RESUMO
Neolignans and lignans with diverse new chemical structures, including eleven pairs of separated chiral enantiomers [(+)-/(-)-1-(+)-/(-)-5, (+)-/(-)-8, (+)-/(-)-10, and (+)-/(-)-12-(+)-/(-)-15], two achiral compounds (6 and 9), and an unseparated racemate [(±)-11], together with a new natural product (7) and 21 known derivatives, were isolated from an aqueous extract of the Angelica sinensis root head (guitou). Among the chiral isolates, (+)-/(-)-13 and (+)-/(-)-15 were scalemic pairs with enantiomeric ratios of around 3:1 and 1.5:1, respectively, while others were enantiomeric equivalent pairs. This indicates that the diverse neolignans in A. sinensis are biosynthesized via different pathways with varying degrees of stereo-controlled manners.
Assuntos
Angelica sinensis , Medicamentos de Ervas Chinesas , Lignanas , Lignanas/química , EstereoisomerismoRESUMO
Seven new minor monoterpene derivatives (1-7), together with six known analogues, were isolated from an aqueous decoction of the hook-bearing stems of Uncaria rhynchophylla (Gou-teng). Their structures were determined by spectroscopic data analysis and electronic circular dichroism (ECD) calculations, of which 1 was confirmed by single crystal X-ray diffraction.
Assuntos
Uncaria , Estrutura Molecular , Monoterpenos , Uncaria/química , ÁguaRESUMO
Eleven new sulfonated alkaloids (1 - 11) having diverse structures were isolated from an aqueous extract of the Isatis indigotica root (ban lan gen). Their structures were determined by spectroscopic data analysis, chemical method, and theoretical calculation, of which (-)-4 was proved by single crystal X-ray diffraction.
Assuntos
Alcaloides , Isatis , Alcaloides/química , Isatis/química , Estrutura Molecular , Extratos Vegetais/química , Raízes de Plantas/química , Água/análiseRESUMO
BACKGROUND: Chronic neuropathic pain is a frequent sequel to peripheral nerve injury and maladaptive nervous system function. Divanillyl sulfone (DS), a novel structural derivative of 4,4'-dihydroxydibenzyl sulfoxide from a traditional Chinese medicine Gastrodia elata with anti-nociceptive effects, significantly alleviated neuropathic pain following intrathecal injection. Here, we aimed to investigate the underlying mechanisms of DS against neuropathic pain. METHODS: A chronic constrictive injury (CCI) mouse model of neuropathic pain induced by sciatic nerve ligation was performed to evaluate the effect of DS by measuring the limb withdrawal using Von Frey filament test. Immunofluorescence staining was used to assess the cell localizations and expressions of Iba-1, ASC, NLRP3, and ROS, the formation of autolysosome. The levels of NLRP3-related proteins (caspase-1, NLRP3, and IL-1ß), mitophagy-related proteins (LC3, Beclin-1, and p62), and apoptosis-related proteins (Bcl-XL and Bax) were detected by Western blotting. The apoptosis of BV-2 cell and caspase activity were evaluated by flow cytometry. RESULTS: DS significantly alleviated the neuropathic pain by increasing the mechanical withdrawal threshold and inhibiting the activation of NLRP3 in CCI-induced model mice. Our findings indicated that DS promoted the mitophagy by increasing the LC3II and Beclin 1 and decreasing the levels of p62 protein in BV-2 cell. This is accompanied by the inhibition of NLRP3 activation, which was shown as inhibited the expression of NLRP3 in lysates as well as the secretion of mature caspase-1 p10 and IL-1ß p17 in supernatants in cultured BV-2 microglia. In addition, DS could promote mitophagy-induced improvement of dysfunctional mitochondria by clearing intracellular ROS and restoring mitochondrial membrane potential. CONCLUSION: Together, our findings demonstrated that DS ameliorate chronic neuropathic pain in mice by suppressing NLRP3 inflammasome activation induced by mitophagy in microglia. DS may be a promising therapeutic agent for chronic neuropathic pain.
