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Chitin plays an important role in the development and molting of insects. The key genes involved in chitin metabolism were considered promising targets for pest control. In this study, two splice variants of chitin deacetylase 2 (CDA2) from Diaphorina citri were identified, including DcCDA2a and DcCDA2b. Bioinformatics analysis revealed that DcCDA2a and DcCDA2b encoded 550 and 544 amino acid residues with a signal peptide, respectively. Spatio-temporal expression patterns analysis showed that DcCDA2a and DcCDA2b were highly expressed in D. citri wing and nymph stages. Moreover, DcCDA2a and DcCDA2b expression levels were induced by 20-hydroxyecdysone (20E). Silencing DcCDA2a by RNA interference (RNAi) significantly disrupted the D. citri molting and increased D. citri mortality and malformation rate, whereas inhibition of DcCDA2b resulted in a semimolting phenotype. Furthermore, silencing DcCDA2a and DcCDA2b significantly suppressed D. citri chitin and fatty acid metabolism. Our results indicated that DcCDA2 might play crucial roles in regulating D. citri chitin and fatty acid metabolism, and it could be used as a potential target for controlling D. citri.
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Citrus , Hemípteros , Animais , Hemípteros/fisiologia , Processamento Alternativo , Quitina , Ácidos GraxosRESUMO
Over the last decade, researchers have found abnormal expression of transient receptor potential (TRP) channels. In particular, members of the thermally sensitive subclass (thermo-TRPs) are involved in many disease processes. Moreover, they have a vital role in the occurrence and development of gastric cancer (GC). Accordingly, thermo-TRPs constitute a major pharmacological target, and the elucidation of the mechanisms underlying their response to physiological stimuli or drugs is key for notable advances in GC treatment. Therefore, this paper summarizes the existing literature about thermo-TRP protein expression changes that are linked to the incidence and progression of GC. The review also discusses the implication of such association to pathology and cell physiology and identifies potential thermo-TRP protein targets for diagnosis and treatment of GC.
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Neoplasias Gástricas , Canais de Potencial de Receptor Transitório , Humanos , Canais de Potencial de Receptor Transitório/metabolismo , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/genéticaRESUMO
Huanglongbing (HLB), also known as citrus greening disease, is caused by Candidatus Liberbacter asiaticus (CLas) and transmitted by Diaphorina citri. Previous studies reported that CLas infection significantly influences the structure of the D. citri cytoskeleton. However, the mechanisms through which CLas manipulates cytoskeleton-related proteins remain unclear. In this study, we performed quantitative ubiquitylome crosstalk with the proteome to reveal the roles of cytoskeleton-related proteins during the infection of D. citri by CLas. Western blotting revealed a significant difference in ubiquitination levels between the CLas-free and CLas-infected groups. According to ubiquitylome and 4D label-free proteome analysis, 343 quantified lysine ubiquitination (Kub) sites and 666 differentially expressed proteins (DEPs) were identified in CLas-infected groups compared with CLas-free groups. A total of 53 sites in 51 DEPs were upregulated, while 290 sites in 192 DEPs were downregulated. Furthermore, functional enrichment analysis indicated that 18 DEPs and 21 lysine ubiquitinated proteins were associated with the cytoskeleton, showing an obvious interaction. Ubiquitination of D. citri tropomyosin was confirmed by immunoprecipitation, Western blotting, and LC-MS/MS. RNAi-mediated knockdown of tropomyosin significantly increased CLas bacterial content in D. citri. In summary, we provided the most comprehensive lysine ubiquitinome analysis of the D. citri response to CLas infection, thus furthering our understanding of the role of the ubiquitination of cytoskeleton proteins in CLas infection.
