PART 1 The Phenylethanol Glycoside Liposome Inhibits PDGF-Induced HSC Activation Via Regulation Of FAK/PI3K/Akt Signaling Pathway
Mar 06, 2022
Shi-Lei Zhang 1, Long Ma 1, Jun Zhao 2, Shu-Ping You 1, Xiao-Ting Ma 1, Xiao-Yan Ye thiab Tao Liu 1, *
1 Department of Toxicology, School of Public Health, Xinjiang Medical University, Xinjiang Uyghur Autonomous Region, Xinyi Road No.393, Urumqi 830011, Suav teb
2 Lub Chaw Haujlwm Tseem Ceeb rau Uighur Tshuaj, Lub Tsev Kawm Ntawv ntawm Materia Medica ntawm Xinjiang, Xinjiang Uyghur Autonomous Region, Tianshan District, Xinhua South Road No. 140, Urumqi 830004, Suav
Yog xav paub ntxiv thov hu rau:Joanna.jia@wecistanche.com
Cistanche deserticola muaj ntau yam teebmeem, nyem qhov no kom paub ntau ntxiv
Abstract: Cistanche tubulosayog ib hom tshuaj suav tshuaj ntsuab uas tau siv dav los tswj kev tiv thaiv kab mob, thiabphenylethanoid glycosides(CPhGs) yog ib feem tseem ceeb ntawm cov haujlwm no. Txawm li cas los xij, kev siv CPhGs cuam tshuam tsis zoo los ntawm lawv qhov kev nqus tsis zoo thiab kev siv qhov ncauj tsawg. Lub hom phiaj xa tshuaj yog ib qho tseem ceeb ntawm kev txhim kho cov tshuaj. Cov kev tshawb fawb yav dhau los tau qhia tias CPhGs tuaj yeem thaiv qhov kev coj ua ntawm txoj kev taw qhia hauv TGF-p1 / smad thiab inhibit qhov ua kom lub siab ntawm cov hlwb hlwb (HSCs). Lub hom phiaj ntawm txoj kev tshawb no yog los soj ntsuam cov nyhuv anti-hepatic fibrosis ntawm CPhG liposomes los ntawm inhibiting HSC activation, txhawb kev apoptosis, thaiv lub voj voog ntawm tes, inhibiting kev coj ntawm kev taw qhia hauv focal adhesion kinase (FAK) / phosphatidylinositol-3- kinase (PI3K) / protein kinase B (Akt), thiab txiav txim siab lawv nyob hauv vitro hepatoprotective kev ua haujlwm. Cov kev tshawb fawb hauv vitro tso tawm tau pom tias CPhG liposomes muaj qhov cuam tshuam tso tawm piv rau cov tshuaj CPhGs. HSC proliferation tau inhibited tom qab kho nrog CPhG liposomes (29.45,14.72, 7.36 ^g / mL), nrog IC50 qhov tseem ceeb ntawm 42.54 Rg / mL hauv MTT kev soj ntsuam. Qhov sib txawv ntawm CPhG liposomes tuaj yeem cuam tshuam HSC proliferation, txhawb kev apoptosis, thiab thaiv lub voj voog ntawm tes. MTT txoj kev qhia pom tseeb inhibition ntawm HSC proliferation tom qab CPhG liposome thiab Recombinant Rat Platelet-derived growth factor-BB(rrPDGF-BB) kho. Cov theem ntawm collagen{17}}, metallopeptidase inhibitor 1 (TIMP-1), cov nqaij mos actin (a-SMA), thiab phosphorylated PI3K/Akt tau downregulated, thiab matrix metalloproteinase-1 (MMP{ {23}}) tau tswj hwm, los ntawm kev kho ua ntej nrog ntau qhov sib txawv ntawm CPhG liposomes. Ntxiv mus, 29.45 Rg / mL ntawm CPhG liposomes tuaj yeem txo qhov kev qhia ntawm FAK protein thiab phosphorylated PI3K thiab Akt protein downstream ntawm FAK los ntawm overexpression ntawm FAK noob. Qhov kev sim no qhia tias CPhG liposomes tuaj yeem cuam tshuam qhov ua kom HSCs los ntawm inhibiting FAK thiab tom qab ntawd txo qhov kev qhia ntawm phosphorylated Akt / PI3K, yog li muab kev nkag siab tshiab rau hauv daim ntawv thov ntawm CPhGs rau daim siab fibrosis.
Ntsiab lus: phenylethanoid glycosidesliposome; hepatic stellate hlwb; kev loj hlob; apoptosis; cell voj voog; FAK/PI3K/Akt,
yam, xws li TGF-p1, cawv, co toxins, thiab steatosis, tuaj yeem ua rau HSCs ua kom muaj zog. PDGF tseem tuaj yeem ua rau kev ua kom HSCs [4]. Tam sim no, cov tswv yim kho mob tsom rau HSCs los tiv thaiv daim siab fibrosis muaj xws li inhibition of HSCs activation, proliferation, and cell cycle, as well as the stimulation of HSCs apoptosis [5].
