 × 104 hlwb / mL, ntxiv 10 μ L ntawm acridine txiv kab ntxwv - ethylidene bromide sib tov (Sigma) (0.01 μ G / mL acridine txiv kab ntxwv kua, 0.02 μ G / mL promethazine tov, tov ntim ratio: 1/ 1), incubated rau 30min, soj ntsuam thiab yees duab nyob rau hauv lub fluorescence microscope.

1.7 Cov nyhuv ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm apoptosis ntawm thawj kab lis kev cai hepatocytes raug mob los ntawm cawv

Kho qhov ceev ntawm digested cell suspension mus rau 5 × 106 hlwb/mL, inoculated nyob rau hauv ib tug rau-zoo phaj thiab kab lis kev cai rau 24 teev, teem qauv pab pawg neeg, cistanche deserticola pawg (0.2 , 0.4, 0.8 g/L) thiab pab pawg dawb. Tom qab 24 teev ntawm kab lis kev cai, ntxiv cawv nrog qhov kawg ntawm 100 mmol / L, incubate rau 24 teev, zom thiab sau cov hlwb, ntxuav lawv nrog PBS ob zaug, kho lawv ntawm 4 degree ib hmos nrog 70 feem pua ​​​​precooled ethanol, ntxuav lawv nrog PBS. tov ob zaug, ntxiv RNA enzyme (Sigma), PI (100 μ G / mL) (Sigma), thiab muab tso rau hauv 37 degree dej da dej rau 30min. Tus nqi apoptosis tau ntsuas los ntawm kev ntws cytometry.

1.8 Tag nrho glycosides ntawm Cistanche deserticola tuaj yeem cuam tshuam cov apoptosis ntawm thawj kab mob hepatocytes raug mob los ntawm cawv, bcl-2, thiab pro-apoptotic gene c-fos

Cov txiaj ntsig ntawm fos qhia tau raug coj los ntawm peb daim phiaj rau rau kab lis kev cai, txhua tus yog ib pawg ntawm peb qhov dej. Pawg qauv, pawg Cistanche deserticola (0.2, 0.4, 0.8 g/L), thiab pawg dawb huv tau teeb tsa. Ib qho sterile swb tau muab tso rau hauv txhua lub qhov. Lub logarithmic kev loj hlob theem hlwb raug coj mus, thiab trypsin digestion yog siv los npaj ib leeg cell suspension, thiab cell ceev tau hloov mus rau 5 × 106 / mL thiab inoculated rau ntawm ib tug 6-zoo phaj, 2mL / zoo. Tom qab 24 teev ntawm kab lis kev cai, ntxiv cawv nrog qhov kawg ntawm 100 mmol / L, thiab txuas ntxiv mus rau kab lis kev cai rau 24 teev. Tshem tawm cov swb puv ntawm cov hlwb, ntxuav lawv nrog PBS ob zaug, thiab zas lawv nrog cov txheej txheem immunohistochemical ABC niaj hnub. Cov kauj ruam luv luv yog raws li hauv qab no: kho lawv nrog 40 feem pua ​​​​ntawm paraformaldehyde rau 10 feeb, kaw lawv nrog cov yaj cov ntshav rau 30 feeb, poob thiab ntxiv bcl -2 polyclonal antibody (Invitrogen) (1:75) lossis c-fos polyclonal antibody (Invitrogen) (1:50), react nyob rau hauv chav tsev kub rau 1.5 teev, ntxiv ABC complex ntawm chav tsev kub rau 2 teev, xim DAB, thiab muab lawv nrog gelatin. Qhov kev qhia ntawm Bcl-2 thiab c-fos protein tau muab piv los ntawm lub teeb tsom iav thiab thaij duab.

1.9 Kev ua lej Cov ntaub ntawv sim tau ua tiav nrog SPSS13.5 software. Nws yog qhia raws li qhov txhais tau tias ± tus qauv sib txawv (x ± s), thiab t-test yog siv rau kev sib piv ntawm pab pawg. P<0.05 indicates that the difference is statistically significant.

2 Kev tshwm sim

2.1 Txoj kev loj hlob ntawm thawj kab lis kev cai nas hepatocytes tau pom nyob rau hauv daim duab 1 Nws tuaj yeem pom tias cov hepatocytes nce ntxiv nrog lub sijhawm incubation, thiab nkag mus rau theem kev loj hlob ntawm logarithmic rau hnub 4.