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Citrus , Hemípteros , Rhizobiaceae , Animais , Proteoma/metabolismo , Proteínas do Citoesqueleto/metabolismo , Tropomiosina/metabolismo , Cromatografia Líquida , Lisina/metabolismo , Espectrometria de Massas em Tandem , Hemípteros/metabolismo , Citoesqueleto/metabolismo , Citrus/metabolismo , Doenças das Plantas/microbiologiaRESUMO
AIM: To investigate whether luteolin, a highly prevalent flavonoid, reverses the effects of epithelial-mesenchymal transition (EMT) in vitro and in vivo and to determine the mechanisms underlying this reversal. METHODS: Murine malignant melanoma B16F10 cells were exposed to 1% O(2) for 24 h. Cellular mobility and adhesion were assessed using Boyden chamber transwell assay and cell adhesion assay, respectively. EMT-related proteins, such as E-cadherin and N-cadherin, were examined using Western blotting. Female C57BL/6 mice (6 to 8 weeks old) were injected with B16F10 cells (1×10(6) cells in 0.2 mL per mouse) via the lateral tail vein. The mice were treated with luteolin (10 or 20 mg/kg, ip) daily for 23 d. On the 23rd day after tumor injection, the mice were sacrificed, and the lungs were collected, and metastatic foci in the lung surfaces were photographed. Tissue sections were analyzed with immunohistochemistry and HE staining. RESULTS: Hypoxia changed the morphology of B16F10 cells in vitro from the cobblestone-like to mesenchymal-like strips, which was accompanied by increased cellular adhesion and invasion. Luteolin (5-50 µmol/L) suppressed the hypoxia-induced changes in the cells in a dose-dependent manner. Hypoxia significantly decreased the expression of E-cadherin while increased the expression of N-cadherin in the cells (indicating the occurrence of EMT-like transformation), which was reversed by luteolin (5 µmol/L). In B16F10 cells, luteolin up-regulated E-cadherin at least partly via inhibiting the ß3 integrin/FAK signal pathway. In experimental metastasis model mice, treatment with luteolin (10 or 20 mg/kg) reduced metastatic colonization in the lungs by 50%. Furthermore, the treatment increased the expression of E-cadherin while reduced the expression of vimentin and ß3 integrin in the tumor tissues. CONCLUSION: Luteolin inhibits the hypoxia-induced EMT in malignant melanoma cells both in vitro and in vivo via the regulation of ß3 integrin, suggesting that luteolin may be applied as a potential anticancer chemopreventative and chemotherapeutic agent.
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Antineoplásicos/uso terapêutico , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Integrina beta3/metabolismo , Neoplasias Pulmonares/prevenção & controle , Luteolina/uso terapêutico , Melanoma Experimental/tratamento farmacológico , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/farmacologia , Western Blotting , Caderinas/biossíntese , Adesão Celular/efeitos dos fármacos , Técnicas de Cultura de Células , Hipóxia Celular , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Imuno-Histoquímica , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/secundário , Luteolina/administração & dosagem , Luteolina/farmacologia , Melanoma Experimental/metabolismo , Melanoma Experimental/patologia , Melanoma Experimental/secundário , Camundongos , Camundongos Endogâmicos C57BL , Invasividade NeoplásicaRESUMO
Genomics is an important subdiscipline of genetics, and it forms a complete research system based on novel theories and techniques. Incorporating genomics in undergraduate curriculum is a response to the need of the development of genetics. The teaching of genomics has significant advantages on developing scientific thinking, enhances bioethics accomplishment, and professional interests in undergraduate students. The integration of genomics into genetics is in accordance with the principles of subject development and education. Related textbooks for undergraduate education are currently available in China, and it is feasible to set up a genetics and genomics integrative course by modifying teaching contents of the genetics course, selecting appropriate teaching approaches, and optimal application of the computer-assisted instruction.
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Genética/educação , Genômica/educação , China , Instrução por Computador , Currículo , HumanosRESUMO
Background: Targeting inflammatory microenvironment is a promising anti-tumor strategy. Paeonol is a phenolic compound with effective anti-inflammatory and anti-tumor properties. However, the effects of paeonol on non-small cell carcinoma (NSCLC) have not been fully investigated. Here, we evaluated the effects of paeonol on proliferation and metastasis of NSCLC and elucidated the underlying mechanisms. Methods: The effects of paeonol on inflammatory cytokines were determined by cell proliferation and ELISA assays. Assays of wound healing, single cell migration and perforation invasion were used to evaluate migration and invasion of NSCLC cells. Expression of marker proteins in epithelial-mesenchymal transition (EMT) and matrix metalloproteinase (MMP) family enzymes were detected by Western blot assays. Nude mouse A549 cells transplantation tumor model was used to study the anti-lung cancer effects of paeonol in vivo. TUNEL stanining were used to detect the apoptosis of tumor cells in A549 lung cancer mice, and Ki67 analysis was used to detect the proliferation of tumor cells in A549 lung cancer mice. Immunohistochemistry was used to detect the effects of paeonol on signaling molecules in tumor tissues. Results: Paeonol inhibited A549 cancer cell migration and invasion in vitro. Paeonol inhibited secreaion of inflammatory cytokines in A549 cells, including tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1ß, and transforming growth factor (TGF)-ß. Paeonol altered the expression of marker proteins involved in EMT and MMP family enzymes. In addition, paeonol inhibited the transcriptional activity of nuclear factor-κB (NF-κB) and phosphorylation of signal transducers and activators of transcription 3 (STAT3). Paeonol inhibited the growth of A549 cells transplanted tumors in nude mice. Conclusion: Paeonol potently inhibited NSCLC cell growth, migration and invasion associated with disruption of STAT3 and NF-κB pathways, suggesting that it could be a promising anti-metastatic candidate for tumor chemotherapy.