Cistanche tubulosa(tsev neeg Orobanchaceae), ib tsob nroj cab kab mob, tau loj hlob nyob rau sab qab teb ntawm Xinjiang hauv Suav teb.C. tubulosatau pom los nthuav qhia ntau yam dej num, suav nrog kev txhim kho kab mob kev tiv thaiv kab mob, txhim kho kab mob endurance, nourishing lub raum, kho impotence, thiab nce kev txawj ntse;
1. Taw qhia
nws kuj muaj anti-oxidation thiab anti-aging zog [6]. Cov nroj tsuag no muaj phenylethanoid glycosides (CPhGs), iridoids, thiab polysaccharides, uas CPhGs yog ib co ntawm cov tseem ceeb active bioactive hom [7]. Hauv kev tshawb fawb yav dhau los, peb pom tias CPhGs tuaj yeem thaiv qhov kev coj ua ntawm txoj kev taw qhia hauv TGF-p1 / smad, inhibit qhov ua kom HSCs, thiab tiv thaiv cov teebmeem ntawm bovine serum albumin-induced hepatic fibrosis hauv nas [7,8]. Txawm li cas los xij, CPhGs muaj cov dej zoo solubility thiab tsis zoo lipid solubility thiab cuam tshuam los ntawm lwm yam xws li acidity thiab decomposition enzymes, uas txo cov stability thiab kev ua tau zoo ntawm CPhGs [9]. Tom qab kev tswj hwm intragastric, CPhGs tsis ruaj khov nyob rau hauv lub plab zom mov ntawm nas, thiab hydrolysis ntawm glycoside bonds tuaj yeem tshwm sim tau yooj yim. Qhov ncauj bioavailability ntawm nas tsuas yog 0.83 feem pua, yog li nws nyuaj rau txiav txim siab nws txoj haujlwm kho mob [10]. Yog li ntawd, peb yuav tsum nrhiav kev nyab xeeb, ua tau zoo, txhawb nqa, thiab tsom rau cov tshuaj tiv thaiv kab mob rau HSCs txhawm rau txhim kho lawv txoj kev kho mob. Nanoparticle tshuaj xa cov tshuab muab lwm lub tswv yim los kov yeej cov kev tsis txaus no vim lawv qhov zoo, suav nrog cov tshuaj muaj peev xwm thauj khoom siab thiab cov ntshav mus ntev [11]. Lawv muaj kev sib raug zoo rau lub siab thiab tuaj yeem xa cov tshuaj ncaj qha mus rau hauv cov hlwb los ntawm endocytosis thiab fusion. Unmodified liposomes feem ntau yog muab faib rau hauv cov ntaub so ntswg los yog cov kab mob uas tsim los ntawm reticuloendothelial system (RES) thiab muaj kev ua haujlwm ntawm passively targeting lub siab [12]. Yog li ntawd, CPhG liposomes tau npaj los ntawm cov membrane dispersion thiab thib ob encapsulation los muab cov nyhuv anti-fibrosis. Hauv txoj kev tshawb no, peb tsom mus tshawb xyuas qhov cuam tshuam ntawm CPhG liposomes ntawm HSC proliferation, apoptosis, thiab cell voj voog, nrog rau nws cov txheej txheem.
2. Cov txiaj ntsig
2.1. Physical Properties ofCPhG Liposome
Cov khoom ntawm lub cev ntawm CPhG liposomes tau npaj raws li hauv qab no: Qhov loj me yog (216.7 ± 3.47) nm, Zeta muaj peev xwm yog (-55.6 ± 1.3) mV. Raws li pom nyob rau hauv daim duab 1, cov hais yog puag ncig-zoo li, coated nrog ib tug uniform thickness ntawm qhov chaw, thiab particle loj tis yog uniform. Cov tshuaj loading thiab encapsulation efficiency ntawm CPhG liposomes yog (3.71 ± 0.32) feem pua thiab (38.46 士 7.85) feem pua.
2.2. Hauv Vitro Tso Tawm los ntawm CPhG Liposomes thiab CPhGs
Qhov kwv yees kwv yees tso tawm tus nqi tau suav thiab npaj ua lub sijhawm ua haujlwm rau CPhG liposomes thiab CPhGs (Daim duab 2). Hauv kev kawm tam sim no; Qhov tso tawm ntawm CPhGs tom qab 12 teev tau zoo li qub rau 100 feem pua, thaum lub CPhG liposomes ze rau 100 feem pua tom qab 24 teev, thiab qhov tso tawm ntawm CPhGs nce nrog lub sijhawm. Cov ntaub ntawv tau txheeb xyuas los ntawm kev siv cov qauv sib txawv rau cov kev tswj hwm kev tso tawm, suav nrog kev txiav txim xoom, xaj thawj zaug, thiab Higuchi sib npaug. Qhov sib npaug sib npaug thiab cov coefficients sib raug zoo rau peb qhov kev tso tawm qauv yog qhia hauv Table 1.