Assuntos
Inflamassomos/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitofagia/efeitos dos fármacos , Proteína 3 que Contém Domínio de Pirina da Família NLR/efeitos dos fármacos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Neuralgia/tratamento farmacológico , Sulfonas/farmacologia , Sulfonas/uso terapêutico , Animais , Apoptose , Caspase 1/metabolismo , Linhagem Celular , Modelos Animais de Doenças , Inflamassomos/metabolismo , Masculino , Medicina Tradicional Chinesa , Camundongos , Camundongos Endogâmicos C57BL , Microglia/efeitos dos fármacos , Microglia/metabolismo , Microglia/patologia , Mitocôndrias/patologia , Neuralgia/metabolismo , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/metabolismo , Nervo Isquiático/patologiaRESUMO
Seven new monoterpene alkaloids (1-7), along with 18 known analogues, were isolated from an aqueous decoction of the hook-bearing stems of Uncaria rhynchophylla (Gou-teng). Their structures were determined by spectroscopic data analysis and electronic circular dichroism (ECD) calculations. Compound 1 is the first monoterpene 22-norindoloquinolizidine alkaloid with a ketene unit, while 2 and 3 are unusual indoloquinolizidine alkaloids having an oxazinane ring.[Formula: see text].
Assuntos
Alcaloides , Uncaria , Alcaloides Indólicos , Estrutura Molecular , MonoterpenosRESUMO
Five new denudatine-type diterpenoid alkaloids (1-5), along with the known analogue aconicarmine (6), were isolated from an aqueous decoction of the lateral roots of Aconitum carmichaelii (fu-zi). Their structures were determined by spectroscopic data analysis and electronic circular dichroism (ECD) calculations. Compound 5 is the first denudatine-type diterpenoid alcohol iminium alkaloid, which could be partially transformed into the aza acetal form in pyridine-d5. Compound 5 inhibited mice writhing in an acetic acid-induced writhing assay.
Assuntos
Aconitum , Alcaloides , Diterpenos , Animais , Camundongos , Estrutura Molecular , Raízes de PlantasRESUMO
Six new triterpenes, uncarinic acids KP (1-6), along with 24 known analogues, were isolated as minor constituents of an aqueous decoction of the hook-bearing stems of Uncaria rhynchophylla (Gou-teng). By comprehensive spectroscopic data analysis, their structures were elucidated as derivatives of olean-12-en-28-oic acid and urs-12-en-28-oic acid with different oxidized forms at C-3, C-6, and/or C-23, respectively. Cell-based preliminary bioassay showed that the (E)-/(Z)-coumaroyloxy and (E)-/(Z)-feruloyloxy units at C-27 of olean-12-en-28-oic acid and urs-12-en-28-oic acid played roles in their bioactivities.[Formula: see text].
Assuntos
Triterpenos , Uncaria , Estrutura Molecular , Extratos VegetaisRESUMO
Six new indole alkaloid diglycosides named isatigotindolediosides A-F (1-6), along with three known analogs (7-9), were isolated from an aqueous extract of the Isatis indigotica roots (ban lan gen). Their structures including the absolute configurations were determined by comprehensive spectroscopic data analysis, combined with enzyme or acid hydrolysis, and comparison of experimental circular dichroism (CD) and calculated electronic circular dichroism (ECD) spectra. In the preliminary assays, compounds 3, 5, and 8 showed antiviral activity against Coxsackie virus B3.
Assuntos
