Manganese(II) Complexes Stimulate Antitumor Immunity Via Aggravating DNA puas thiab ua kom lub CGAS-STING txoj kev

Ua kom lub cyclic GMP-AMP synthase-stimulator ntawm interferon gene (cGAS-STING) txoj kev yog ib qho kev cog lus tiv thaiv kab mob rau kev kho mob qog noj ntshav. Manganese(II) complexes MnPC thiab MnPVA (P=1), 10-phenanthroline, C=chlorine, thiab VA=valproic acid) tau pom los qhib txoj hauv kev cGAS-STING . Cov complexes tsis tsuas yog puas DNA, tab sis kuj inhibit histone deacetylases (HDACs) thiab polyadenosine diphosphate-ribose polymerase (PARP) los cuam tshuam kev kho DNA puas, yog li txhawb nqa cov xau ntawm DNA tawg mus rau hauv cytoplasm. Cov DNA tawg tau qhib txoj hauv kev cGAS-STING, uas tau pib lub cev tiv thaiv kab mob hauv lub cev thiab kev sib txuas lus ob txoj hauv kev ntawm cov qog hlwb thiab cov kab mob sib kis. Lub tshuab ua kom cGAS-STING ntxiv rau kev tsim tawm ntawm hom I interferons thiab tso tawm ntawm pro-inflammatory cytokines (TNF-a thiab IL-6), boosting cov qog infiltration ntawm dendritic hlwb thiab macrophages, nrog rau stimulating cytotoxic T hlwb. tua cov qog nqaij hlav hauv vitro thiab hauv vivo. Raws li kev txhim kho DNA-kev muaj peev xwm ua kom puas tsuaj, MnPC thiab MnPVA tau pom muaj zog tiv thaiv kab mob thiab kev tiv thaiv kab mob ntau dua li Mn2+ ions, yog li qhia tau tias muaj peev xwm zoo li cov tshuaj chemoimmunotherapeutic rau kev kho mob qog noj ntshav.

Cov txiaj ntsig ntawm cistanche tubulosa-Antitumor

Taw qhia

Kev rov tshwm sim dua, metastasis, thiab tshuaj tiv thaiv ntawm cov qog ua rau muaj teeb meem loj rau cov pa tshuaj tiv thaiv kab mob.1 Immunotherapy yog ib lub tswv yim zoo los tshem tawm cov qog hlwb los ntawm kev siv los yog txhawb lub cev tiv thaiv kab mob ntawm cov neeg mob.2,3 Xyoo dhau los, ntau yam kev kho mob qog noj ntshav nrog rau Cov tshuaj tiv thaiv kab mob tiv thaiv kab mob, kab mob oncolytic, thiab chimeric antigen receptor T (CAR-T) cov hlwb tau siv rau hauv kev kho mob los txhim kho kev tiv thaiv kab mob tiv thaiv kab mob.4–6 Txawm li cas los xij, thawj zaug thiab hloov tshuaj tiv thaiv, kev tiv thaiv kab mob tsis txaus, thiab poob ntawm cov qog tshwj xeeb. Antigen lub hom phiaj feem ntau ua rau muaj kev cuam tshuam ntawm kev siv tshuaj tiv thaiv kab mob.7,8 Kev tiv thaiv kab mob tiv thaiv kab mob yog nyob ntawm kev tiv thaiv kab mob hauv lub cev.8 Dhau li ntawm molding thiab txhawb nqa kev tiv thaiv kab mob tiv thaiv kab mob, lub cev tsis muaj zog tiv thaiv kab mob hauv lub cev raws li qhov cuam tshuam rau kev tiv thaiv tus tswv tsev lub luag haujlwm tseem ceeb rau kev tiv thaiv kab mob. Kev lees paub ntawm cov qog hlwb.9 Hmoov tsis zoo, thaum lub sij hawm qog nqaij hlav, cov qog nqaij hlav malignant oen khiav tawm ntawm kev tiv thaiv kab mob thiab nws thiaj li loj hlob mus rau hauv cov qog "txias". cov lus teb rau cov qog thiab txhim kho cov nyhuv kho ntawm immunotherapy. Lub cyclic GMP-AMP synthase-stimulators ntawm interferon genes (cGAS-STING) tsim cov khoom tseem ceeb ntawm kev tiv thaiv kab mob hauv lub cev, uas tau tshwm sim los ua lub hom phiaj zoo rau kev tiv thaiv kab mob qog noj ntshav.12,13 Kev ua kom txoj hauv kev cGAS-STING ua rau muaj kev poob qis. Cov xwm txheej qhia, suav nrog kev txhawb nqa thiab nrhiav neeg ua haujlwm ntawm TANK-binding kinase 1 (TBK1) thiab interferon regulatory factor 3 (IRF3), uas ua rau muaj kev tso tawm thiab tso tawm ntawm hom I interferons (IFN-Is) thiab proinflammatory yam IL-6 thiab TNF- ua. 13,14 IFNs tom qab txhawb kev loj hlob thiab kev tsiv teb tsaws ntawm dendritic hlwb (DCs), txhim kho lub ntuj tua neeg (NK) cell-mediated cytotoxic nyhuv, thiab ntoo khaub lig-prime qog specic T hlwb, yog li orchestrating lub innate thiab adaptive tiv thaiv kom tswj tus cwj pwm. ntawm aggressive qog.15,16 Tam sim no, cov txheej txheem kho ntawm me me molecule qhov ncauj agonist MSA-2,17 natural agonist cyclic dinucleotides (CDNs), 18 thiab qee qhov nano-systems19–23 tau raug sim hauv murine qog qauv rau cuam ​​tshuam rau hauv txoj kev cGAS-STING. Manganese (Mn) yog ib qho khoom noj muaj txiaj ntsig uas ua lub luag haujlwm tseem ceeb hauv ntau lub cev lub cev nrog rau cov tshuaj tiv thaiv kab mob tiv thaiv kab mob.24,25 Tsis ntev los no, Mn2+ ions tau tshawb pom los txhim kho qhov rhiab heev ntawm ob-strand DNA (dsDNA) sensor. cGAS thiab ua rau kev tsim cov tub txib thib ob cGAMP, yog li ua kom STING kev ua si los ntawm augmenting cGAMP-STING binding affinity.12,26 Ntxiv mus, Mn2+ ions indirectly inhibit qog hlav los ntawm kev txhawb txoj haujlwm ntawm CD8+ T hlwb ntawm inducing IFN ntau lawm hauv vivo. 27 Txawm li cas los xij, kev tswj hwm ncaj qha ntawm Mn2+ ions hauv vivo tsis tuaj yeem lav qhov muaj txiaj ntsig zoo hauv cov qog microenvironment23 thiab ntau dhau Mn2+ ions tuaj yeem ua rau cov kab mob toxicity, xws li irreversible neurotoxicity thiab cardiovascular toxicity.28 Mn complexes muaj ntau dua. ruaj khov thiab inert rau biomolecules vim muaj kev tiv thaiv ntawm ligands thiab li no tuaj yeem txo qhov toxicity ntawm Mn2+ ions. Txawm li cas los xij, tsis muaj Mn complex tau siv los qhib txoj hauv kev cGAS-STING txog tam sim no. Kev hloov pauv ntawm Epigenetic reversibly hloov cov kab lus gene, uas tuaj yeem ua rau kev pib thiab kev loj hlob ntawm cov qog nqaij hlav thiab kev tiv thaiv kev tiv thaiv kab mob. Kev hloov pauv hloov pauv ntawm epigenetic hloov pauv tso cai rau cov hlwb tsis zoo rov qab mus rau lub xeev qub.29,30 Histone deacetylases (HDACs) kho cov protein acetylation, chromatin dynamics, protein hloov pauv, thiab cov lus teb DNA puas. HDAC inhibitors yog ib chav kawm ntawm cov muaj zog epigenetic modulators, nrog chromatin raws li lawv lub hom phiaj, ua raws li feem ntau los yog tag nrho cov qog hom.31 Qhov inhibition ntawm HDACs tsawg kawg yog ib feem pab rau histone acetylation, ua rau hloov pauv tsim thiab kho DNA ob-strand so ( DSB), 32,33 uas yuav pab txhawb kom tshem tawm cov tshuaj tiv thaiv hauv cov qog nqaij hlav cancer.34 Cov qauv chromatin so kom raug ntxias los ntawm HDAC inhibitors tuaj yeem txhawb cov kws khomob kom nkag mus rau DNA yooj yim dua, yog li ua rau DNA puas tsuaj, 35 uas muaj txiaj ntsig zoo rau kev ua haujlwm ntawm the cGAS STING pathway. Qee qhov HDAC inhibitors-xws li valproic acid (VA)- cuam tshuam rau chav kawm I thiab II HDACs (HDAC1/2), uas ua rau mob qog noj ntshav rau DNA-kev kho mob.36