Cov qauv zoo tshaj plaws haum rau CPhGs thiab CPhG liposomes yog Higuchi thiab thawj qhov sib npaug. Kev tshawb fawb hauv vitro tso tawm ntawm CPhG liposomes tsis pom muaj qhov tshwm sim tawg. Qhov zoo tshaj plaws-fitting tis muaj nuj nqi raug xaiv los xam cov dissolution tsis T50 (kev puas tsuaj 50 feem pua). Cov txiaj ntsig tau pom tias lub sijhawm khaws cia nruab nrab ntawm CPhG liposomes ntev dua li ntawm CPhGs, thiab T50 ntawm CPhG liposomes yog 9.39 h. Piv nrog rau 1.69 h rau CPhGs, CPhG liposomes nthuav tawm cov txiaj ntsig zoo-tso tawm (Table 2).
Peb thawj zaug tshawb xyuas cov teebmeem cytotoxic ntawm CPhG liposomes ntawm HSCs nrog kev kuaj MTT hauv vitro. HSCs raug nthuav tawm rau ntau qhov sib txawv ntawm CPhG liposomes, xws li ntawm 0 txog 117.79 卩g/mL rau 24 teev. CPhG liposomes txo qhov kev muaj peev xwm ntawm HSCs nyob rau hauv koob tshuaj, thiab lawv cov nqi IC50 yog 42.535 (piv txwv li / mL. Cov nyhuv ntawm CPhG liposomes (29.45, 14.72, thiab 7.36 ^ g / mL) ntawm cov hlwb ntawm 24, 48 thiab 72 h yog pom nyob rau hauv daim duab 3. Tom qab kev kho mob nrog CPhG liposomes, qhov kev loj hlob ntawm HSCs tau cuam tshuam loj heev ntawm lub sijhawm sib txawv (Daim duab 3).
Lactate dehydrogenase (LDH) tso tawm ntawm HSCs tau kuaj pom tom qab 24 teev ntawm kev kho mob nrog ntau qhov sib txawv ntawm CPhG liposome. Cov txiaj ntsig tau qhia hauv Table 3. HSCs kho nrog CPhG liposomes tsis muaj txiaj ntsig zoo rau LDH tso tawm.
2.4. CPhG Liposomes Induce HSCs Apoptosis thiab Arrest Cell Cycle
HSCs tau raug rau CPhG liposomes (29.45,14.72,7.36 卩g/mL) rau 24 h, thiab annexin-V-FITC / PE ob chav staining tau ua los kuaj apoptosis los ntawm ntws cytometry, raws li qhia hauv daim duab 4a. Tag nrho apoptosis ntawm cov hlwb tau nce ntxiv piv rau pawg tswj hwm (p < 0.05).="" tom="" qab="" 24="" teev="" ntawm="" kev="" kho="" mob="" ntawm="" hscs="" rau="" txhua="" pawg="" koob="" tshuaj,="" cphg="" liposomes="" (29.45="" ^g="" ml)="" tuaj="" yeem="" ua="" rau="" mob="" apoptosis="" lig="" ntawm="" hscs,="" thiab="" feem="" pua="" ntawm="" cov="" hlwb="" apoptotic="" lig="" tau="" siab="" dua="" li="" ntawm="" cov="" hlwb="" apoptotic="" thaum="" ntxov.="" qhov="" feem="" pua="" ntawm="" cov="" hlwb="" apoptotic="" thaum="" ntxov="" tau="" ntau="" dua="" li="" cov="" hlwb="" apoptotic="" lig="" hauv="" cphg="" liposome="" (14.72="" thiab="" 7.36="" |^g="" ml)-kho="" pab="" pawg.="" cov="" txiaj="" ntsig="" no="" qhia="" meej="" meej="" tias="" cphg="" liposomes="" ua="" rau="" muaj="" hepatocyte="" apoptosis,="" tshwj="" xeeb="" tshaj="" yog="" nyob="" rau="" theem="" kawg="" ntawm="" hscs.="" tsis="" tas="" li="" ntawd,="" tag="" nrho="" apoptosis="" tau="" nce="" nrog="" qhov="" ntau="" npaum="" ntawm="" cphg="" liposomes.="" cov="" txiaj="" ntsig="" no="" qhia="" tias="" cphg="" liposomes="" tuaj="" yeem="" cuam="" tshuam="" kev="" ua="" haujlwm="" ntawm="" hscs="" thiab="" mob="" siab="" fibrosis="" los="" ntawm="" inducing="" apoptosis="" ntawm="">