Fig. 1 Kev loj hlob nkhaus ntawm thawj kab mob nas hepatocytes

2.2 Cov txiaj ntsig ntawm qhov sib txawv ntawm cov cawv ntawm kev muaj sia nyob ntawm thawj kab mob nas hepatocytes

Pom tau los ntawm Table 1 tias cawv muaj kev puas tsuaj rau thawj kab mob hepatocytes, thiab cov nyhuv yog nyob ntawm koob tshuaj. Raws li qhov xav tau ntawm qhov kev sim, 100mmol / L yuav raug siv los ua kev ua haujlwm siab hauv qhov kev sim tom ntej, thiab cov cell ciaj sia taus yog 43.40 feem pua.

2.3 Cov nyhuv ntawm tag nrho glycosides ntawmcistanche deserticolantawm cov ciaj sia taus ntawm thawj kab lis kev cai hepatocytes raug mob los ntawm cawv

Muaj peev xwm pom tau nyob rau hauv Table 2. Tom qab haus cawv raug mob, cov ciaj sia ntawm cov hepatocytes yuav txo qis. Tag nrho cov glycosides ntawm cistanche deserticola tuaj yeem ua rau muaj sia nyob ntawm hepatocytes (P<0.05), and the effect is dose-dependent (P<0.05).

Suab puam nyob cistanche

2.4 Cov nyhuv ntawm tag nrho glycosides ntawmcistanche deserticolantawm morphology ntawm thawj kab lis kev cai hepatocytes thiab nuclei puas los ntawm cawv. 

Azidine txiv kab ntxwv qhia ntsuab fluorescence tom qab nkag mus rau hauv lub hlwb, thaum promethazine tsuas stain lub necrotic hlwb nrog ib tug tsis tiav cell membrane. Nws tau pom los ntawm kev soj ntsuam (Daim duab 2) tias cov hepatocytes nyob rau hauv cov pab pawg neeg dawb paug yog uniformly ntsuab, thiab tsis muaj bromine liab staining ntawm necrotic hlwb (A); Tom qab kev kho mob nrog cawv, cov hepatocytes tau me me, thiab ntau lub hlwb necrotic stained nrog bromine liab thiab apoptotic hlwb nrog chromatin concentration nyob rau hauv lub nucleus (lub nucleus yog ci ntsuab) (B) tshwm sim. Yog li, nws ntseeg tau tias cawv kuj tuaj yeem ua rau cell necrosis thaum inducing hepatocyte apoptosis. GCs tuaj yeem txo qis necrosis thiab apoptosis, thiab cov txiaj ntsig zoo sib xws nrog cov koob tshuaj (C, D, E).

Daim duab 2 Kev soj ntsuam cov txiaj ntsig ntawm tag nrho cov glycosides ntawm cistanche deserticola ntawm morphology ntawm thawj kab mob siab lub siab nuclei nyob rau hauv fluorescence microscope (acridine txiv kab ntxwv staining, 400)

A: Pab pawg dawb; B: Qauv pawg; C:{{0}}.2g/L-GCs; D:0.4g/L-GCs; E: 0.8g / L-GCs

2.5 Cov nyhuv ntawm tag nrho glycosides ntawmcistanche deserticolantawm apoptosis ntawm thawj kab lis kev cai hepatocytes raug mob los ntawm cawv

Cistanche extract hmoov

【Nug ntxiv】 Email: xue122522@foxmail.com /  Whats App:  0086 18599088692 /  Wechat:  18599088692

tuaj yeem pom hauv daim duab 3-B, tus nqi apoptosis ntawm hepatocytes tom qab haus cawv raug mob tau nce ntau (34.7 feem pua), thiab tag nrho cov glycosides ntawm cistanche deserticola tuaj yeem ua rau muaj kev vam meej ntawm hepatocytes hauv kev noj tshuaj, uas yog raws li cov txiaj ntsig tau pom nyob rau hauv fluorescence microscope.

Fig. 3 Cov nyhuv ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm apoptosis ntawm thawj kab lis kev cai hepatocytes raug mob los ntawm cawv

A: Pab pawg dawb; B: Qauv pawg; C:{{0}}.2g/L-GCs; D:0.4g/L-GCs; E: 0.8g / L-GCs

Los ntawm immunocytochemistry, nws tau pom tias qhov kev qhia ntawm apoptosis-inhibiting factor bcl-2 txo qis (4-B) thiab kev qhia ntawm c-fos nce (5-B) tom qab lub siab hlwb raug mob. ntawm cawv. Tag nrho glycosides ntawm Cistanche deserticola tuaj yeem txhim kho qhov kev qhia ntawm bcl-2 (4-C, D, E) thiab inhibit qhov kev qhia ntawm c-fos (5-C, D, E). Cov txiaj ntsig tau pom hauv daim duab. 4-5.