cistanche tubulosa- txhim kho lub cev tiv thaiv kab mob

Polyadenosine diphosphate-ribose polymerases (PARPs) yog cov proteins tseem ceeb koom nrog hauv kev mob qog noj ntshav rau cov kws khomob. Cov enzymes no tseem ceeb heev rau kev kho DNA ib leeg-strand so (SSBs), 37 thiab koom nrog ntau cov txheej txheem ntawm tes, xws li apoptosis, kev tiv thaiv kab mob, kev hloov cov noob, thiab o.38-40 PARP inhibitors tiv thaiv kev kho ntawm DNA lesions, ua rau kom muaj txiaj ntsig ntawm cytosolic DNA, uas tau lees paub los ntawm cGAS los ua kom lub cGAS-STING txoj hauv kev.41,42 A series ntawm anticancer hlau complexes suav nrog cov platinum, ruthenium, thiab kub tau pom tias inhibit qhov kev ua PARP.43, 44 Ntawm no peb qhia txog cov tshuaj tiv thaiv kab mob thiab tshuaj tiv thaiv kab mob ntawm ob lub MnII complexes MnPC thiab MnPVA. Cov txheej txheem tshuaj thiab siv lead ua ntawm cov complexes yog pom nyob rau hauv daim duab 1. Nyob rau hauv cov complexes, 1, 10-phenanthroline (P) yog ib tug nontoxic DNA intercalator, thiab VA yog ib tug inhibitor ntawm HADCs.31 VA yuav txhawb lub acetylation ntawm DNA-conjugated histone proteins, txhim khu kev nkag tau ntawm DNA nyob rau hauv so chromatin, thiab reactivate dormant qog suppressor genes.33,35 Peb xav tias kev koom ua ke ntawm 1,10-phenanthroline thiab/los yog VA rau hauv MnPC lossis MnPVA yuav muaj txiaj ntsig. cov complexes nrog antiproliferative kev ua si los ntawm inducing DNA puas thiab stimulating antitumor tiv thaiv. Ib qho kev sim ua tau pom tias MnPC thiab MnPVA ua rau DNA puas tsuaj zoo, ua rau txoj hauv kev cGAS-STING hauv cov qog thiab lub cev tiv thaiv kab mob, thiab nce qhov tso tawm ntawm IFNs thiab pro-inflammatory cytokines. Tshwj xeeb, MnPVA inhibited cov haujlwm ntawm HDAC1/2 thiab PARP1, yog li ua kom txoj hauv kev cGAS-STING zoo dua MnPC. Tag nrho cov txiaj ntsig tau lees paub ob qho tib si hauv vitro thiab hauv vivo. Rau qhov zoo tshaj plaws ntawm peb txoj kev paub, MnPC thiab MnPVA yog cov rest multifunctional MnII complexes uas suppress qog hlwb feem ntau los ntawm activating antitumor tiv thaiv ntawm DNA puas-pib cGAS-STING txoj kev.

Fig. 1 Tshuaj thiab siv lead ua qauv ntawm MnPC thiab MnPVA. Hydrogen atoms raug tshem tawm kom pom tseeb.

Cov txiaj ntsig thiab kev sib tham

Tshuaj thiab lub cev muaj zog

MnPC thiab MnPVA tau npaj raws li cov ntaub ntawv txoj kev 45,46 thiab tag nrho cov yam ntxwv los ntawm kev tsom xam, IR spectroscopy (Fig. S1†), electron paramagnetic resonance (Fig. S2, † EPR), thiab X-ray crystallography. Cov qauv siv lead ua tsis muaj thiab xaiv qhov ntev thiab cov kaum sab xis tau nthuav tawm hauv Tables S1 thiab S2 (saib ESI†). Lub Mn atom nyob rau hauv cov complexes tau nthuav tawm ib tug distorted octahedral geometry nrog ib tug sib koom tes ntawm 6. MnPC crystallized nyob rau hauv lub monoclinic crystal system nrog rau qhov chaw pab pawg neeg P21 / c; MnII tsim plaub Mn– N daim ntawv cog lus nrog ob daim ntawv cog lus 1,{{10}}phenanthroline ligands thiab ob daim ntawv cog lus Mn–Cl. Mn-N daim ntawv cog lus ntev sib txawv ntawm 2.281 thiab 2.369 Å, uas yog, daim ntawv cog lus ntev ntawm Mn–N1, Mn–N2, Mn–N3, thiab Mn–N4 yog 2.369(4), 2.281(3), 2.341(3). ), thiab 2.287(3) Å, ntsig txog, thiab lub kaum sab xis ntawm N1–Mn–N2, N1–Mn–N4, thiab N2– Mn–N3 yog 71.15(11) degree, 97.86(11) degree , thiab 89.00(11) degree , feem. Cov qauv no txawv me ntsis ntawm qhov uas tau tshaj tawm los ntawm Abbas li al., 45 qhov twg MnPC crystallized nyob rau hauv triclinic crystal system nrog rau qhov chaw pab pawg P1. Yog li ntawd, qhov sib txuas ntev thiab cov ces kaum sib txawv hauv cov xwm txheej no. Zoo ib yam li cov qauv qhia, 46 MnPVA crystallized nyob rau hauv lub monoclinic crystal system nrog rau qhov chaw pab pawg neeg C2 / c, muaj ib tug N2O4 pub teeb. MnII koom tes nrog ob lub N atoms los ntawm 1,10-phenanthroline, thiab plaub O atoms los ntawm ib qho dej molecule thiab ob lub VA ligands, feem. Ib qho ntawm VA reacted li monodentate ligand, thaum lwm yam yog ib tug bidentate ligand. Daim ntawv cog lus ntev ntawm Mn-N1 thiab Mn-N2 yog 2.265(19) thiab 2.298(2) Å, ntsig txog, thiab ntawm Mn-O1, Mn-O2, Mn-O3, thiab Mn-O5 yog 2.2476(19), 2.260 (2), 2.0639 (18), thiab 2.1471 (17) Å, ntsig txog. Raws li ib tug muaj zog N, Nchelating ligand, 1, 10-phenanthroline stabilizes Mn complexes tiv thaiv demetallation. Kev ruaj ntseg ntawm MnPC thiab MnPVA hauv phosphate-buffered saline (PBS, nrog 0.5% v/v DMSO, pH 7.4, thiab 37 degree) thiab cell culture media (muaj 10% FBS) tau tshawb xyuas los ntawm UV pom spectroscopy (Fig. S3† ). Lub sij hawm-dependent UV spectra qhia tau hais tias cov complexes ruaj khov nyob rau hauv 72 teev.