Txhawm rau txiav txim siab qhov cuam tshuam ntawm CPhG liposomes ntawm lub voj voog ntawm tes ntawm HSCs, HSCs tau kho nrog CPhG liposomes ntawm qhov ntau ntawm 29.45, 14.72, thiab 7.36 昭 / mL rau 24 h, thiab tshuaj xyuas los ntawm kev ntws cytometry (Daim duab 4b). Cov txiaj ntsig tau pom tias qhov feem pua ntawm cov hlwb hauv G0/G1 theem tau nce ntxiv (p < {{10}}.05),="" thaum="" feem="" pua="" ntawm="" cov="" hlwb="" hauv="" theem="" s="" yog="" qhov="" tseem="" ceeb.="" txo="" (p=""><0.05). ntxiv="" mus,="" qhov="" kev="" faib="" ua="" feem="" ntawm="" cov="" hlwb="" hauv="" g2="" m="" theem="" tau="" txo="" qis="" thaum="" piv="" rau="" pawg="" tswj="" hwm=""><0.05). taken="" together,="" these="" results="" exhibit="" the="" cell="" cycle="" modulatory="" activity="" of="" the="" cphg="" liposomes="" in="" hscs,="" which="" may="" relate="" to="" their="" anti-proliferative="" and="" apoptosis-inducing="">
2.5. St udy ntawm Mechanism (s) ntawm Kev Ua ntawm C PhG Liposomes Hauv Vitro
2.5.1. CPhG Liposomes Inhibit HSCs Proliferation los ntawm rrPDGF-BB Stimulation
Txhawm rau tshawb xyuas ntxiv txog qhov cuam tshuam ntawm CPhG liposomes ntawm kev loj hlob txhawb nqa los ntawm rrPDGF-BB ntawm HSCs, MTT kev soj ntsuam tau ua los ntsuas qhov muaj peev xwm ntawm HSCs uas tau kho nrog preset concentrations ntawm CPhG liposomes rau 24, 48, thiab 72 h ( Daim duab 5).
Tom qab 24 teev ntawm kev kho mob nrog HSCs txhawb nqa los ntawm rrPDGF-BB thiab 29.45,14.721 thiab 7.36 昭/mL ntawm CPhG liposomes, qhov ciaj sia taus ntawm cov hlwb hauv txhua pawg koob tshuaj yog 67.5 feem pua, 75.3 feem pua, thiab 89.2 feem pua, feem. Tom qab 48 teev, cov ciaj sia taus yog 49.2 feem pua, 58.6 feem pua, thiab 73.5 feem pua, raws li, nyob rau hauv lub hlwb
ntawm txhua pawg koob tshuaj. Tom qab 72 teev ntawm kev kho mob, cov ciaj sia taus ntawm cov hlwb hauv txhua pawg koob tshuaj yog 45.3 feem pua, 59.2 feem pua, thiab 79.2 feem pua. Cov txiaj ntsig tau pom tias CPhG liposomes muaj qhov cuam tshuam tseem ceeb ntawm HSCs txhawb nqa los ntawm rrPDGF-BB, thiab cov nyhuv inhibitory no tau pom tseeb dua li qhov koob tshuaj tau nce. Nws tau pom tias cov nyhuv inhibitory ntawm CPhG liposomes ntawm HSCs pab txhawb rau CPhGliposomo los tiv thaiv kev loj hlob.
2.5.2. Lub CPhG Liposomes inhibit HSC Ua Haujlwm Hauv Vitro
HSCs nyob rau hauv qhov chaw ntawm Disse tsim ECM cov khoom xws li collagen-1, (s-SMA, thiab collagen III. Lwm yam uas tswj kev txhim kho ECMs, xws li MMPs thiab TIMPs, kuj yog tsim los ntawm HSCs [5]. Txhawm rau tshawb xyuas ntxiv cov txheej txheem oi rrPDGF-BB-induced hauv HSCs, peb tau soj ntsuam cov kev qhia ntawm ECM-txog cov proteins, suav nrog collagen-1, Cosma, collagen III, MMP-1, thiab TIMP{{8} }. Daim duab 6a qhia tau tias CPhG liposomes (29.45,14.72, 7.36 4g/mL) pab pawg tuaj yeem txo qis mRNA qhia qib ntawm collagen-1, a-SMA, thiab TIMP{{18} } thiab nce qib mRNA ntawm MMP-1 ntau dua li rrPDGF-BB pab pawg.