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Angiogenesis is crucial for tumor metastasis, with many compounds that inhibit tumor metastasis acting through suppression of angiogenesis. We investigated anti-angiogenic properties of Ligustrazine in a series of in vitro and in vivo models. Ligustrazine inhibited VEGF-induced HUVECs migration and tube formation in a dose-dependent manner in vitro, and had limited cytotoxicity to HUVECs and normal fibroblasts even at a dose up to 100 microg/ml. Ligustrazine also suppressed VEGF-induced rat aortic ring sprouting dose-dependently. Invivo, Ligustrazine reduced the Hb content in a Matrigel plug implanted in mice and inhibited new vessel formation in CAM. In addition, in a B16F10 spontaneous metastasis model, Ligustrazine decreased the expression of CD34 and VEGF in primary tumor tissue and reduced the number of metastasis nodi on the lung surface. Our data suggests that Ligustrazine may inhibit tumor metastasis, at least in part, through its anti-angiogenic activity.
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Inibidores da Angiogênese/uso terapêutico , Neoplasias Pulmonares/prevenção & controle , Melanoma Experimental/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Pirazinas/uso terapêutico , Inibidores da Angiogênese/farmacologia , Animais , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Fibroblastos/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/secundário , Melanoma Experimental/irrigação sanguínea , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos C57BL , Microvasos/efeitos dos fármacos , Neovascularização Patológica/induzido quimicamente , Neovascularização Patológica/patologia , Pirazinas/farmacologia , Ratos , Fator A de Crescimento do Endotélio Vascular/farmacologiaRESUMO
Accumulative evidences have underpinned the nature candidates from Chinese medicine (CM), particularly CM served as blood activating and stasis resolving (BASR, Huoxue Huayu in Chinese) by targeting tumor-associated angiogenesis. However, recent experiment research on the therapeutic angiogenesis by BASR-CM attracts wide attention and discussion. This opinion review focused on the underlying link between two indications and anticipated that (1) BASR-CM might emphasize on a balanced multi-cytokines network interaction; (2) BASR-CM might address on the nature of diseases prior to differently affecting physiological and pathological angiogenesis; (3) BASR-CM might mainly act on perivascular cells, either promotes arteriogenesis by increasing arteriogenic factors in ischemic diseases, or simultaneously keep a quiescent vasculature to impede angiogenesis in tumor context.
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Medicamentos de Ervas Chinesas/uso terapêutico , Neovascularização Patológica/sangue , Neovascularização Patológica/tratamento farmacológico , Inibidores da Angiogênese/química , Inibidores da Angiogênese/uso terapêutico , Animais , Antineoplásicos/uso terapêutico , Medicamentos de Ervas Chinesas/química , Humanos , Modelos BiológicosRESUMO
OBJECTIVE: To study the effect of aqueous extract of several kinds of herbs on human platelet aggregation and expression of P-selectin in vitro. METHODS: Blood was collected from volunteers. Effects of the prepared water extracts of herbs on platelet aggregation were monitored on a Packs-4 aggregometer. The fluorescence intensity of water extracts of Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae on the expression of P-selectin in human platelets of healthy persons was measured with flow cytometry. RESULTS: Out of several herbs investigated, Flos Carthami and Rhizoma Curcumae potently inhibited platelet aggregation after incubation with platelet-rich plasma (PRP) for 15 min. Caulis Spatholobi Flos Carthami and Rhizoma Curcumae inhibited adenosine-5'-diphosphate (ADP) or platelet activating factor (PAF)-induced platelet aggregation in PRP in a dose-dependent manner. In contrast to Flos Carthami and Rhizoma Curcumae, Caulis Spatholobi could not inhibit thrombin-induced platelet aggregation. Despite its inability to inhibit thrombin-induced platelet aggregation in PRP, Caulis Spatholobi had a greater anti-aggregating activity in PRP induced by ADP or PAF. Caulis Spatholobi and Flos Carthami showed significant inhibitory effects on the expression of P-selectin. CONCLUSIONS: Caulis Spatholobi, Flos Carthami and Rhizoma Curcumae have potent anti-platelet properties, and their inhibitory actions are mediated via different mechanisms. Caulis Spatholobi inhibited ADP-induced platelet aggregation but not by thrombin, indicating that its mechanism of action might be independent of the thromboxane pathway. The effect of Caulis Spatholobi and Flos Carthami were associated with suppressing the expression of P-selectin.