Table 1 IC50 qhov tseem ceeb (mM) ntawm MnPC thiab MnPVA tiv thaiv cov kab sib txawv ntawm 72 h, nrog MnCl2, VA, CDDP, thiab P raws li cov lus qhia. Cov ntaub ntawv tau qhia tias txhais tau tias ± tus qauv sib txawv (SD, n=3)

Antiproliferative kev ua ub no

Kev ua haujlwm tiv thaiv kab mob ntawm cov tshuaj tiv thaiv tib neeg lub mis triple-negative cancer (MDA-MB-231), human pancreatic cancer (PANC-1), human hepatocellular cancer (HepG2), nas cancer mis (4T1) cell kab, thiab tib neeg lub raum tubular epithelial (HK-2) cell kab raug soj ntsuam los ntawm MTT kev soj ntsuam. MnCl2, VA, cisplatin (CDDP), thiab 1,10-phenanthroline (P) tau siv los ua cov khoom siv siv. Qhov ib nrab-qhov siab tshaj plaws inhibitory concentrations (IC50) ntawm 72 h tau teev nyob rau hauv Table 1. MnPC thiab MnPVA tau pom muaj zog tiv thaiv kab mob rau tag nrho cov kab mob qog noj ntshav, tshwj xeeb tshaj yog rau MDA-MB-231 thiab PANC-1 cov hlwb uas tsis hnov ​​​​tsw rau CDDP, qhia txog 8- thiab 11- lub sijhawm ua haujlwm siab dua, feem. Kev tiv thaiv kev ua haujlwm ntawm MnPC thiab MnPVA tiv thaiv HepG2 thiab 4T1 zoo ib yam li CDDP. Interestingly, ob qho tib si MnPC thiab MnPVA pom qhov txo qis toxicity ntawm HK-2 hlwb, nrog rau IC50 qhov tseem ceeb yog me ntsis siab dua li ntawm CDDP. Los ntawm qhov sib txawv, MnCl2, VA, thiab P yog yuav luag tsis muaj tshuaj lom nyob rau hauv cov kab ntawm tes, uas tau pom zoo nrog cov ntaub ntawv.13,36 Raws li cov txiaj ntsig no, cov kev tshawb fawb hauv qab no tau tsom mus rau kev ua ntawm MnPC thiab MnPVA ntawm MDA-MB. -231 hlwb.

Cellular uptake thiab DNA puas

Kev sib sau ntawm cov complexes hauv MDA-MB-231 hlwb nyob rau hauv cov nqe lus ntawm Mn yog ntsuas los ntawm ICP-MS co-incubation rau 24 teev. Raws li pom nyob rau hauv daim duab 2A, cov tsub zuj zuj ua raws li ib qho kev txiav txim ntawm MnPVA > MnPC > MnCl2, ua raws li lawv cov kev tiv thaiv kev loj hlob. Peb tau txiav txim siab ntxiv DNA-bound Mn hauv MDA-MB-231 hlwb los ntawm ICP-MS. Raws li pom hauv daim duab 2B, MnPC thiab MnPVA tau nthuav tawm ntau dua DNA binding muaj peev xwm tshaj MnCl2 tej zaum vim lawv cov cellular uptake ntau dua. Phosphorylated histone g-H2AX yog ib qho rhiab cim ntawm DNA ob-strand so (DSBs).47 Txhawm rau soj ntsuam DNA puas raug ntxias los ntawm Mn complexes, peb tau kuaj pom qhov qhia ntawm gH2AX los ntawm immunoblotting. Raws li qhia hauv daim duab 2C, MnPC thiab MnPVA tau nce qhov kev qhia ntawm g-H2AX hauv MDA-MB-231 hlwb ntawm 24 teev. Nws paub tias hlau complexes ntawm P yog cov zoo DNA intercalators; 48,49 txawm li cas los xij, P ib leeg tsis tshua ua rau DNA puas (Daim duab S4†). Kev nthuav qhia ntawm g-H2AX hauv cov qog nqaij hlav ntawm 4T1 cov qog-cov kabmob nas kuj tau tshawb xyuas los ntawm kev siv tshuaj tiv thaiv kom muaj kev kho mob nrog txhua qhov sib xyaw (1.3 mg Mn ib kg) ib zaug txhua 2 hnub rau 16 hnub. MnPC thiab MnPVA ua rau cov DNA puas hauv cov qog nqaij hlav, thaum MnCl2 thiab PBS NW nyuam qhuav cuam tshuam g-H2AX (Fig. S5†). dsDNA tso tawm los ntawm MDA-MB- 231 hlwb mus rau kab lis kev cai nruab nrab supernatant tau txiav txim siab los ntawm kev siv cov khoom siv ELISA aer kho nrog Mn complexes. Raws li tau nthuav tawm hauv daim duab 2D, cov ntsiab lus ntawm dsDNA hauv MnPC- lossis MnPVA-kho cell kab lis kev cai nruab nrab yog pom tau tias siab dua li lwm pab pawg. Cov txiaj ntsig tau qhia tias MnPC thiab MnPVA tuaj yeem txhawb nqa qhov tsis ruaj khov ntawm genomic DNA thiab qib ntawm cytosolic DNA. Qhov xwm txheej ntawm DNA puas yog  ua ntej kawm los ntawm kev ntsuas qhov  fluorescence hloov ntawm calf thymus DNA-ethidium bromide (EB) system. EB yog ib qho intercalator uas ua rau muaj kev nce ntxiv hauv fluorescence thaum khi rau DNA, thaum lub fluorescence txo qis thaum nws hloov chaw.50 MnPC thiab MnPVA me ntsis txo cov fluorescence siv (Fig. S6†), qhia tias cov complexes yuav intercalate. mus rau hauv DNA grooves los hloov lub khi EB vim lub hom phiaj ntawm P. Txawm li cas los xij, kev sib cuam tshuam tsis yog covalent ruaj khov. Qee qhov Mn complexes tuaj yeem tsim intracellular reactive oxygen hom (ROS) thiab induce oxidative stress, 51 thiab peb thiaj li kuaj tau ROS hauv MDA-MB-231 hlwb los ntawm confocal imaging siv 2′,7′ -dichloro-fluorescein diacetate ( DCFH-DA) raws li ROS  fluorescence sojntsuam ib qho kev raug rau Mn complex rau 18 teev. Raws li pom nyob rau hauv daim duab 3, MnPC thiab MnPVA intensified lub intracellular ROS  fluorescence piv rau cov tswj los yog MnCl2, tshwj xeeb tshaj yog MnPVA, tawm tswv yim hais tias lawv yuav ua rau oxidative kev puas tsuaj rau cellular DNA. Qhov kev puas tsuaj rau DNA tsis yog tsuas yog ua rau kev tua cov qog nqaij hlav cancer xwb tab sis kuj txhawb kev tiv thaiv kab mob los ntawm kev ua rau cGAS-STING txoj hauv kev.14,52

Fig. 2 Cellular uptake (A) thiab DNA-bound Mn (B) txiav txim los ntawm ICPMS, qhia txog DNA puas marker g-H2AX txiav txim los ntawm western blotting (C), thiab extracellular dsDNA txiav txim los ntawm kev siv cov khoom siv ELISA (D) tom qab MDA-MB-231 hlwb raug kho nrog MnCl2, MnPC, MnPVA, thiab VA (6 mM) raws li 24 teev.