Daim duab 6. Kev cuam tshuam ntawm CPhG liposomes ntawm kev nthuav qhia ntawm collagen-1, a-SMA, TIMP-1 , thiab MMP-1 hauv HSCs. (a ) mRNA qhia ntawm collagen-1, a-SMA, TIMP-1 , thiab MMP-1 hauv HSCs (RT-PCR assay). (b) Cov protein qhia ntawm collagen{{10}} thiab a-SMA. A, pawg tswj; B, pawg kho mob rrPDGF-BB; C, rrPDGF-BB ntxiv rau CPhG liposome 29.45 g / mL kho pab pawg; D, rrPDGF-BB ntxiv rau CPhG liposome 14.72(_ig/mL kho pab pawg; E, rrPDGF-BB ntxiv rau CPhG liposome 7.3 6 卩g/m L kho d pab pawg; Cov ntaub ntawv raug nthuav tawm raws li qhov txhais tau tias 土 SD . ** p < 0.01,="" sib="" txawv="" heev="" piv="" rau="" rrpdgf-bb-activated="" hscs="" pawg.="" #="" p="">< 0.05,="" sib="" txawv="" coirtpared="" rau="" pawg="" tswj.="" 6-actin="" tau="" siv="" los="" ua="" kev="" tswj="" hwm="" sab="">
CPhG liposomes kuj nce qib mRNA ntawm MMP-1. Cov theem ntawm a-SMA thiab collagen-1 tau qhia ntau heev hauv rrPDGF-BB-activated HSCs. Hauv qhov sib piv, CPhG liposomes txo cov qib ntawm a-SMA thiab collagen-1 (Figure6b). Tshwj xeeb tshaj yog nyob rau hauv ib tug siab concentration (29.45 ^g / mL), CPhG liposome txo qhov nce collagen{10}} thiab a-SMA qhia ntawm rrPDGF-BB.
2.53 CPhG Liposomes Inhibit the PI3K/Akt Signaling Pathway
PI3K / Akt yog ib qho tseem ceeb ntawm cov teeb liab hloov txoj kev kho los ntawm tyrosine kinase receptors. PI3K / Akt signaling txoj kev ua raws li lub sensor nyob rau hauv cov lus teb rau extracellular stimuli thiab kho cov cellular signals, yog li ua lub luag hauj lwm tseem ceeb hauv cell apoptosis thiab proliferation los ntawm kev cuam tshuam cov kev ua ntawm downstream effector molecules [13]. Txhawm rau txheeb xyuas seb txoj hauv kev PI3K / Akt puas koom nrog kev tiv thaiv kev loj hlob ntawm CPhG liposomes ntawm HSCs, cov lus qhia thiab qib phosphorylation ntawm PI3K / Akt tau tshuaj xyuas los ntawm kev txheeb xyuas sab hnub poob. Cov txiaj ntsig tau pom tias cov qib ntawm p-PI3K thiab p-Akt tsis tu ncua hauv HSCs tom qab 24 teev kho nrog CPhG liposomes, qhia tau hais tias kev tiv thaiv kev loj hlob ntawm CPhG liposomes tiv thaiv HSCs cuam tshuam rau kev tsis ua haujlwm ntawm PI3K / Akt txoj hauv kev. , raws li qhia hauv daim duab 7.
2.5.4 ib. Lub CPhG Liiposomes tuaj yeem txo qis-Tshaj tawm ntawm PI3K / Akt Pathway Proteins hauv HSCs ntawm rrPDGF-BB-Mediated FAK Overexpression Plasmid
FAK yog cov noob tseem ceeb uas tswj cov PI3K/Akt teeb liab txoj hauv kev. Activated FAK tuaj yeem phosphorylate PI3K, uas txuas ntxiv mus rau phosphorylation ntawm Akt. FAK kho cov teeb liab hloov pauv ntawm tyrosine-protein kinase receptor, integrin, thiab lwm txoj hauv kev rau cov hlwb, uas yog qhov sib txuas thiab hub ntawm ntau txoj hauv kev hauv lub cev thaum HSCs ua kom. Yog li ntawd, peb speculated
tias CPhG liposomes tuaj yeem cuam tshuam kev ua haujlwm ntawm PI3K / Akt teeb liab txoj hauv kev los ntawm kev cuam tshuam qhov ua kom FAK. Tom qab ntawd peb tau lees paub qhov kev xav no los ntawm kev ua kom pom qhov overexpression ntawm FAK noob.
(pEX{{0}}NC) ntxiv rau CPhG liposome 29.45 Lg/mL pawg poob qis (p <0.01).
estern blotting nrog tshwj xeeb thawj cov tshuaj tiv thaiv. ** p < 0.01,="" sib="" txawv="" heev="" piv="">
rrPDGF-BB stimulation plus pEX-3-FAK pawg. ntxiv rau p < 0.05,="" sib="" txawv="" heev="" piv="" nrog="" rrpdgf-bb="" pab="" pawg="" kho.="" &="" p=""><0.01, sib="" txawv="" heev="" piv="" nrog="" rrpdgf-bb="">
plus pEX-3-NC pab. -actin tau siv los ua kev tswj hwm sab hauv.
cistanche tuaj yeem tiv thaiv mob qog noj ntshav
3. Kev sib tham
CPhGs tau pom tias muaj ntau yam kev ua haujlwm lom neeg, suav nrog kev ua haujlwm tiv thaiv kab mob [14,15], kev tiv thaiv kab mob osteoporosis [16,17], sedative nyhuv [18], kev ua kom qaug zog [19], cov nyhuv neuroprotective [ 20], thiab kev ua haujlwm hepatoprotective [21,22]. Yang et al. pom tias ib qho extract ntawmC. tubulosatuaj yeem ncua qhov kev nce qib ntawm kev sim tshuaj daim siab fibrosis los ntawm inhibiting collagen synthesis thiab txo oxidative kev nyuaj siab [22]. Txawm li cas los xij, qhov ua tau zoo ntawm CPhGs li
Cov tiv thaiv cua sov yog txwv los ntawm lawv cov permeability tsis zoo thiab tsis muaj bioavailability [23]. Yog li ntawd, nws yog ib qho tsim nyog los nthuav dav daim ntawv thov ntawm CPhGs los ntawm kev tsim kom muaj kev xa khoom zoo los kov yeej cov teeb meem no.