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Selectina-P/metabolismo , Extratos Vegetais/farmacologia , Plantas Medicinais/química , Agregação Plaquetária/efeitos dos fármacos , Adulto , Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , Curcuma/química , Fabaceae/química , Humanos , Extratos Vegetais/química , Testes de Função Plaquetária , Água/química , Adulto JovemRESUMO
Pyruvate kinase isozyme type M2(PKM2) was first found in hepatocellular carcinoma(HCC), and its expression has been thought to correlate with prognosis. A large number of studies have demonstrated that epithelial-mesenchymal transition (EMT) is a crucial event in hepatocellular carcinoma (HCC) and associated metastasis, resulting in enhanced malignancy of HCC. However, the roles of PKM2 in HCC EMT and metastasis remain largely unknown. The present study aimed to determine the effects of PKM2 in EGF-induced HCC EMT and elucidate the molecular mechanisms in vitro. Our results showed that EGF promoted EMT in HCC cell lines as evidenced by altered morphology, expression of EMT-associated markers, and enhanced invasion capacity. Furthermore, the present study also revealed that nuclear translocation of PKM2, which is regulated by ERK pathway, regulated ß-catenin-TCF/LEF-1 transcriptional activity and associated EMT in HCC cell lines. These discoveries provide evidence of novel roles of PKM2 in the progression of HCC and potential therapeutic target for advanced cases.
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Carcinoma Hepatocelular/genética , Proteínas de Transporte/genética , Transição Epitelial-Mesenquimal/genética , Receptores ErbB/genética , Neoplasias Hepáticas/genética , Proteínas de Membrana/genética , Hormônios Tireóideos/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Fator de Crescimento Epidérmico/genética , Células Hep G2 , Humanos , Neoplasias Hepáticas/patologia , Fator 1 de Ligação ao Facilitador Linfoide/genética , Prognóstico , Fatores de Transcrição TCF/genética , Transcrição Gênica/genética , beta Catenina/genética , Proteínas de Ligação a Hormônio da TireoideRESUMO
Urea derivatives have been widely used in biology and medicine. The substituted urea derivative URD12 introduced in this study exhibits cytotoxic activity against the K562 human leukemia and KB human mouth epidermal carcinoma cell lines. To further study the bioactivity of URD12 and examine its feasibility as a new antitumor drug, we applied in vivo and in vitro assays to investigate the antitumor activity of URD12. URD12 was prepared and its cytotoxicity was evaluated using the BGC-823 human gastric carcinoma, MGC-803 human gastric carcinoma, SMMC-7721 human hepatoma and HepG2 human hepatocellular carcinoma cell lines using MTT assays. Antitumor activity in vivo was confirmed in mice bearing H22 hepatocellular carcinoma cells. Organ coefficient was used to further elucidate the cytotoxic mechanisms of URD12. URD12 inhibited the growth of tested tumor cell lines in vitro and the growth of H22 mouse hepatocellular carcinoma in vivo with no effects on the weight, spleen and thymus coefficient of tumor-bearing mice. In conclusion, our findings indicate that URD12 is an effective antitumor agent without evident immunosuppression effects.