Cell voj voog ntes thiab apoptosis

Cellular DNA puas teb (DDR) yog txuam nrog cov teeb liab uas tsav lub checkpoint capture ntawm lub voj voog ntawm tes.53 Qhov cuam tshuam ntawm Mn complexes ntawm lub voj voog ntawm tes ntawm MDA-MB-231 hlwb tau soj ntsuam los ntawm ow cytometry. Raws li pom nyob rau hauv daim duab 4A, MnPC, thiab MnPVA me ntsis nce lub G1 theem raug ntes thaum ua rau tag nrho lub cev qhuav dej ntawm G2 theem piv rau kev tswj thiab MnCl2. Cov txiaj ntsig tau qhia tias DNA kev puas tsuaj tshwm sim los ntawm MnPC thiab MnPVA cuam tshuam rau theem tom qab ntawm DNA synthesis. Li no, cov txheej txheem tiv thaiv kab mob ntawm MnPC thiab MnPVA txawv ntawm MnCl2. Hom kev tuag ntawm MDA-MB-231 hlwb tau tshawb xyuas los ntawm ow cytometry aer kho nrog cov complexes rau 72 h thiab staining nrog annexin V-FITC thiab propidium iodide (PI). Raws li qhia hauv daim duab 4B thiab S7,† hauv kev sib piv nrog kev tswj hwm thiab MnCl2, apoptosis (thaum ntxov + lig) tus nqi raug ntxias los ntawm MnPC thiab MnPVA mus txog 84.0% thiab 87.4% feem. Cov txiaj ntsig tau qhia tias MnPC thiab MnPVA muaj lub peev xwm pro-apoptotic muaj zog, uas cuam tshuam nrog lawv cov kev ua haujlwm antitumor.

cistanche ntxiv cov txiaj ntsig-nce kev tiv thaiv

Mitochondrial membrane muaj peev xwm

Mitochondria yog cov neeg koom nrog hauv lub cev tiv thaiv kab mob, thiab mitochondrial DNA (mtDNA) tau lees paub tias yog ib qho agonist ntawm lub cev tiv thaiv kab mob.54 Nws tau tshaj tawm tias ob qho tib si cytosolic DNA thiab mtDNA khi rau cov qauv-paub cov receptors thiab ua rau cGAS-STING teeb liab txoj hauv kev. 55 Txhawm rau tshawb nrhiav qhov muaj peev xwm mtDNA tso tawm los ntawm mitochondria nyob rau hauv lub xub ntiag ntawm MnPC thiab MnPVA, peb tau tshawb xyuas qhov ncaj ncees ntawm cov mitochondrial membrane los ntawm confocal imaging siv JC-1 raws li ib tug  fluorescent sojntsuam, uas ua aggregates thiab qhia liab fluorescence nyob rau hauv ib txwm mitochondria nrog lub siab DJm, thaum tshwm sim raws li ib tug monomer thiab muab tawm ntsuab  fluorescence nyob rau hauv puas cov uas tsis muaj DJm. 56 Raws li pom nyob rau hauv daim duab 5, MnPC thiab MnPVA txo cov kev siv ntawm cov liab fluorescence nyob rau hauv MDA-MB-231 hlwb stained nrog JC-1. Qhov "ntsuab (G) rau liab (R)" fluorescence piv rau MnPC- thiab MnPVA-kho cov hlwb yog 4.91 thiab 5.66, raws li, ntau dua li qhov kev tswj hwm (2.35) thiab MnCl2- kho hlwb ( 1.75 Nws.). Cov kev soj ntsuam pom tau hais tias kev ncaj ncees ntawm mitochondrial membrane raug puas tsuaj los ntawm MnPC thiab MnPVA, uas tuaj yeem pab txhawb kev xau ntawm mtDNA rau hauv cytosol kom ua rau txoj hauv kev cCAS-STING.

Fig. 3 Fluorescence confocal imaging ntawm ROS generated los ntawm Mn complexes (12 mM) hauv MDA-MB-231 hlwb tom qab incubation rau 18 h thiab staining nrog DCFH-DA (10 mM) rau 30 min.

Fig. 4 Cell voj voog ntes ntawm MDA-MB-231 hlwb tom qab incubation nrog sib txawv compounds (6 mM) rau 24 h (A), thiab apoptotic tsom xam ntawm MDA-MB-231 hlwb los ntawm ntws cytometry tom qab incubation nrog sib txawv tebchaw (6 mM) rau 72 h (B).

Kev ua thiab kev qhia ntawm HDACs

HDAC inhibitors sensitize cov qog nqaij hlav cancer rau DNA-kev kho mob los ntawm kev hloov cov qauv chromatin thiab cuam tshuam DNA kho.33,35 Raws li chav kawm I HDAC inhibitor, VA xaiv lub hom phiaj ntawm HDAC1 thiab HDAC2 enzymes, modulating DNA kev puas tsuaj taw qhia los rhuav tshem genomic stability thiab inhibit qog. kev loj hlob hauv vivo. 31 Kev koom ua ke ntawm VA hauv MnPVA tuaj yeem txhawb nqa qhov inhibition ntawm HDAC thiab induction ntawm DDR zoo dua. Yog li, peb txiav txim siab qhov cuam tshuam ntawm Mn complexes ntawm tag nrho HDAC kev ua hauv MDA-MB-231 hlwb tom qab coincubation rau 48 teev. Raws li pom nyob rau hauv daim duab 6A, MnPVA ho inhibited tag nrho HDAC kev ua si, nrog rau inhibitory kev ua si yog li ntawm 1.5 npaug ntau dua li ntawm VA. Peb tau kawm ntxiv txog qhov qhia txog HDAC1/2 cov proteins hauv MDA-MB-231 hlwb los ntawm immunoblotting. Raws li pom nyob rau hauv daim duab 6B thiab C, MnPVA obviously downregulated cov kev qhia ntawm HDAC1 thiab HDAC2 raws li piv nrog rau kev tswj. Txawm hais tias MnPC inhibited HDAC kev ua ub no, nws tsis tshua muaj feem cuam tshuam rau kev qhia ntawm HDAC1/2 proteins. Tsis xav txog, VA raws li tus paub HDAC inhibitor tsis qhia pom tseeb inhibition ntawm HADC1/2 ntawm qhov concentration (6 mM), yog li qhia txog lub luag haujlwm tseem ceeb ntawm Mn kev sib koom tes hauv kev ua kom muaj zog inhibition ntawm VA ntawm HADC. Cov txiaj ntsig tau qhia tias MnPVA yog ib qho zoo inhibitor ntawm HDAC1/2, uas yuav txhawb nqa DDR thiab ua rau txoj hauv kev cGAS-STING, yog li txhawb kev tiv thaiv kab mob. Peb tau tshawb pom ntxiv txog qhov qhia txog HDAC hauv cov qog nqaij hlav ntawm 4T1 cov qog-cov kabmob nas los ntawm kev tiv thaiv kab mob kev tiv thaiv kev kho mob nrog txhua qhov sib xyaw. Raws li pom hauv daim duab 6D, MnPVA pom tseeb txo qis ntawm HDAC1, thaum MnPC thiab MnCl2 tsuas yog me ntsis lossis cuam tshuam HDAC1 xwb.