Fig. 4 Cov nyhuv ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm kev qhia ntawm bcl-2 hauv thawj kab lis kev cai hepatocytes raug mob los ntawm cawv (400)

A: Pab pawg dawb; B: Qauv pawg; C:{{0}}.2g/L-GCs; D:0.4g/L-GCs; E: 0.8g / L-GCs

Fig. 5 Cov nyhuv ntawm tag nrho glycosides ntawm Cistanche deserticola ntawm kev qhia ntawm c-fos hauv thawj kab lis kev cai hepatocytes raug mob los ntawm cawv (400)

A: Pab pawg dawb; B: Qauv pawg; C:{{0}}.2g/L-GCs; D:0.4g/L-GCs; E: 0.8g / L-GCs

3 Kev sib tham

Suav tshuaj ntsuab cistanche

Berry et al. thawj zaug tsim cov txheej txheem in-situ perfusion ntawm kev siv collagenase kom perfuse lub siab kom tau hepatocytes nyob rau hauv physiological mob, thiab tom qab ntawd tsim los ntawm Seglen li al. [4] ua kom lub tshuab collagenase perfusion no zoo meej dua. Txoj kev no muaj cov cell yield siab thiab muaj sia nyob siab. Hauv qhov kev sim no, thawj cov kab mob hepatocytes tau txais los ntawm kev sib xyaw ntawm nas hepatocytes thiab kab lis kev cai. Txoj kev tau ruaj khov thiab kev ua haujlwm ntawm tes muaj zog. Los ntawm kev ntsuas kev loj hlob nkhaus, nws tau pom tias thawj hepatocytes tau nkag mus rau theem kev loj hlob ntawm logarithmic tom qab 4 hnub.

MTT colorimetric microanalysis yog ib txoj hauv kev rhiab heev los kuaj xyuas cov cell loj hlob thiab muaj sia nyob nrog kev rov ua tau zoo. Succinate dehydrogenase nyob rau hauv mitochondria ntawm cov hlwb muaj sia tuaj yeem txo cov daj MTT mus rau xiav-violet crystals, thaum lub hlwb tuag tsis muaj haujlwm zoo li no [5]. Kev ua haujlwm ntawm tes tuaj yeem cuam tshuam los ntawm kev ntsuas qhov muag pom. Hauv qhov kev sim no, MTT txoj kev tau siv los kuaj pom tias qhov muaj sia nyob ntawm cov hepatocytes tom qab haus cawv raug txo qis, thiab tag nrho glycosides ntawm Cistanche deserticola tuaj yeem ua rau kom muaj sia nyob ntawm hepatocytes (P.<0.05), and the effect was dose-dependent (P<0.05).

Apoptosis yog ib hom tshwj xeeb ntawm kev tuag ntawm tes. Thaum cov hlwb raug apoptosis, tshwj xeeb morphological thiab biochemical hloov pauv yuav tshwm sim [6-8]. Qhov kev sim tau lees paub tias feem ntau ntawm cov hepatocytes yog apoptotic tom qab kev kho dej cawv, thiab cov hlwb tau pom cov kev hloov pauv ntawm morphological, uas yog raws li hauv qab no: lub hlwb ua me me, microvilli ntawm lub xov tooj ntawm tes ploj mus, lub xov tooj ntawm tes poob qis, muaj vacuoles, nuclear. condensation, thiab tsim ntawm apoptotic lub cev; Nws yog nrog los ntawm necrosis. Tom qab kev kho mob nrog tag nrho glycosides ntawm Cistanche deserticola, tus nqi apoptosis poob qis, thiab qhov teeb meem necrosis kuj tau yooj yim.