Nyob rau hauv xyoo tas los no, ob qho tib si kev tshawb fawb tshuaj thiab kev tshawb fawb soj ntsuam tau lees paub tias kev siv nanoparticles raws li cov neeg nqa tshuaj tuaj yeem txhim kho kev siv tshuaj hauv vivo. Liposomes yog qee qhov tseem ceeb tshaj plaws nanoparticles [24].
Liposomes tuaj yeem siv los ua cov neeg nqa tshuaj kom ua tiav cov tshuaj tiv thaiv kab mob, tshuaj tiv thaiv kab mob, tshuaj tua kab mob, tshuaj tua kab mob, thiab lwm yam tshuaj sib txawv [25]. Liposomes kuj tuaj yeem txhim kho kev ruaj ntseg tshuaj, nce tshuaj solubility, thiab txhawb biocompatibility [26]. Ntawm qhov tod tes, cov kev tshawb fawb tau pom tias cov tshuaj xa khoom ntawm cov khoom nanoparticles (nyob rau hauv thaj tsam ntawm 10-1000} nm) tuaj yeem mloog tau 80 feem pua ntawm cov tshuaj ntau npaum li cas hauv daim siab, ua kom cov tshuaj nkag mus. lub cev ntawm lub cev [27] Jing Zhu et al. pom tias galanin-liposomes muaj lub siab lub hom phiaj zoo [28]. Yog li ntawd, cov txiaj ntsig tsis zoo ntawm cov tshuaj nano-Suav tuaj yeem siv los kho cov kab mob siab lossis lwm yam kab mob. Qhov loj me me ntawm CPhG liposomes npaj rau hauv txoj kev tshawb no, uas muaj qee qib ntawm daim siab lub hom phiaj, yog (216.7 ± 3.47) nm. Tus cwj pwm tso tshuaj yeeb tshuaj yog ib qho tseem ceeb ntawm cov tshuab xa tshuaj nano. Vim tias CPhGs muaj cov dej solubility zoo, PBS tsis haum tshuaj (pH=60) yog siv los ua qhov nruab nrab tso tawm. Los ntawm kev tso tawm nkhaus, nws tuaj yeem pom tau tias ua ntej 2 h, qhov feem pua ntawm kev tso tawm ntawm CPhG liposomes tsawg dua 40 feem pua, tsis muaj kev tso tawm tam sim ntawd, thiab muaj qee yam kev txhawb nqa-tso tawm piv nrog kev tso tawm ntawm CPhGs. Los ntawm qhov sib npaug tso tawm nkhaus, kev tso tawm ntawm CPhG liposomes hauv cov tshuaj phosphate buffer ntawm pH=6.0 yog ua raws li Higuchi equation.
cistancheua tauanti-apoptosis
Hauv cov kab mob siab ntev, kev ua kom tsis tu ncua ntawm HSCs ua rau daim siab fibrosis [29]. Yog li ntawd, inhibiting qhov ua kom, proliferation, thiab cell voj voog ntawm HSCs, los yog inducing HSC apoptosis, tej zaum yuav yog ib tug tshiab pib taw tes rau lub hom phiaj kho mob ntawm daim siab fibrosis [30-33]. Ntau qhov kev tshawb fawb tau pom tias apoptosis yog tus yuam sij rau thim rov qab cov kab mob siab los ntawm kev txhawb nqa apoptosis [34]. Txoj kev loj hlob ntawm daim siab fibrosis nyob ntawm qhov ua kom HSCs, uas yuav muaj peev xwm inhibit HSC ua kom thiab induce apoptosis los tiv thaiv lossis kho daim siab fibrosis [35,36]. LDH yog ib qho enzyme nyob rau hauv lub cell cytosol, uas yog tso tawm nyob rau hauv kab lis kev cai xov xwm thaum lub cell membrane puas. Rau cov laj thawj no, peb tau ua hauv vitro kev tshawb fawb ntawm cov cell viability rates, cov ntsiab lus ntawm LDH hauv hlwb, thiab apoptosis thiab cell voj voog ntawm HSCs raug rau CPhG liposomes. Cov txiaj ntsig tau pom tias CPhG liposomes muaj zog tiv thaiv kev muaj peev xwm thiab kev loj hlob ntawm HSCs thiab nce feem pua ntawm cov hlwb apoptotic nyob rau hauv qhov concentration-dependent. Ntxiv mus, cov txiaj ntsig ntawm lub voj voog ntawm tes qhia tau tias CPhG liposomes tuaj yeem ua rau lub voj voog ntawm tes raug ntes nyob rau theem G1. Cov txiaj ntsig tau pom tias tus yuam sij rau CPhG liposomes los tiv thaiv thiab kho kab mob siab yog los tswj cov cell proliferation, apoptosis, thiab cell voj voog. Piv nrog rau pawg tswj hwm dawb paug, LDH cov ntsiab lus ntawm txhua pab pawg muaj qhov sib ntxiv zuj zus. Txawm li cas los xij, tsis muaj qhov sib txawv tseem ceeb (p > 0.05), qhia tau hais tias txawm hais tias qhov kev ua ub no txo qis rau HSCs cuam tshuam los ntawm CPhG liposomes, feem ntau ntawm cov cell membranes tsis zoo. Qhov inhibition ntawm kev loj hlob ntawm HSCs los ntawm CPhG liposomes tsis yog vim muaj tshuaj lom ntawm cov tshuaj rau cov hlwb.