Glicosídeos/isolamento & purificação , Alcaloides Indólicos/isolamento & purificação , Isatis/química , Raízes de Plantas/química , Antivirais/química , Antivirais/isolamento & purificação , Antivirais/farmacologia , Dicroísmo Circular , Enterovirus Humano B/efeitos dos fármacos , Glicosídeos/química , Glicosídeos/farmacologia , Alcaloides Indólicos/química , Alcaloides Indólicos/farmacologia , Estrutura Molecular , ÁguaRESUMO
Sixteen lignanoids were isolated from an aqueous extract of the commonly used Chinese traditional medicine Dangshen, the dried roots of Codonopsis pilosula, by using a combination of various chromatographic techniques, including silica gel, macroporous adsorbent resin, MCI resin, sephadex LH-20, and reversed phase semi-preparative HPLC. On the basis of spectral data analysis, their structures were elucidated and identified asï¼-ï¼-ï¼7R,7'R,8R,8'Sï¼-4,4'-dihydroxy-3,3',5,5',7-pentamethoxy-2,7'-cyclolignaneï¼1ï¼,ï¼-ï¼-ï¼7R,8Sï¼- dihydrodehydrodiconiferyl alcohol 4-O-ß-D-glucopyranosyl-ï¼1'''â2''ï¼-ß-D-glucopyranosideï¼2ï¼,ï¼-ï¼-ï¼7R,8Sï¼- dihydrodehydrodiconiferyl alcoholï¼3ï¼,ï¼+ï¼-ï¼7S,8Rï¼-dehydrodiconiferyl alcoholï¼4ï¼,ï¼+ï¼-balanophoninï¼5ï¼,ï¼+ï¼- demethoxypinoresinolï¼6ï¼,ï¼+ï¼-pinoresinolï¼7ï¼,ï¼+ï¼-epipinoresinolï¼8ï¼,ï¼-ï¼-syringaresinolï¼9ï¼,ï¼-ï¼-medioresinolï¼10ï¼,ï¼-ï¼-lariciresinolï¼11ï¼,ï¼-ï¼-secoisolariciresinolï¼12ï¼,ï¼-ï¼-ent-isolariciresinolï¼13ï¼,ï¼+ï¼-ï¼7S,8Sï¼-3-methoxy-3',7- expoxy-8,4'-neolignan-4,9,9'-triolï¼14ï¼,ï¼+ï¼-ï¼7S,8Rï¼-3',4-dihydroxy-3-methoxy-8,4'-neolignanï¼15ï¼, andï¼-ï¼-ï¼7R,8Rï¼-3',4-dihydroxy-3-methoxy-8,4'-neolignanï¼16ï¼. All these compounds were isolated from C. pilosula for the first time, while compound 1 is a new natural product of 2,7'-cyclolignan and 2 is a new 4',7-epoxy- 8,3'-neolignan diglucoside. Compound 12 showed activity against Fe(2+)-cysteine induced rat liver microsomal lipid peroxidation with an inhibition ratio ofï¼63.4 ± 8.3ï¼ % at 1×10(-5) mol·L(-1).
Assuntos
Codonopsis/química , Medicamentos de Ervas Chinesas/química , Extratos Vegetais/química , Raízes de Plantas/química , Animais , Butileno Glicóis , Furanos , Lignanas , Microssomos Hepáticos/efeitos dos fármacos , Estrutura Molecular , RatosRESUMO
Seven new C14-polyacetylene glucosides codonopilodiynosides A-G (1-7) were isolated from an aqueous extract of the Codonopsis pilosula roots. Their structures were determined by spectroscopic and chemical methods as (-)-(5S,6E,12E)-tetradeca-6,12-dien-8,10-diyn-1,5,14-triol 5-O-ß-D-glucopyranoside (1), (-)-(5S,6E,12E)-tetradeca-6,12-dien-8,10-diyn-1,5,14-triol 5-O-ß-D-glucopyranosyl-(1â³ â 2')-ß-D-glucopyranoside (2), (-)-(5S,6E,12E)-tetradeca-6,12-dien-8,10-diyn-1,5,14-triol 5,14-di-O-ß-D-glucopyranoside (3), (-)-(5S,6E)-tetradeca-6-en-8,10-diyn-1,5,14-triol 5-O-ß-D-glucopyranoside (4), (-)-(5S,6E,12E)-tetradeca-6,12-dien-8,10-diyn-1,5-diol 5-O-ß-D-glucopyranosyl-(1â³ â 2')-ß-D-glucopyranoside (5), (-)-(6S,4E,12E)-tetradeca-4,12-dien-8,10-diyn-1,6-diol 6-O-ß-D-glucopyranosyl-(1â³ â 2')-ß-D-glucopyranoside (6), and (-)-(5S,6E)-tetradeca-6-en-1,5-epoxy-8,10-diyn-14-ol 14-O-ß-D-glucopyranosyl-(1â³ â 2')-ß-D-glucopyranoside (7), respectively. The absolute configurations of 1-7 were assigned by enzymatic hydrolysis followed by isolation of glucose and aglycones (1a and 4a-7a), and subsequent comparison of specific rotation, TLC, and (1)H NMR data of the glucose with an authentic sugar sample and application of modified Mosher's method based on the MPA determination rule of Δδ(RS) values for 1a and 4a, and Δδ(S) values for 6a. The configuration of 7 was assigned by electronic circular dichroism calculations based on the quantum-mechanical time-dependent density functional theory.