bcl -2 noob yog proto-oncogene. Tam sim no, tsib bcl-2 noob tsev neeg (bcl-2, bcl-2 x, bax, mcl-1, thiab A1) tau pom, uas bcl{{7 }} tuaj yeem inhibit programmed cell tuag (PCD). Kev ua haujlwm ntawm bcl-2Cov txheej txheem yuav suav nrog: (1) inhibiting tso tawm Ca2 ntxiv . Bcl -2 yog ib qho transmembrane protein, uas feem ntau yog nyob rau ntawm daim nyias nyias. Lub sab hauv thiab sab nraud nuclear membranes txuas nrog lub lumen ntawm endoplasmic reticulum. Lub tom kawg yog qhov chaw khaws cia tseem ceeb ntawm intracellular Ca2 ntxiv, uas ua lub luag haujlwm tseem ceeb hauv cov txheej txheem ntawm cell apoptosis. Siv cov txheej txheem transgenic, nws tau pom tias qhov kev qhia siab ntawm bcl -2 tuaj yeem cuam tshuam kev tso tawm ntawm Ca2 ntxiv los ntawm endoplasmic reticulum, yog li nws tau kwv yees tias cov nyhuv inhibitory ntawm bcl -2 ntawm apoptosis yuav cuam tshuam. rau Ca2 ntxiv rau hauv endoplasmic reticulum [9]. (2) Bcl-2 ua lub luag haujlwm tiv thaiv apoptotic los ntawm kev thaiv cov teeb liab kis ntawm cov noob pro-apoptotic, lossis los ntawm kev thaiv cov khoom lag luam noob tsis muaj zog. Cov kev tshawb fawb tau pom tias bcl -2 protein tuaj yeem tiv thaiv apoptosis tshwm sim los ntawm P53 thiab c-fos [10]. (3) Bcl-2 tuaj yeem ua lub luag haujlwm tiv thaiv apoptotic los ntawm inhibiting dawb radicals. bcl -2 muaj lub luag haujlwm ntawm antioxidant, uas tuaj yeem cuam tshuam kev tsim thiab kev ua ntawm superoxide, yog li inhibiting qhov tshwm sim ntawm apoptosis [11].

Cov noob ntxov ntxov (IEGs) yog cov noob uas tshwm sim ntxov thiab muaj lub sijhawm luv luv nyob rau hauv ntau yam teeb meem stimuli. c-fos yog ib qho ntawm cov proto-oncogenes, nyob ntawm chromosome 14q21-q31. Nws yog 9kb DNA, tsim ntawm plaub exons thiab peb introns [12]. Cov ntawv sau yooj yim tsis tshua muaj, tab sis nws tuaj yeem tshwm sim los ntawm qhov sib txawv ntawm tus tub txib thib ob rau kev nthuav tawm sai thiab hloov pauv. Cov khoom protein ntawm c-fos yog FOS, thiab cov protein encoded los ntawm proto-oncogene c-Jun yog JUN. FOS muaj leucine anti-chain (LZ) [13]. FOS thiab JUN tsim ib qho heterodimer nyob rau hauv lub nucleus los ntawm LZ qauv, hu ua transcription factor AP-1, uas ua ke nrog AP -1 qhov chaw ntawm lub hom phiaj noob thiab ua raws li tus xa xov thib peb ntawm lub nucleus, ua rau kev qhia ntawm cov nyhuv noob rau lub sijhawm ntev [14] Morgan ntseeg tias qhov sib txawv ntawm c-fos qhia txog kev sib txawv thiab qhov nyuaj ntawm nws txoj haujlwm. Cov lus qhia ib ntus tuaj yeem koom nrog hauv kev tiv thaiv ntawm tes, kho, kho dua tshiab, thiab rov tsim dua tshiab, thaum lub sijhawm overexpression muaj feem cuam tshuam rau kev puas tsuaj thib ob. Qhov kev qhia mechanism ntawm ob yog txawv. Yog li, nws tuaj yeem txiav txim siab tias IEGs xws li c-fos ua si ob lub luag haujlwm. Thaum kho kev tiv thaiv kab mob yog tag nrho inhibited, lawv koom nrog apoptosis; Thaum nws tsis yog inhibited, nws muaj kev tiv thaiv ntawm cov hlwb nyob ib ncig ntawm qhov txhab thiab koom nrog hauv cov txheej txheem ntawm kev kho cell thiab rov tsim dua tshiab [15]. WebsterKR siv antisense nucleotides los txo qhov kev hloov pauv ntawm c-fos thiab tiv thaiv qhov tshwm sim ntawm apoptosis, qhia tias qhov overexpression ntawm c-fos yog inextricably cuam tshuam rau qhov tshwm sim thiab kev loj hlob ntawm ntau yam kab mob [16].

Hauv qhov kev sim no, tom qab lub siab hlwb raug mob los ntawm kev haus cawv, kev qhia ntawm apoptosis-inhibiting factor bcl-2 tau txo qis, thiab kev qhia ntawm c-fos tau nce, qhia tias tag nrho glycosides ntawm Cistanche deserticola tuaj yeem tiv thaiv daim siab. hlwb los ntawm kev txhim kho cov kev qhia ntawm bcl -2, inhibiting kev qhia ntawm c-fos, txo lub siab cell raug mob thiab apoptosis.

【Nug ntxiv】 Email: xue122522@foxmail.com /  Whats App:  0086 18599088692 /  Wechat:  18599088692