PDGF yog qhov tseem ceeb tshaj plaws uas cuam tshuam rau kev loj hlob ntawm HSCs. Cov PDGF-receptors (PDGF-Ra thiab -p) belongs rau tsev neeg tyrosine kinase receptors. PDGF-AA khi tshwj xeeb rau PDGF-Ra, thaum PDGF-B chains khi thiab dimerize nrog PDGF-Ra thiab -p [37]. Tom qab khi ntawm ligand rau PDGF-R, cov receptors dimerize, uas tom qab ntawd ua rau phosphorylation ntawm cov tyrosine residue sab hauv thiab ua kom muaj ntau txoj hauv kev taw qhia, thaum kawg ua rau kev loj hlob thiab kev tsiv teb tsaws ntawm HSCs qhib. Cov txheej txheem no ua rau muaj kev tsim tawm ntau dhau thiab tso tawm ntawm collagen thiab lwm yam ECMs, yog li txhawb kev loj hlob thiab kev loj hlob ntawm daim siab fibrosis [38]. Breitkopf et al. qhia tau hais tias lub siab overexpression ntawm PDGF-BB ua rau HSC proliferation thiab daim siab fibrosis [39]. Hauv peb txoj kev tshawb fawb, peb tau siv rrPDGF-BB ua ib qho stimulator los ua kom HSCs, thiab txhua pawg CPhG liposome tuaj yeem cuam tshuam qhov kev loj hlob ntawm HSCs thiab nthuav tawm qhov pom tseeb ntawm kev sib raug zoo ntawm koob tshuaj. Nws tau pom tias cov nyhuv inhibitory ntawm CPhG liposomes ntawm HSCs pab txhawb rau CPhG liposomes 'kev tiv thaiv kev loj hlob.
HSCs ua kom muaj feem cuam tshuam nrog ntau yam kev hloov pauv hauv cov qauv qhia cov noob ntawm cov hlwb. Tshwj xeeb, kev hloov pauv hauv cov noob qhia ntawm collagen-1 thiab a-SMA feem ntau cuam tshuam nrog cov khoom fibrosis ntawm activated HSCs [40]. MMPs yog zinc-dependent endopeptidases uas ua lub luag haujlwm tseem ceeb hauv kev degradation ntawm tag nrho cov ECM cov protein nyob rau hauv lub cev thiab pathological mob. Cov MMPs thiab TIMPs kho kom haum rau kev sib xyaw thiab degradation ntawm ECM, feem. MMPs tuaj yeem txhawb ECM degradation. Qhov sib npaug ntawm MMPs thiab TIMPs txiav txim siab qhov tshwm sim ntawm daim siab fibrosis [29]. Piv nrog rau pawg rrPDGF-BB, kev qhia ntawm TIMP-1, a-SMA, thiab collagen-1 hauv pawg CPhG liposome tau nce ntau, thaum qhov kev qhia ntawm MMP-9 poob qis. CPhG liposome kev kho mob tshwj xeeb tau thim rov qab cov kab lus txawv txav uas tshwm sim los ntawm rrPDGF-BB hauv HSCs. Cov ntaub ntawv no qhia tau hais tias CPhG liposomes tuaj yeem cuam tshuam cov tiam ntawm ECM los ntawm kev rov qab qhov sib npaug ntawm MMPs thiab TIMPs, yog li nce qib ntawm MMP-1 thiab txo qhov kev qhia ntawm TIMP-1, a-SMA, thiab collagen{14}} thiab muaj peev xwm txhawb nqa cov cellular matrix degradation.