Cov txiaj ntsig ntawm cistanche tubulosa-Antitumor

Kev nthuav qhia ntawm PARP1 thiab apoptotic proteins

PARPs ua lub luag haujlwm tseem ceeb hauv kev kho cov kab mob DNA. PARP inhibitors tiv thaiv DNA kho, uas ua rau kev xa tawm ntawm DNA tawg mus rau cytosol, ua rau cGAS-mediated innate immune teb.42 Raws li cov protein sensor rau DNA strand so, PARP1 localizes mus rau qhov chaw ntawm DNA puas thiab dominates kho ntawm DNA, yog li dhau los ua lub hom phiaj tseem ceeb rau kev kho mob qog noj ntshav. Kev ua kom cov caspases, tshwj xeeb tshaj yog caspase-3, tuaj yeem pib apoptosis los ntawm cleaving PARP1, uas yog suav tias yog ib qho cim ntawm apoptosis.38,57 Yog li ntawd, cov lus ntawm caspase-3, PARP1, thiab cleaved PARP1 hauv MDA-MB-231 hlwb raug tshuaj xyuas los ntawm western blotting. Raws li pom nyob rau hauv daim duab 7, MnPC thiab MnPVA remarkably upregulated qhov kev qhia ntawm caspase-3 thiab cleaved PARP1 raws li piv nrog rau kev tswj thiab lwm yam tebchaw. Qhov kev tshem tawm ntawm PARP1 los ntawm kev qhib lub caspase-3 yuav cais cov DNA-binding domain los ntawm catalytic domain, yog li tiv thaiv PARP ua kom thiab kho cov kab mob DNA. Tsis tas li ntawd, qhov inhibition ntawm PARP1 tuaj yeem nce qib ntawm cov qog-infiltrating T lymphocytes thiab tswj cov lus teb hauv lub cev. DNA-kho tsis ua hauj lwm muaj peev xwm pib lub ntiaj teb DNA aberrations, uas yuav ua rau apoptosis. Proapoptotic Bax thiab antiapoptotic Bcl-xL cov proteins ua lub luag haujlwm tseem ceeb hauv apoptosis.38 Yog li ntawd, peb tau kuaj pom lawv cov lus qhia hauv MDAMB-231 hlwb tom qab kev kho mob nrog cov tshuaj sib txawv rau 48 teev siv western blotting. MnPC thiab MnPVA tau tshaj tawm cov lus qhia ntawm Bax thiab txo qis-tswj cov kev qhia ntawm Bcl-xL. Cov txiaj ntsig tau qhia tias MnPC thiab MnPVA tuaj yeem txhawb nqa apoptosis los ntawm kev tswj hwm apoptotic proteins raws li PARP1- cuam tshuam cov nyhuv cascade. Txawm hais tias VA tuaj yeem ua rau cov qog cell apoptosis los ntawm inhibiting qhov kev qhia ntawm HDAC1/2 ntawm cov koob tshuaj ntau, 58 nws tsis cuam tshuam rau cov proteins apoptotic hauv cov xwm txheej no, qhia txog qhov zoo tshaj ntawm Mn complexes.

Fig. 5 Fluorescence duab ntawm MDA-MB-231 hlwb tom qab incubation nrog MnCl2, MnPC, thiab MnPVA (6 mM) feem rau 36 h thiab staining nrog JC-1 fluorescent sojntsuam.

Ua kom txoj hauv kev cGAS-STING

Nws paub tias PARP1 inhibitor tuaj yeem rub tawm cGAS-STING teeb liab cov lus teb cascade hauv cov qog nqaij hlav cancer.40,59 cGAS yog ib qho kev tiv thaiv kab mob hauv lub cev uas paub txog ntau hom cytosolic dsDNA, suav nrog DNA nrog kab mob, apoptotic, exosomal, mitochondrial, micronuclei. , thiab retroelement origins.60,61 Qhov kev nplua nuj ntawm cytosolic DNA tshwm sim los ntawm MnPC thiab MnPVA tsim ib qho kev pom zoo rau kev ua kom txoj hauv kev cGAS-STING, uas yuav pib pib cov xwm txheej nram qab no los ntawm kev nrhiav neeg ua haujlwm thiab ua kom TBK1 thiab IRF3.12,59 Yog li ntawd, peb tau kuaj pom cov lus qhia ntawm cov proteins no hauv MDA-MB-231 hlwb los ntawm kev tiv thaiv kab mob rau txhua qhov sib xyaw rau 24 teev. Raws li pom nyob rau hauv daim duab 8A, MnPC, thiab MnPVA ho nce qhov kev qhia ntawm cGAS, p-STING, p-TBK1, thiab p-IRF3. Tsis tas li ntawd, peb tau tshawb xyuas qhov cuam tshuam ntawm cov complexes ntawm cGAS-STING txoj hauv kev hauv tib neeg monocytic leukemia THP-1 hlwb los ntawm kev tiv thaiv kab mob. Raws li pom nyob rau hauv daim duab 8B, MnPC thiab MnPVA induced ib tug tseem ceeb upregulation ntawm cGAS, p-STING, p-TBK1, thiab p-IRF3, tshwj xeeb tshaj yog MnPVA, manifests tias lawv qhib lub cGAS-STING txoj kev, uas yuav pib innate kev tiv thaiv los thaiv. qog khiav tawm thiab txhawb kev tiv thaiv kab mob, tsis ua mob rau THP-1 hlwb (Table S3†). Los ntawm qhov sib piv, MnCl2 tsuas yog nce qib ntawm p-STING lossis p-TBK1 thiab tsis tshua muaj kev cuam tshuam rau kev qhia ntawm cGAS thiab p-IRF3 piv nrog rau kev tswj hwm, uas tej zaum yuav yog vim nws tsis muaj peev xwm ua rau muaj kev puas tsuaj DNA thiab PARP1 cleavage ntawm tsawg concentrations. , thiab yog li tsis muaj peev xwm ua rau cov teeb meem qis qis xws li kev ua kom p-IRF3. Raws li cov txiaj ntsig no, peb txiav txim siab tias MnPC thiab MnPVA ua rau muaj kev puas tsuaj rau nuclear thiab mitochondrial DNA hauv cov qog hlwb thiab tso cov DNA tawg mus rau cytoplasm, uas tom qab ntawd qhib txoj hauv kev cGAS-STING. Lub tshuab ua kom cGAS-STING tuaj yeem ncaj qha ua kom tsis muaj zog thiab apoptosis qhia txoj hauv kev hauv cov qog nqaij hlav cancer, 62 ua rau lub cev tiv thaiv kab mob bidirectional. Txawm li cas los xij, qhov kev qhia ntawm STING, TBK1, thiab IRF-3 hauv MDA-MB-231 thiab THP{47}} hlwb tsis tshua muaj kev cuam tshuam (Fig. S8†).

Fig. 6 (A) HDAC kev ua, (B) qhia txog HDAC1/2 cov proteins, thiab (C) cov ntsiab lus ntawm cov protein sib raug rau GAPDH hauv MDA-MB-231 hlwb tom qab incubation nrog txhua qhov sib xyaw (6 mM) rau 48 h txiav txim los ntawm kev siv cov khoom siv ELISA thiab western blotting feem; (D) immunofluorescence dluab ntawm HDAC1 qhia nyob rau hauv 4T1 cov qog nqaij hlav ntawm tus nas tom qab kev kho mob nrog txhua qhov sib xyaw (1.3 mg Mn ib kg) ib zaug txhua 2 hnub rau 16 hnub. *p < 0 05, **p < 0.01, thiab ***p < 0.001.