Assuntos
Codonopsis/química , Glucosídeos/isolamento & purificação , Poli-Inos/isolamento & purificação , Glucosídeos/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Raízes de Plantas/química , Poli-Inos/química , EstereoisomerismoRESUMO
Seven new glycosidic bisindole alkaloids, isatindigobisindolosides A-G (1-7), were isolated from an aqueous extract of the Isatis indigotica roots. Their structures including absolute configurations were determined by spectroscopic and chemical methods, together with calculations of electronic circular dichroism (ECD) spectra based on the quantum-mechanical time-dependent density functional theory. In the NMR spectra of 1-3, it is found that integration of H-2 and intensity of C-2 are affected not only by a substitution group at C-2 but also by solvents. Influences of the glucopyranosyloxy on the calculated ECD spectra of the glycosidic bisindole alkaloids are discussed. Compounds 2, 5, and 6 showed antiviral activity against both the influenza virus A/Hanfang/359/95 (H3N2) and Coxsackie virus B3 with IC50 values of 8.4-100.0 µM.
Assuntos
Antivirais/isolamento & purificação , Glicosídeos/isolamento & purificação , Alcaloides Indólicos/isolamento & purificação , Isatis/química , Antivirais/química , Antivirais/farmacologia , Glicosídeos/química , Glicosídeos/farmacologia , Alcaloides Indólicos/química , Alcaloides Indólicos/farmacologia , Vírus da Influenza A Subtipo H3N2/efeitos dos fármacos , Estrutura Molecular , Raízes de Plantas/químicaRESUMO
OBJECTIVE: To analyze the clinical characteristics of the patients with B-cell chronic lymphoproliferative diseaseï¼B-CLPDï¼ in the new drug era and the effect of new drug treatment on efficacy and survival. METHODS: The clinical and laboratory data of 200 cases B-CLPD patients diagnosed between April 2015 and August 2021 were analyzed retrospectively. The clinical efficacy and survival of the patients under different treatments including Bruton tyrosine kinaseï¼BTKï¼ inhibitorsï¼ rituximabï¼ and chemotherapy alone were analyzed. The prognostic factors affecting the survival of patients were analyzed by univarite analysis and multivariate analysis. RESULTS: There were 119 maleï¼59.5ï¼ ï¼ and 81 femaleï¼40.5%ï¼ in 200 cases B-CLPD patientsï¼ the sex ratio(male/female) was 1.5â¶1 with median age of 61ï¼30- 91ï¼ years old. The distribution of subtypes were as fallows: 51 cases (25.5%) of chronic lymphocytic leukemia/small lymphocytic lymphomaï¼CLL/SLLï¼ï¼ 64ï¼32.0%ï¼ cases of follicular lymphomaï¼FLï¼ï¼ 40ï¼20.0%ï¼ cases mantle cell lymphomaï¼MCLï¼ï¼ 30ï¼15.0%ï¼ cases of marginal zone lymphomaï¼MZLï¼ï¼ 10ï¼5%ï¼ cases of lymphoplasmacytic lymphoma/waldenstrom macroglobulinemiaï¼LPL/WMï¼ï¼ 5ï¼2.5%ï¼ cases of B cell chronic lymphoproliferative disorders unclassifiedï¼B-CLPD-Uï¼ . The main clinical manifestation of 102 patients was lymph node enlargement, 32 cases were complicated with B symptoms. Among CLL/SLL patientsï¼ there were 12ï¼23.5%ï¼ cases in Binet A and 39ï¼76.5%ï¼ cases in Binet B/C. There were 29 patientsï¼20.9%ï¼ in Ann Arbor or Lugano stage I-II and 110 casesï¼79.1%ï¼ in stage III-IV of other subtypes. The complete remissionï¼CRï¼ rate was 43.1%ï¼25/58ï¼ï¼ 40.2%ï¼39/97ï¼ï¼ 7.1%ï¼1/14ï¼ï¼ and overaIl response rateï¼ORRï¼ was 87.9%ï¼51/58ï¼ï¼ 62.9%ï¼61/97ï¼ï¼ 28.6%ï¼4/14ï¼ in the groups of BTK inhibitorsï¼ rituximab-based therapyï¼ and chemotherapy alone. The 3-year OS rate and PFS rate in all patients was 79.2% and 72.4% respectively. The 3-year OS rate of patient with MZL, CLL/SLL, FL,WM was 94.7%, 87.7%, 86.8% and 83.3% respectively, while the 3-year OS rate of MCL was only 40.6%, which was significantly lower than other subtypes. The median OS of patients treated with BTK inhibitors and rituximab-based therapy was 20.5 and 18.5 months respectivelyï¼ and the 3-year OS rate was 97.4% and 90.7%. Howeverï¼ the median PFS of patients receiving chemotherapy alone was 4 monthsï¼ and the 1-year OS rate was 52.7%ï¼ which was statistically significant compared with the other two groupsï¼P<0.05ï¼. Univarite analysis showed that anemiaï¼ elevated lactate dehydrogenaseï¼ elevated ß2-microglobulinï¼ and splenomegaly were the poor prognostic factors for OSï¼P<0.05ï¼ï¼ elevated lactate dehydrogenase was also poor prognostic factors for PFSï¼P<0.05ï¼. Multifactor analysis showed that anemia and elevated lactate dehydrogenase were the independent poor prognostic factors for survivalï¼P<0.05ï¼. CONCLUSION: The clinical features of B-CLPD was variousï¼ anemia and elevated lactate dehydrogenase are the prognostic factors for poor survival. BTK inhibitors and new immunotherapy can improve the survival and prognosis of patients in the new drug era.