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OBJECTIVE: To investigate the protective effect of acupuncture intervention on liver in carbon tetrachloride induced hepatic fibrosis rats and to reveal its impact on extracellular matrix production in the liver tissue. METHODS: A total of 46 SD rats were randomly divided into control group (n = 10), model group (n = 12), sham group (n = 12) and acupuncture group (n = 12). Hepatic fibrosis model was established by intraperitoneal injection of 50% olive oil containing CCl (1 mL/kg), 3 times in the 1st week and twice per week from the 2nd to the 6th week. During the fibrosis model establishment, acupuncture of "Taichong" (LR 3), "Qimen" (LR 14), "Ganshu" (BL 18) and "Zusanli" (ST 36) was carried out simultaneously. In the sham group, non-acupuncture points (0.5 cm left to the above-mentioned real points) were punctured. The treatment was conducted 3 times a week in the first three weeks and then twice a week for the last three weeks. Serum levels of hyaluronic acid (HA), liminin (LN) and precollagen (PC III) were determined by enzyme linked immunosorbent assay for assessing the hepatic fibrosis degree. Primary hepatic stellate cells (HSC) were separated. Western blot assay was used to detect the expression of alpha-smooth muscle actin (SMA), alpha 1 (l) collagen, fibronectin, matrix metalloproteinase (MMP)-9 (components of extracellular matrix, ECM) and tissue inhibitor of metalloprotei-nase-1 (TIMP-1, an inhibitor of MMP-9) proteins of HSC. Real-time quantitative polymerase chain reaction was used to detect the expression levels of alpha-SMA, alpha 1 (1) collagen and fibronectin genes of HSC. RESULTS: Compared with the control group, contents of serum HA, LN and PC III, expression levels of alpha-SMA, alpha 1 (I) collagen and fibronectin proteins and genes, and TIMP-1 protein of HSC were significantly increased in the model group (P < 0.01, P < 0.05), while MMP-9 protein (an enzyme for degradating ECM) expression level of HSC in the model group was down-regulated significantly (P < 0.01), suggesting a formation of hepatic fibrosis, impairment of the liver tissue fibrosis and imbalance of degradation of ECM. H.E. staining showed an ameliorated liver injury (disorder of hepatocyte arrangement, hepatocyte necrosis, formation of pseudolobule, etc.) in the acupuncture group in comparison with the model group. In comparison with the model group, serum HA and LN contents, expression levels of alpha-SMA, alpha 1(I) collagen and fibronectin proteins, and alpha-SMA mRNA and fibronectin mRNA of HSC were downregulated considerably in the acupuncture group (P < 0.05, P < 0.01). On the contrary, MMP-9 protein expression level of HSC was up-regulated remarkably in the acupuncture group compared with the model group (P < 0.05). No significant changes were found in all the aforementioned indexes in the sham group, and serum PC III content as well as alpha 1 (I) collagen mRNA and TIMP-1 protein expression levels of HSC in the acupuncture group compared with those in the model group (P > 0.05). CONCLUSION: Acupuncture treatment can significantly relieve CCl4-induced hepatic fibrosis in hepatic fibrosis rats probably by inhibiting the synthesis and deposite of HA and LN, down-regulating expression levels of alpha-SMA, alpha 1(1) collagen and fibronectin proteins, and alpha-SMA mRNA and fibronectin mRNA of HSC as well as up-regulating MMP-9 protein expression of HSC.
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Terapia por Acupuntura , Regulação para Baixo , Matriz Extracelular/metabolismo , Cirrose Hepática/metabolismo , Cirrose Hepática/terapia , Animais , Tetracloreto de Carbono/efeitos adversos , Modelos Animais de Doenças , Humanos , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/enzimologia , Masculino , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Ratos , Ratos Sprague-Dawley , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismoRESUMO
In order to elucidate the toxic mechanisms of Trifolium repens, Festuca arundinacea and Medicago sativa under chromium [Cr (VI)] stress, provide a theoretic foundation for phytoremediation of Cr-contaminated soil, pot experiment was conducted to investigate the effects of Cr(VI) on plant growth, physiological characteristics, Cr accumulation and distribution in three herbaceous plants. Soil sample was treated by adding K2Cr2O7 with the Cr(VI) concentration of 0, 100, 200, 300 and 400 mg x kg(-1), respectively. The results indicated that the average tolerance indices of T. repens, F. arundinacea and M. sativa were 62.5, 48.3 and 36.33, respectively. Compared with control group, contents of chlorophyll, the activity of superoxide dismutase(SOD) and peroxidase (POD) were 57.14%, 51.51%, 35.76% and 63.27%, 52.96%, 41.36% in T. repens, and F. arundinacea, respectively, but M. sativa died in 400 mg x kg(-1) Cr(VI) treatment. The plant height, root length, dry mass of roots and shoots decreased under Cr(VI) stress in three herbaceous plants, and M. sativa > F. arundinacea > T. repens, however, the content of malonyldialdehyde (MDA) increased compared to the control, and the variation range of M. sativa was the highest, while T. repens was the smallest among them. The tolerance of Cr( VI) was T. repens > F. arundinacea > M. sativa. Cr mainly distributed in cell wall and then in the cytoplasm, and less distributed in the mitochondrion and chloroplast in leaves of three herbaceous plants, whereas the content of chlorophyll, MDA, the activity of SOD and POD correlated well with Cr accumulation in the mitochondrion and chloroplast. Cr concentration in the subcellular of leaves increased with the adding Cr(VI) concentration,and M. sativa > F. arundinacea > T. repens. In comparison with T. repens, F. arundinacea, Cr concentration in the leaves of M. sativa was the maximal, i.e. 51.44 mg x kg(-1), and the proportions in the mitochondrion (18.04%) and chloroplast (19.09%) were also higher in 300 mg x kg(-1) Cr(VI). The average accumulation factors of shoots/roots were 1.22/1.54, 1.16/1.44 and 1.26/1.62, while the average translocation factors were 0.78, 0.78 and 0.74 in T. repens, F. arundinacea and M. sativa, respectively. The results suggest that T. repens and F. arundinacea are promising for the phytoremediation of Cr-contaminated soil.