Interferons thiab proinflammatory cytokines

Kev ua kom TBK1 thiab IRF3 tuaj yeem ua rau muaj kev nthuav tawm thiab tso tawm ntawm IFNs thiab proinflammatory cytokines, xws li IFN-b, TFN-a, thiab IL-6.59,63 Yog li, lub extracellular IFN-I tso tawm los ntawm THP{ {8}} hlwb thiab IFN-b, TNF-a, thiab IL-6 tso tawm los ntawm MDAMB-231 hlwb raug ntsuas los ntawm ELISA kev soj ntsuam ib qho incubation nrog cov tebchaw sib txawv rau 24 teev. Raws li pom nyob rau hauv daim duab 9, MnPC thiab MnPVA ua tau zoo heev txhim kho qhov tso tawm ntawm IFN-I txheeb ze rau kev tswj, MnCl2, thiab VA. Nyob rau tib lub sijhawm, lawv kuj ua rau muaj kev zais ntawm IFN-b thiab TNF-a. Txawm li cas los xij, theem ntawm IL-6 tsuas yog nce siab. IFN-I (IFN-a thiab IFN-b) plays ntau lub luag haujlwm immunostimulatory hauv kev tiv thaiv qog noj ntshav, xws li txhawb kev loj hlob thiab kev nthuav qhia antigen ntawm DCs thiab cov qog nqaij hlav qog nqaij hlav tshwj xeeb los tua cov qog hlwb los ntawm bridging innate thiab adaptive immunity.13 TNF -a yog koom nrog hauv cyclic dinucleotide (CDN)-kev kho mob qog noj ntshav necrosis thiab kho lub cev tiv thaiv kab mob hauv late.59 Cov txiaj ntsig tau qhia tias MnPC thiab MnPVA tuaj yeem txhawb cov tshuaj tiv thaiv kab mob tiv thaiv kab mob thiab qhia tias lawv tuaj yeem kov yeej cov tshuaj tiv thaiv ntawm cov qog los ntawm ib tug chemoimmunotherapeutic mechanism.

Kev loj hlob ntawm pob txha hlwb-derived hlwb (BMDCs)

BMDCs sawv cev rau heterogeneous hlwb ntawm myeloid keeb kwm uas muaj myeloid progenitors thiab tsis paub tab macrophages, granulocytes, thiab dendritic hlwb (DCs); Lawv koom nrog hauv lub cev tiv thaiv kab mob los ntawm suppressing T-cell activation thiab qog-associated macrophage (TAM) activation.64 Raws li lub ntsiab antigen-presenting cells (APCs), DCs ua haujlwm ua tus choj rau kev sib txuas lus ntawm innate thiab adaptive immune systems. 22

Fig. 7 Kev nthuav qhia ntawm DNA kho- thiab apoptosis-hais txog cov proteins hauv MDA-MB-231 cov hlwb tom qab raug rau cov tebchaw sib txawv (6 mM) rau 48 h, thiab cov ntsiab lus ntawm cov protein sib txuas nrog a-tubulin. *p < 0.{{10}}5, **p < 0.01, thiab ***p < 0.001.

Fig. 8 Kev nthuav qhia cov proteins koom nrog hauv cGAS-STING txoj hauv kev txiav txim siab los ntawm sab hnub poob blotting tom qab MDA-MB-231 (A) thiab THP-1 (B) hlwb raug kho nrog 6 thiab 3 mM ntawm ntau cov tebchaw. rau 24 teev, feem, thiab cov ntsiab lus ntawm cov protein sib raug rau GAPDH. *p < 0.{{10}}5, **p < 0.01, thiab ***p < 0.001.

Fig. 9 Kev zais ntawm (A) IFN-I hauv THP-1 hlwb, (B) IFN-b, (C) IL-6, thiab (D) TNF-a hauv MDA-MB{{ 7}} cov hlwb tom qab kev kho mob nrog cov tshuaj sib txawv (6 mM) rau 24 teev. Cov ntaub ntawv qhia tau tias txhais tau tias ± SD (n {{10}}); *p < 0 05, **p < 0.01, thiab ***p < 0.001.

MnPC- thiab MnPVA-activated cGAS-STING txoj hauv kev tuaj yeem ua rau kev loj hlob ntawm BMDCs, peb tau sim cov cim saum npoo ntawm BMDCs los ntawm ow cytometry aer kho nrog cov supernatant ntawm MDA-MB-231 hlwb incubated nrog sib txawv tebchaw rau 24 teev. Raws li pom nyob rau hauv daim duab 10A, piv nrog rau kev tswj, MnPC thiab MnPVA tshwj xeeb tshaj yog ua kom cov co-expression ntawm maturation markers CD86 thiab CD80, ncav cuag 36.62% thiab 43.15%, feem. Txawm li cas los xij, MnCl2 tsuas yog me ntsis dhau ntawm kev tswj hwm. Tsis tas li ntawd, loj histocompatibility complex class II (MHC-II) yog constitutively qhia los ntawm APCs, uas nthuav tawm antigenic peptides rau T hlwb. Raws li qhov tshwm sim, cov lus teb hloov lub cev tiv thaiv kab mob tau pib, tswj hwm, thiab tswj hwm.8 Raws li qhia hauv daim duab 10B thiab C, MnPC, thiab MnPVA tau nce qhov kev qhia ntawm MHC-II txog li 1.5-fold qhov kev tswj hwm . Cov ntaub ntawv qhia tias cov complexes no ua rau muaj kev loj hlob ntawm BMDCs, uas tuaj yeem ua rau lub cev tiv thaiv kab mob ntawm cytotoxic T lymphocytes.

Fig. 10 (A) Co-expression of CD86 and CD80, (B) quantitative analysis of CD11c+CD86+CD80+ in BMDCs, (C) kev qhia ntawm MHC-II ntawm BMDCs 's nto, thiab (D) txhais tau tias kev siv ntawm MHC-II txiav txim siab los ntawm kev ntws cytometry tom qab kev kho mob nrog cov supernatant ntawm MDA-MB-231 hlwb incubated nrog sib txawv tebchaw (6 mM) rau 24 teev.

Kev sib koom ua ke ntawm cov qog nqaij hlav cancer nrog PBMCs

Txhawm rau soj ntsuam ntxiv cov teebmeem immunomodulatory ntawm cov complexes, qhov ua tau zoo ntawm MDA-MB-231 hlwb nyob rau hauv lub xub ntiag ntawm tshuaj-activated tib neeg peripheral ntshav mononuclear hlwb (PBMCs) raug soj ntsuam los ntawm MTT assay raws li ib tug ntaub ntawv txoj kev.66 Cov kab mob qog noj ntshav lossis cov qog nqaij hlav cancer nrog PBMCs tau teeb tsa raws li kev tswj hwm. Raws li qhia hauv daim duab 11, thaum tsis muaj PBMCs, MnPC thiab MnPVA suppressed lub cell viability mus rau 64.98% thiab 62.02%, feem. Kev sib koom ua ke ntawm cov hlwb nrog PBMCs tsis muaj Mn complex tsuas pom basal cytotoxicity nrog lub cev muaj zog ntawm 81.49%. Txawm li cas los xij, nyob rau hauv lub xub ntiag ntawm PBMCs, MnPC thiab MnPVA suppressed cell viability mus rau 38.19% thiab 36.98%, feem. Ntawm qhov concentration (6 mM), feem ntau PBMCs tseem muaj sia nyob (Fig. S9†). Cov txiaj ntsig tau pom tias MnPC thiab MnPVA tuaj yeem qhib lub cev tiv thaiv kab mob ntawm PBMCs kom ua rau muaj kev cuam tshuam cytotoxic ntawm MDA-MB-231 hlwb, yog li nthuav tawm cov txiaj ntsig tseem ceeb ntawm cov qog nqaij hlav cancer.