Assuntos
Leucemia Linfocítica Crônica de Células B , Linfoma de Zona Marginal Tipo Células B , Linfoma de Célula do Manto , Humanos , Adulto , Feminino , Masculino , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Rituximab/uso terapêutico , Leucemia Linfocítica Crônica de Células B/tratamento farmacológico , Estudos Retrospectivos , Prognóstico , Lactato DesidrogenasesRESUMO
OBJECTIVE: To investigate the effect of miR-203/CREB1 signaling regulation mediated by DNA methylation on the proliferation, invasion and apoptosis of multiple myeloma (MM) cells. METHODS: The methylation level of miR-203 in the RPMI 8226 cells was detected by bisulfite sequcucing polymerase chain reaction (BSP). The mRNA expression of miR-203 was measured by quantitative real-time polymerase chain reaction. RPMI 8226 cells were treated with DNA methyltransferase inhibitor 5-Aza-2'-deoxycytidine (5-Aza-CdR). The miR-203 mimic in MM cell line RPMI 8226 was transfected to establish overexpressed miR-203 cell. The proliferation, invasion ability and apoptosis of RPMI 8226 cell was detected by CCK-8 assay, Transwell, and flow cytometry, respectively. The targeting relationship between miR-203 and CREB1 was verified by double luciferase report assay. Western blot was used to detect the expression of CREB1 protein. RESULTS: Hypermethylation of miR-203 promoter region and low expression level of miR-203 mRNA were detected in the RPMI 8226 cells, which showed that demethylation could induce the expression of miR-203. The proliferation and invasion ability of RPMI 8226 cells after treated by 5-Aza-CdR were inhibited, and showed statistical significance as compared with blank control group (both P<0.05)ï¼while the apoptosis rate was promoted (P<0.05). The proliferation, invasion ability and apoptosis of overexpressed miR-203 were the same as the demethylation group. Double luciferase report assay confirmed that CREB1 was the direct target of miR-203. The protein level of CREB1 was inhibited by demethylation and showed statistical significance as compared with control group (P<0.05). CONCLUSION: MiR-203 targeting CREB1 mediated by DNA methylation leads to maintain the malignant biological behaviors of MM cells.
Assuntos
MicroRNAs , Mieloma Múltiplo , Apoptose , Azacitidina/farmacologia , Linhagem Celular Tumoral , Proliferação de Células , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/farmacologia , Metilação de DNA , Regulação Neoplásica da Expressão Gênica , Humanos , MicroRNAs/metabolismo , Mieloma Múltiplo/genética , RNA Mensageiro/metabolismoRESUMO
OBJECTIVE: To investigate the relationship between promoter hypermethylation of secreted frizzled-related protein (SFRP) gene and acute leukemia (AL). METHODS: We examined the promoter methylation status of SFRP1, 2, 4 and 5 in primary or relapsed AL patients, cell lines (HL60, NB4, Molt-4 and Jurkat) and peripheral blood mononuclear cells from healthy people with methylation-specific PCR (MSP). RESULTS: None of the normal mononuclear cells showed methylation of any SFRP genes. The frequencies of aberrant methylation among the samples were 33.9% (20/59) for SFRP1, 23.7% (14/59) for SFRP2, 6.8% (4/59) for SFRP4 and 10.2% (6/59) for SFRP5 in acute myelocytic leukemia (AML), and 39.3% (11/28) for SFRP1, 28.6% (8/28) for SFRP2, 25.0% (7/28) for SFRP4 and 32.1% (9/28) for SFRP5 in acute lymphoblastic leukemia (ALL). Hypermethylation of SFRP1, 2, 5 genes were present in the 4 AL cell lines. SFRP4 was methylated in NB4, Molt-4 and Jurkat cell lines. However, methylation and unmethylation of SFRP4 were both detected in HL60. CONCLUSIONS: Hypermethylation of SFRP genes is a common event in the evolution of AL. Methylation of SFRP genes might serve as potential independent biomarkers for early detection of AL.