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Cromo/isolamento & purificação , Plantas/metabolismo , Poluentes do Solo/isolamento & purificação , Estresse Fisiológico/fisiologia , Biodegradação Ambiental , China , Cromo/metabolismo , Cromo/toxicidade , Festuca/crescimento & desenvolvimento , Festuca/metabolismo , Medicago sativa/crescimento & desenvolvimento , Medicago sativa/metabolismo , Desenvolvimento Vegetal , Fenômenos Fisiológicos Vegetais , Poluentes do Solo/metabolismo , Poluentes do Solo/toxicidade , Trifolium/crescimento & desenvolvimento , Trifolium/metabolismoRESUMO
The phylogenetic relationships of primates have been extensively investigated, but key issues remain unresolved. Complete mitochondrial genome (mitogenome) data have many advantages in phylogenetic analyses, but such data are available for only 46 primate species. In this work, we determined the complete mitogenome sequence of the black-capped capuchin (Cebus apella). The genome was 16,538 bp in size and consisted of 13 protein-coding genes, 22 tRNAs, two rRNAs and a control region. The genome organization, nucleotide composition and codon usage did not differ significantly from those of other primates. The control region contained several distinct repeat motifs, including a putative termination-associated sequence (TAS) and several conserved sequence blocks (CSB-F, E, D, C, B and 1). Among the protein-coding genes, the COII gene had lower nonsynonymous and synonymous substitutions rates while the ATP8 and ND4 genes had higher rates. A phylogenetic analysis using Maximum likelihood and Bayesian methods and the complete mitogenome data for platyrrhine species confirmed the basal position of the Callicebinae and the sister relationship between Atelinae and Cebidae, as well as the sister relationship between Aotinae (Aotus) and Cebinae (Cebus/Saimiri) in Cebidae. These conclusions agreed with the most recent molecular phylogenetic investigations on primates. This work provides a framework for the use of complete mitogenome information in phylogenetic analyses of the Platyrrhini and primates in general.