Mob toxicity thiab nyob rau hauv vivo anticancer kev ua

Qhov mob toxicity ntawm MnII complexes tau soj ntsuam ntawm poj niam Balb/c nas. Qhov koob tshuaj tuag nruab nrab (LD50) thiab lub cev hnyav hloov tom qab txhaj tshuaj ntawm txhua qhov nyuaj tau txiav txim siab. LD50 ntawm MnPC thiab MnPVA yog 16.08 ± 2.16 thiab 25.16 ± 3.19 mg kg−1, raws li (Fig. S10†), uas yog ntau dua qhov ntawd ntawm CDDP (4.{{60}}}6 ± 1.02 mg kg−1 .67 Tsis muaj kev hloov pauv loj hauv lub cev hnyav tau pom hauv MnPC- thiab MnPVA-kho nas. Cov txiaj ntsig tau qhia tias MnPC thiab MnPVA yog cov tshuaj lom neeg tsawg. Interestingly, tethering VA mus rau Mn complex attenuated lub dav toxicity los yog nce lub biocompatibility. Tsis tas li ntawd, cov qauv nas uas muaj 4T1 mob qog noj ntshav tau tsim los ntsuas qhov kev tiv thaiv kab mob ntawm MnPC thiab MnPVA hauv vivo. Raws li pom nyob rau hauv daim duab 12, tom qab kho cov nas nrog PBS, MnCl2, MnPC, thiab MnPVA (1.3 mg Mn ib kg) raws li 16 hnub, MnPC thiab MnPVA pom zoo inhibition ntawm cov qog loj hlob ntau dua MnCl2. Hnub tim 16, qhov nruab nrab ntawm cov qog nqaij hlav (A, B) tau txo qis rau 554.78 ± 43.76 thiab 348.01 ± 39.61 mm3, raws li, rau MnPC- thiab MnPVA-kho nas, thaum ntawd rau PBS- thiab MnCl2- Cov nas kho mob yog 1182.01 ± 95.87 thiab 784 ± 64.93 mm3, feem. Qhov nruab nrab qog hnyav (C) yog 1.41 ± 0.13, 0.95 ± 0.19, 0.82 ± 0.19, thiab 0.59 ± 0.1 g rau PBS-, MnCl2-, MnPC-, thiab MnPVA-kho nas, feem. Obviously, cov qog inhibitory nyhuv ntawm MnPVA yog superior rau ntawm MnPC thiab MnCl2, uas tej zaum yuav yog vim qhov zoo inhibition ntawm HDACs los ntawm MnPVA. Tsis tas li ntawd, tsis pom muaj qhov tshwm sim ntawm qhov txawv txav lossis qhov mob ntawm lub cev qhov hnyav (D), kev ciaj sia, thiab tag nrho lub cev nqaij daim tawv nqaij tau raug soj ntsuam (Fig. S11†). Cov txiaj ntsig no qhia tias MnII complexes tau cog lus rau cov neeg sib tw tshuaj rau kev kho mob qog noj ntshav.

Fig. 11 Qhov nruab nrab muaj peev xwm (%) ntawm MDA-MB-231 hlwb tom qab co-incubation nrog compound-activated PBMCs (6 mM) rau 48 teev. Cov ntaub ntawv qhia tias txhais tau tias ± SD (n=3); **p < 0 01 thiab ***p < 0.001.

Hauv vivo tshuaj tiv thaiv kab mob tiv thaiv kab mob

Lub peev xwm ntawm vivo lub cev tiv thaiv kab mob ntawm MnII complexes tau sim ntawm Balb / c nas bearing 4T1 qog. Cov xwm txheej ntawm lub cev tiv thaiv kab mob hauv cov qog nqaij hlav, spleen, thiab qog-draining lymph nodes (LNs) tau soj ntsuam los ntawm ow cytometry.21 CD8+ T hlwb yog qhov tseem ceeb rau kev txwv kev pib mob qog noj ntshav thiab kev mob qog noj ntshav hauv lub cev, thiab qhov induction ntawm cytotoxic T lymphocytes (CTLs) yuav tsum tau ua kom cov DCs tsis paub qab hau rau cov laus, 8,9 uas tau qhia tias yog CD11c+ CD80+ CD86+ cells.68 Raws li pom hauv daim duab 13A thiab B. , nyob rau hauv MnPVA-kho nas, qhov feem pua ​​​​ntawm cov laus DCs qhia los ntawm kev qhia ntawm co-stimulatory receptors CD80+ thiab CD86+ (ref. 69) yog siab dua (45.59%) hauv LNs tshaj qhov ntawd. kho nrog PBS, MnCl2, thiab MnPC, feem. Macrophages yog ib qho tseem ceeb ntawm lub cev tiv thaiv kab mob, modulating qog microenvironment.70 Raws li cov haujlwm, macrophages tuaj yeem muab faib ua cov qog M1 thiab protumoral M2 phenotypic activation states.71 M1 macrophages raug cim los ntawm TNF-a, IL-12 , CD80, CD86, thiab ncaj qha tua cov qog hlwb, 70 thaum M2-zoo li TAMs yog tus cwj pwm los ntawm kev qhia siab ntawm macrophage mannose receptor 1 (MMR lossis CD206) thiab txhawb kev loj hlob ntawm cov qog hlwb, xws li zoo li nthuav tawm muaj zog immunosuppressive kev ua haujlwm. Nws paub tias inhibitors ntawm HDACs tuaj yeem ua kom muaj zog pro-in-motion shis hauv macrophage polarization thiab txhim kho M1 phenotype ntxiv rau kev tiv thaiv DNA kho.72 Yog li ntawd, peb tau kuaj pom polarization ntawm macrophages hauv tus po thiab qog los ntawm ow cytometry. kev kho mob nrog Mn complexes. Raws li pom nyob rau hauv daim duab 13C–E thiab S12A–C, † lub M2 phenotype cim los ntawm CD206+ tau raug tshem tawm ntau heev thiab M1 phenotype cim los ntawm CD86+ tau nce zoo heev hauv MnPVA-kho nas. Cov txiaj ntsig tau qhia tias MnPVA ua tau zoo induced polarization ntawm macrophages los ntawm M2 mus rau M1 phenotype. MnCl2 thiab MnPC kuj nce qhov kev qhia ntawm CD86, tab sis tsuas yog me ntsis inuenced CD206 piv rau PBS. Lub caij no, IFN-b, IL-6, thiab TNF-ib qho zais cia los ntawm DCs paub tab thiab macrophages raug ntsuas los ntawm ELISA. Raws li pom nyob rau hauv daim duab 13F thiab G, MnPVA txhim kho cov theem ntawm IFN-b thiab TNF-ib qho zoo dua PBS, MnCl2, thiab MnPC, uas tej zaum yuav elicit T hlwb los tua cov qog hlwb. Txawm li cas los xij, tsis pom muaj kev hloov pauv hauv IL-6 (Daim duab S13†).

Daim duab 12 Kev kho cov nyhuv ntawm MnPC, MnPVA, thiab MnCl2 ntawm 4T1 cov qog-cov kabmob nas, siv PBS los tswj. (A) Cov duab ntawm cov qog excised, (B) qog ntim, (C) qog hnyav, thiab (D) lub cev hnyav ntawm cov nas tom qab kho nrog PBS, MnCl2, MnPC, thiab MnPVA, raws li, ntawm 1.3 mg Mn ib kg ib zaug txhua 2 hnub rau 16 hnub. **p < {{10}}.01 thiab ***p < 0.001.

Fig. 13 Mature DCs (CD80+CD86+ gated on CD11c+) thiab quantitative analysis on qog-draining lymph nodes (A thiab B), polarization of macrophages (CD206+CD{ {7}} gated ntawm CD11b+) hauv 4T1 qog nqaij hlav ntawm tus nas (C-E) txiav txim siab los ntawm kev ntws cytometry, thiab ELISA kev soj ntsuam ntawm IFN-b (F) thiab TNF-a (G) hauv cov ntshav los ntawm cov nas tom qab kho nrog txhua qhov sib xyaw (1.3 mg Mn ib kg) ib zaug txhua 2 hnub rau 16 hnub. Cov ntaub ntawv raug nthuav tawm raws li txhais tau tias ± SD (n=3). *P < 0.05; **P < 0.01; ***P <0.001.