Assuntos
Metilação de DNA , Receptores Frizzled/metabolismo , Leucemia/metabolismo , Regiões Promotoras Genéticas , Doença Aguda , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Linhagem Celular Tumoral , Feminino , Receptores Frizzled/genética , Inativação Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Leucemia/genética , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Transdução de Sinais , Adulto JovemRESUMO
Two new folate-derived analogues, named uncarophyllofolic acids A (1) and B (2), respectively, were isolated from the Uncaria rhynchophylla hook bearing stem (Gouteng in Chinese). The distinct stereochemical structures of 1 and 2 were determined by spectroscopic data analysis in combination with acidic hydrolysis and Marfey's derivatization, along with comparison of their specific rotation and Cotton effect (CE) data with those of the biogenetically related known derivatives as well as theoretical calculations of electronic circular dichroism (ECD) spectra. A plausible biosynthetic pathway of 1 and 2, associating to folate metabolism and the previously reported orychophragines A-C from Orychophragmus violaceus, is discussed.
Assuntos
Ácido Fólico/química , Extratos Vegetais/química , Uncaria/química , China , Cromatografia Líquida de Alta Pressão , Ácido Fólico/análogos & derivados , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Caules de Planta/químicaRESUMO
Nine new gastrodin derivatives, including seven p-hydroxybenzyl-modified gastrodin ethers (1-7), 6'-O-acetylgastrodin (8), and 4-[α-D-glucopyranosyl-(1 â6)-ß-D-glucopyranosyloxy]benzyl alcohol (9), together with seven known derivatives, were isolated from an aqueous extract of Gastrodia elata ("tian ma") rhizomes. Their structures were determined by spectroscopic and chemical methods as well as single crystal X-ray diffraction. Compounds 1-4, 7, 10, and 11 were also isolated from a reaction mixture by refluxing gastrodin and p-hydroxybenzyl alcohol in H2O. As both gastrodin and p-hydroxybenzyl alcohol exist in the plant, the reaction results provide evidence for the production and increase/decrease of potential effective/toxic components when "tian ma" is decocted solely or together with ingredients in Chinese traditional medicine formulations, though the isolates were inactive in the preliminarily cell-based assays at concentrations of 10 µM. Moreover, using ultra-performance liquid chromatography high-resolution electrospray ionization mass spectrometry (UPLC-HRESIMS), 4, 7, 10, and 11, as well as component variations, were detectable in the freshly prepared extracts of different types of samples, including the freeze-dried fresh G. elata rhizomes.
RESUMO
OBJECTIVE: To investigate the effect of triptolide(TPL) on proliferation and apoptosis of RPMI8226 cells and its mechanism. METHODS: MTT assay was used to measure the proliferation of RPMI8226 cells after treatment with different concentration (10, 20, 40, 80 and 160 nmol/L) of TPL for different incubation time (24 h, 48 h and 72 h). The cell apoptosis was detected by flow cytometry, the mRNA expressions of SMYD3 and MMP-9 were measured by quantitative real-time PCR, the protein level of H3K4me2 and H3K4me3 in RPMI8226 cells was assayed by Western blot. RESULTS: TPL inhibited RPMI8226 cell proliferation, and the inhibitory rate of cell proliferation increased significantly in a dose- and time-dependent manner(P<0.05), the RPMI8226 cell apoptosis was induced by treatment with 40, 80 and 160 nmol/L TPL (P<0.05), the qRT-PCR showed that treatment of RPMI8226 cells with TPL down-regulated the mRNA expression of SMYD3 in a dose-dependent manner(P<0.05). Compared with the blank group, the mRNA expression level of MMP-9 in RPMI8226 cells transfected by siRNA-SMYD3 was significantly depressed. Western blot showed that the protein levels of H3K4me2 and H3K4me3 were decreased in a dose-dependent manner after TPL treatment(P<0.05). Compared with the blank group and siRNA negative group, the protein level of H3K4me2 and H3K4me3 in RPMI8226 cells transfected by siRNA-SMYD3 also were significantly depressed(P<0.05). CONCLUSION: TPL can significantly inhibit the proliferation of RPMI8226 cells and induce their apoptosis, which may be related to the inhibition of SMYD3 expression by TPL- down-regulating the H3K4 methylation and the activating the MMP-9 transcription.