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OBJECTIVE: To observe the effect of acupuncture stimulation of "Taichong" (LR 3), "Qimen" (LR 14), etc. on hepatic platelet-derived growth factor (PDGF) signal pathway activity at the protein and mRNA levels in hepatic fibrosis rats. METHODS: Forty-six SD rats were randomly divided into control (10 rats), model (12 rats), acupuncture (12 rats) and non-acupoint (12 rats) groups. Hepatic fibrosis model was established by intraperitoneal injection of mixture solution of 50% CCl4 and olive oil [1:1, 3 times on the 1st week (W), twice/W thereafter for 5 more weeks]. During modeling, acupuncture stimulation of "Taichong" (LR 3), "Qimen" (LR 14), "Ganshu" (BL 18) and "Zusanli" (ST 36) was conducted simultaneously. At the end of the experiments, all the rats were sacrificed for collecting their liver and blood samples, followed by separation of the hepatic stellate cells (HSCs). ELISA, Western blot and Real-time quantitative PCR techniques were used to detect the content of serum PDGF and expression levels of PDGF-beta receptor (PDGF-beta R), extracellular signal-regulated kinase (ERK1/2), c-jun N-terminal kinase (JNK) and P 38 genes and proteins of HSCs, respectively. RESULTS: Compared to the control group, serum PDGF content, and expression levels of PDGF-beta R mRNA and protein, ERK mRNA and protein and P 38 protein of HSCs in the model group were upregulated significantly (P < 0.01, P < 0.05). In comparison with the model group, serum PDGF content, and the expression levels of PDGF-beta R mRNA and protein, ERK mRNA and protein of HSCs in the acupuncture group were down-regulated apparently (P < 0.05, P < 0.01). No significant differences were found between the acupuncture and non-acupoint groups in serum PDGF content and between the model group and non-acupoint group in the expression levels of PDGF-beta R mRNA and protein, ERK mRNA and protein, JNK protein and P 38 protein of HSCs, as well as between the model group and acupuncture group in the expression levels of JNK protein and P 38 protein of HSCs (P > 0.05). CONCLUSION: Acupuncture intervention can effectively down-regulate serum PDGF content, and expression levels of PDGF-beta R mRNA and protein, ERK mRNA and protein of HSCs in liver fibrosis rats, which may contribute to its effect in improving liver fibrosis through down-regulating PDGF signal pathway activity.
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Terapia por Acupuntura , Tetracloreto de Carbono/efeitos adversos , Cirrose Hepática/metabolismo , Cirrose Hepática/terapia , Fator de Crescimento Derivado de Plaquetas/metabolismo , Transdução de Sinais , Animais , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/genética , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/genética , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fator de Crescimento Derivado de Plaquetas/genética , Ratos , Ratos Sprague-Dawley , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/metabolismoRESUMO
Hypoxia-induced epithelial mesenchymal transition (EMT) is an essential step in cancer metastasis. Luteolin, a flavonoid that is widely distributed in plants, is a novel anticancer agent. However, the mechanism underlying its anticancer effects remains undefined. In this study, for the first time, we demonstrate that luteolin inhibits hypoxia-induced EMT in human non-small cell lung cancer cells in culture, which is demonstrated by the fact that hypoxia-induced EMT reduced the expression of E-cadherin and other epithelial markers and increased the expression of N-cadherin, vimentin and other mesenchymal markers; these effects were markedly attenuated by luteolin. In addition, luteolin also inhibited hypoxia-induced proliferation, motility and adhesion in the cells. Furthermore, we reveal that luteolin inhibits the expression of integrin ß1 and focal adhesion kinase (FAK).Since integrin ß1 and FAK signaling are closely related to EMT formation, these results suggest that luteolin inhibits hypoxia-induced EMT, at least in part, by inhibiting the expression of integrin ß1 and FAK.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Neoplasias Pulmonares/patologia , Luteolina/farmacologia , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Adesão Celular/efeitos dos fármacos , Hipóxia Celular , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Transição Epitelial-Mesenquimal/genética , Proteína-Tirosina Quinases de Adesão Focal/genética , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Humanos , Cadeias beta de Integrinas/genética , Cadeias beta de Integrinas/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND: Acupuncture treatment has been increasingly used to treat chronic liver diseases. We previously reported that acupuncture combined with curcumin, a natural antifibrotic compound, could remarkably attenuate liver fibrosis in chemically intoxicated rats, but the underlying molecular mechanisms are poorly understood. The present study was aimed at investigating the effects of acupuncture combined with curcumin on platelet-derived growth factor (PDGF) signalling and extracellular matrix (ECM) regulation in the fibrotic liver. METHODS: A total of 60 Sprague-Dawley male rats were randomly divided into control, model, sham, acupuncture, curcumin and combination treatment groups. During the establishment of fibrosis using carbon tetrachloride (CCl(4)), acupuncture at LR3, LR14, BL18 and ST36 and/or curcumin treatment by mouth were performed simultaneously. After treatment, serum PDGF levels were measured. Protein and mRNA expression of key effectors in PDGF pathway and fibrinolysis in the liver was determined. RESULTS: Acupuncture combined with curcumin potently reduced serum PDGF levels and selectively disrupted the PDGF-ßR/extracellular signal-regulated kinase (ERK) cascade. Combination treatment also significantly repressed expression of connective tissue growth factor and upregulated expression of matrix metalloproteinase-9, promoting fibrinolysis in the fibrotic liver. CONCLUSIONS: The beneficial effects of acupuncture and its combination with curcumin could be attributed to the disruption of PDGF-ßR/ERK pathway and stimulated ECM degradation in the fibrotic liver. Acupuncture treatment significantly enhanced curcumin effects at the molecular level. These findings may provide molecular insights into the potential of acupuncture combined with curcumin for prevention of hepatic fibrosis.