FAK yog hom focal adhesion complex. PDGF tseem tuaj yeem qhib FAK los ntawm kev cuam tshuam nrog ECM protein los ntawm integrin [41]. Lub focal adhesion complex muab ib tug ncaj qha sensor rau kev ncaj ncees ntawm lub extracellular ib puag ncig. Kev ua kom FAK ua rau kev ua kom PI3K. PI3K tau pom tias muaj kev koom tes hauv kev loj hlob ntawm ntau hom cell [42]. Nws lub luag haujlwm tseem ceeb hauv HSCs tau pom los ntawm kev siv PI3K-tshwj xeeb inhibitors LY294002 thiab wortmannin los thaiv PDGF-induced mitogenesis thiab chemotaxis yam tsis muaj kev cuam tshuam rau PDGF receptor autophosphorylation [43]. PI3K yog recruited rau hauv dimerized phosphorylated PDGF receptors, uas ua kom cov protein kinase C (PKC), Akt, thiab p70S6 kinases (p70S6K) [44]. Nws tau raug tshaj tawm tias qhov tseem ceeb-tsis zoo FAK (Ad-FAKCD) tau siv los thaiv cov haujlwm FAK thiab inhibit PI3K ua kom raug ntxias los ntawm PDGF thiab HSCs kev loj hlob tom qab PDGF kho [45]. Yog li, peb xav tias txoj hauv kev FAK / PI3K / Akt yog lub hom phiaj ua tau rau CPhG liposomes.
Peb cov txiaj ntsig tau pom tias piv rau rrPDGF-BB stimulation ntxiv rau X-3-FAK pawg, cov protein qhia theem ntawm FAK, phosphorylated PI3K, thiab phosphorylated Akt tau downregulated hauv rrPDGF-BB stimulation ntxiv rau X-3-FAK ntxiv rau CPhG liposome 29.45 Lg / mL pawg. Cov txiaj ntsig no qhia tau tias CPhG liposome 29.45 Lg / mL kev kho mob tuaj yeem txo qhov kev qhia ntawm FAK protein thiab phosphorylated PI3K thiab Akt proteins downstream ntawm FAK.
Nrog rau qhov tob ntawm kev tshawb fawb ntawm chemistry thiab pharmacology ntawm cov tshuaj suav tshuaj, ntau thiab ntau cov khoom xyaw ntawm cov tshuaj suav tshuaj tau pom thiab paub tseeb. Paclitaxel, artemisinin, thiab lwm yam tshuaj tau dav lees paub tias yog thawj kab tshuaj kho mob rau cov kab mob cuam tshuam thoob ntiaj teb. Txawm li cas los xij, tib lub sijhawm, nws tau pom tias txawm hais tias cov tshuaj hauv vitro pharmacological ntawm ntau cov khoom siv kho mob hauv Suav teb muaj zog heev, lawv muaj cov dej solubility tsis zoo, lub neej luv luv, tsis muaj kev ruaj ntseg, tsis muaj bioavailability, tshuaj lom, thiab ntau yam. lwm yam teeb meem uas txwv tsis pub lawv cov tshuaj thiab tshuaj kho mob. Lub nano-drug xa system yog ib qho kev xa tshuaj tshiab nrog lub peev xwm loj rau kev txhim kho thiab yog qhov hotspot hauv kev tshawb fawb tshuaj niaj hnub. Nws daim ntawv thov nyob rau hauv kev tshawb fawb thiab kev loj hlob ntawm cov ntaub ntawv tshiab ntawm tsoos suav tshuaj tsis tau tsuas yog txhim kho cov tsoos cov ntaub ntawv ntawm Suav tshuaj tab sis kuj txhim kho cov curative nyhuv ntawm tsoos suav tshuaj rau ib tug tej yam thiab ua rau kom cov stability ntawm cov tshuaj nyob rau hauv vivo. Ntxiv mus, cov liposomes uas tsis tau hloov kho feem ntau yog absorbed los ntawm RES, nyob ntawm seb lawv qhov loj me, cov khoom nto, thiab lwm yam. Lawv kuj yog passively tsom rau lub siab. Txhawm rau txhim kho lub siab lub hom phiaj kev ua tau zoo ntawm liposomes zoo tib yam, qhov concentration ntawm cov ntaub so ntswg tuaj yeem nce ntxiv los ntawm kev sib txuas cov ligands tshwj xeeb lossis cov pab pawg ua haujlwm tshwj xeeb.
Yog li ntawd, hauv kev tshawb fawb yav tom ntej, peb yuav tsum muab qhov tseem ceeb rau lub cev thiab tshuaj lom neeg ntawm CPhG extraction thiab purification, xws li solubility, stability, acidity, thiab alkalinity, nrog rau kev kawm txog biopharmaceutics thiab pharmacokinetics. Peb yuav tsum tsim kom muaj ib lub tswv yim tsim qauv tsim tshwj xeeb, kev npaj thev naus laus zis platform, thiab kev soj ntsuam zoo rau CPhGs thiab tseem tsim ntau yam nano-drug-loading systems rau CPhGs, kom ntseeg tau tias CPhGs tau siv kom zoo li sai tau. Kev siv tag nrho cov txiaj ntsig no yuav yog kev coj ua ntawm peb txoj kev tshawb fawb yav tom ntej.