Assuntos
Mieloma Múltiplo , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Diterpenos , Compostos de Epóxi , Histona-Lisina N-Metiltransferase , Humanos , Metaloproteinase 9 da Matriz , FenantrenosRESUMO
OBJECTIVE: To investigate the effects of triptolide (Chinese Traditional Medicine)on the apoptosis and H3K4 protein methylation in multiple myeloma cells. METHODS: The RPMI8226 cells were cultured with different concentrations(10,20,40,80 and 160 nmol/L)of triptolide for different incubation time (24 h,48 h and 72 h). The inhibition of triptolide on RPMI8226 cell proliferation was detected by MTT assay. Apoptosis and cell cycle distribution were evaluated by flow cytometry.The expressions of H3K4me2 and trimethylation of histone H3 lysine 4(H3K4me3) in RPMI8226 cells were assayed by Western blot. The changes of expressions of histone methylase SET and MYND domain containing 3(SMYD3) and histone demethylase lysine specific demethylase 1(LSD1) in RPMI8226 cells were verified by qRT-PCR. RESULTS: Triptolide had obvious inhibitive effects on proliferation of RPMI8226 cells and showed a dose-and time-dependent manner(P<0.05). Triptolide induced apoptosis and G2/M cell cycle arrest in a dose-dependent manner(P<0.05). Triptolide decreased histone H3K4me2 and H3K4me3 expression in a dose-dependent manner(P<0.05, P<0.01). SMYD3 was significantly depressed at protein expression in a dose-dependent manner(P<0.05), but LSD1 was up-regulated (P<0.05). CONCLUSIONS: Triptolide could inhibit RPMI8226 cell proliferation,induce the apoptosis and cause G2/M arrest,meanwhile,significantly inhibit the protein expressions of H3K4me2 and H3K4me2 with alter the expression of SMYD3 and LSD1.The effects is probably related to the antitumor mechanism of MM cells induced by triptolide.
Assuntos
Apoptose/efeitos dos fármacos , Diterpenos/farmacologia , Histona-Lisina N-Metiltransferase/metabolismo , Histonas/metabolismo , Mieloma Múltiplo/metabolismo , Fenantrenos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células , Compostos de Epóxi/farmacologia , Histona Desmetilases/metabolismo , Humanos , MetilaçãoRESUMO
OBJECTIVE: To investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs). METHODS: HRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA. RESULTS: The protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG. CONCLUSION: The expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.
Assuntos
Quimiocina CCL2/metabolismo , Produtos Finais de Glicação Avançada/farmacologia , Células Mesangiais/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Soroalbumina Bovina/farmacologia , Células Cultivadas , Humanos , Células Mesangiais/efeitos dos fármacos , Receptor para Produtos Finais de Glicação Avançada , Receptores Imunológicos/metabolismoRESUMO
BACKGROUND: DNA methylation of CpG islands within the promoters of specific genes may play roles in tumor initiation and progression. It has been suggested such events may serve as critical check points. METHODS: The present study analyzed the methylation status of CpG islands within the promoters of secreted frizzled-related proteins (SFRPs) in 87 acute leukemia (AL) patients, 20 normal controls, and four AL cell lines. 5-aza-2'- deoxycytidine (5-Aza-CdR), an inhibitor of DNA methylation, was employed to determine its effect on SFRP expression. RESULT: Methylation of at least one SFRP promoter was observed in 69% of the AL patients analyzed. In addition, methylation of all four SFRP promoters was observed in Molt-4, Jurkat, HL60 and NB4 cells. In Jurkat cells, methylation levels of four SFRP promoters decreased in a dose-dependent manner upon treatment with 5-Aza-CdR, which coincided with increased mRNA expression. With increasing 5-Aza-CdR concentrations, the expression of DNA methyltransferases, DNMT3A and DNMT3B, significantly decreased in a dose-dependent manner. CONCLUSION: The present study demonstrated that SFRP gene methylation may be involved in AL progression, with a possible epigenetic mechanism influencing Wnt signaling.