Thaum ua kom txoj hauv kev cGAS-STING hauv DCs thiab macrophages los ntawm MnPC thiab MnPVA, cov secreted IFNs-I thiab proinamatory cytokines tuaj yeem ua rau cov cytotoxic lymphocytes.59 Yog li ntawd, peb tau soj ntsuam ntxiv rau kev ua kom cov cytotoxic T hlwb (CD8+ . ) thiab pab T cells (CD4+ ) nyob rau hauv cov qog thiab cov nqaij mos, raws li. Raws li pom hauv daim duab 14A thiab S12D–F†, CD8+ T hlwb (xiav square) hauv cov qog tau ua haujlwm tau zoo (9.18%) hauv MnPVA-kho nas piv rau cov uas tau kho nrog PBS (3.53% ), MnCl2 (4.48%), thiab MnPC (6.05%); CD4+ T hlwb (liab square) kuj tau qhib me ntsis, thiab cov txiaj ntsig tau rov qab rau cov neeg nyob hauv tus po thiab kev kho mob nrog cov complexes. Daim duab 14B qhia tau hais tias cov zaus ntawm cov tshuab txais (CD69+ ) qog-inltrating CD8+ T hlwb tau nce ntau heev. Tag nrho cov txiaj ntsig no qhia tau hais tias MnPVA muaj peev xwm ua kom muaj zog tiv thaiv kab mob hauv vivo vim yog kev ua haujlwm ntawm cGAS-STING txoj hauv kev.

Fig. 14 CD8+ (xiav square) thiab CD4+ T (liab square) cells (A) thiab feem pua ​​ntawm CD69+ subset ntawm CD8+ T cells ( B) hauv 4T1 cov ntaub so ntswg ntawm tus nas tom qab kev kho mob nrog txhua qhov sib xyaw (1.3 mg Mn ib kg) ib zaug txhua 2 hnub rau 16 hnub txiav txim siab los ntawm kev ntws cytometry.

Mechanism ntawm kev ua

kev nqis tes ua rau Mn complexes raws li qhia hauv daim duab 15 tau npaj tseg. Hauv cov qog hlwb, MnPC lossis MnPVA puas nuclear thiab / lossis mitochondrial DNA, ua rau muaj kev tswj hwm ntawm g-H2AX; lub sijhawm no, qhov nyuaj inhibited cov haujlwm ntawm HDAC1/2 thiab PARP1, yog li ua rau lub peev xwm kho DNA thiab ua kom cov DNA puas tsuaj. Cov DNA uas tau sau tseg tau raug tso tawm los ntawm cov nucleus thiab/lossis mitochondria mus rau cytoplasm thiab lees paub los ntawm cGAS los qhib lub STING. Lub STING nyob rau hauv lem stimulated phosphorylation ntawm TBK1 thiab IRF3 thiab translocation rau hauv lub nucleus, inducing zus tau tej cov IFNs, IL-6, thiab TNF-a. Cov IFNs thiab pro-inamatory cytokines tom qab ntawd tau tso tawm los ntawm cov nucleus mus rau cytosol thiab txuas ntxiv mus rau cov qog microenvironment, qhov uas lawv txhawb kev loj hlob thiab kev nthuav qhia antigen ntawm DCs thiab macrophages, thiab txuas ntxiv cytotoxic T hlwb los tua cov qog nqaij hlav cancer. Cov xwm txheej zoo sib xws kuj tau tshwm sim hauv lub cev tiv thaiv kab mob xws li macrophages thiab txoj hauv kev cGAS-STING-TBK1 tau qhib. Kev ua kom txoj hauv kev cGAS-STING tau pib cov lus teb hauv lub cev tiv thaiv kab mob thiab kev sib txuas lus ob txoj hauv kev ntawm cov qog hlwb thiab cov kab mob uas nyob sib ze los txhim kho cov nyhuv tua ntawm cov qog.21 Yog li ntawd, Mn complexes tau pom muaj zog antitumor kev ua ub no vim muaj kev sib koom ua ke ntawm kev kho mob thiab kev tiv thaiv kab mob. . Feem ntau ntawm cov txheej txheem no tau raug pov thawj hauv vitro lossis hauv vivo.

Fig. 15 Cov txheej txheem ntawm kev txiav txim rau manganese complexes.

Xaus

Ntau cov pov thawj qhia tau hais tias ntau cov hlau complexes cuam tshuam nrog cov qog nqaij hlav thiab lub cev tiv thaiv kab mob los kho cov kab mob tiv thaiv kab mob microenvironments ntxiv rau qhov ua rau muaj kev cuam tshuam cytotoxic ncaj qha rau cov qog hlwb.73 Hauv txoj kev tshawb no, peb nthuav tawm ob lub MnII complexes MnPC thiab MnPVA li muaj peev xwm chemo immunotherapeutics los tiv thaiv qog noj ntshav ntawm stimulating. Innate tiv thaiv kab mob thiab ua txhaum DNA ob txoj hlua. Cov multifunctional complexes tua cov qog nqaij hlav cancer tsis yog los ntawm kev puas tsuaj DNA nkaus xwb tab sis kuj los ntawm reactivating lub dormant tiv thaiv teb. Ua ntej, lawv ua raws li DNA breakers, inducing oxidative DNA puas thiab ua DNA tawg; Qhov thib ob, lawv ua raws li inhibitors ntawm HDACs thiab PARP1, tiv thaiv kev kho DNA puas thiab txhawb nqa cov kab mob DNA; thib peb, lawv ua raws li agonists ntawm cGAS-STING txoj kev, inducing lub secretion ntawm IFNs thiab proinflammatory cytokines los txhawb lub maturation ntawm DCs thiab macrophages, thiab ntxiv stimulating qog tshwj xeeb T hlwb los tua cov qog hlwb.

cistanche tubulosa- txhim kho lub cev tiv thaiv kab mob

Nyem qhov no mus saib Cistanche Enhance Immunity khoom

【Nug ntxiv】 Email: cindy.xue@wecistanche.com / Whats App: 0086 18599088692 / Wechat: 18599088692

Txhua yam, MnPC thiab MnPVA ua haujlwm tau zoo rau txoj hauv kev cGAS-STING thiab txwv kev loj hlob ntawm cov qog nqaij hlav hauv vitro thiab hauv vivo. Tethering VA rau MnII complex ua rau nws muaj peev xwm tiv thaiv HDACs thiab ntxiv dag zog rau kev tiv thaiv kev ua haujlwm ntawm lub complex, thiab tsim ntawm Mn complexes muab ib qho tseem ceeb li rau cov kev tiv thaiv proliferative ntawm MnCl2 thiab 1,10-phenanthroline. Txij li thaum Jiang et al.  Ua ntej tshaj tawm txoj haujlwm txhawb nqa ntawm Mn2+ rau txoj hauv kev cGAS-STING, 12 tsis muaj cov cuab yeej zoo li no tau pom hauv Mn complexes. Txoj kev tshawb no qhia tau hais tias lub peev xwm txhawb nqa ntawm MnPC thiab MnPVA rau txoj hauv kev cGAS-STING yog ntau dua li ntawm MnCl2 los yog Mn2+ hauv cov qog thiab lub cev tiv thaiv kab mob vim tias lawv tau ntxias ntau DNA tawg hauv cytoplasm los qhib cGAS. -STING txoj kev. Cov kev tshawb pom qhia tias muaj zog ntau dua cGAS-STING agonists tuaj yeem tau txais los ntawm kev tsim MnII complexes nrog DNA-kev puas tsuaj lossis DNA-kho-blocking ligands.

Cov ntaub ntawv

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