Assuntos
Terapia por Acupuntura , Curcumina/administração & dosagem , Matriz Extracelular/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Cirrose Hepática/terapia , Receptor beta de Fator de Crescimento Derivado de Plaquetas/metabolismo , Animais , Tetracloreto de Carbono/efeitos adversos , Terapia Combinada , MAP Quinases Reguladas por Sinal Extracelular/genética , Humanos , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/genética , Cirrose Hepática/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Transdução de SinaisRESUMO
BACKGROUND: Increasingly, studies demonstrate the effectiveness of acupuncture therapy against liver fibrosis. Curcumin is a natural product with antifibrotic effects, but has poor pharmacokinetic profiles. This study aimed to evaluate whether acupuncture combined with curcumin could more potently attenuate liver fibrosis in chemical intoxicated rats. METHODS: 60 Sprague-Dawley male rats were randomly divided into control, model, sham, acupuncture, curcumin and combination therapy groups. During the establishment of fibrosis using carbon tetrachloride (CCl(4)), acupuncture at LR3, LR14, BL18 and ST36 and/or curcumin treatment by mouth were performed simultaneously. After treatment, pathological indexes and histology for hepatic injury and fibrogenesis were detected. The expression of extracellular matrix (ECM) components was also determined. RESULTS: Acupuncture combined with curcumin potently protected the liver from CCl(4)-induced injury and fibrogenesis, as indicated by reduced levels of serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, hyaluronic acid, laminin and procollagen III. Combined use also led to significant liver histological improvements. Furthermore, combined use effectively inhibited ECM expression such as α-smooth muscle actin, fibronectin and α1(1) collagen. CONCLUSIONS: Acupuncture treatment could significantly enhance the antifibrotic efficacy of curcumin on CCl(4)-induced hepatic fibrosis in rats in vivo, suggesting that a combination of acupuncture with curcumin may be exploited for the prevention of hepatic fibrosis.
Assuntos
Terapia por Acupuntura , Curcumina/uso terapêutico , Cirrose Hepática/terapia , Animais , Tetracloreto de Carbono/efeitos adversos , Terapia Combinada , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Humanos , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/metabolismo , Masculino , Distribuição Aleatória , Ratos , Ratos Sprague-DawleyRESUMO
The present study aimed to investigate the protective effects of injectable caltrop fruit saponin preparation (ICFSP) on ischemia-reperfusion injury in rat brain. Rats were injected with ICFSP and then subjected to cerebral ischemia-reperfusion injury induced by middle cerebral artery occlusion. Then the neurological deficit score was evaluated by Bederson's method. The infarct size was assessed by TTC staining. The content of malondialdehyde (MDA) and nitric oxide (NO), and the activity of superoxide dismutase (SOD) in rat cerebrum were measured with kits, and the content of 6 K prostaglandin F1α (6-K-PGF 1α), thromboxane B2 (TXB2) and endothelin (ET) in blood plasma was measured by radioimmunoassay. The results demonstrated that ICFSP led to a decrease in infarct size (p < 0.01), neurological deficit score (p < 0.05) and plasma content of TXB2 and ET (p < 0.05), and an increase of the plasma level of 6-K-PGF 1α (p < 0.05) and SOD activity in cerebrum, where the MDA and NO content were decreased. The treatment improved forelimb function. ICFSP showed a similar potency compared to that of Ligustrazine hydrochloride parenteral solution (LHPS) and nimodipine (Nim). We concluded that ICFSP protects the brain damage caused by ischemia-reperfusion injury in rats, and this may be closely related to the regulation of reactive oxygen species (MDA and SOD activity) and NO levels in the rat cerebrum, as well as vasoactive factors in the plasma (6-K-PGF 1α, TXB2 and ET).
Assuntos
Infarto Cerebral/prevenção & controle , Cérebro/efeitos dos fármacos , Fármacos Neuroprotetores/uso terapêutico , Fitoterapia , Traumatismo por Reperfusão/prevenção & controle , Saponinas/uso terapêutico , Tromboxano B2 , Tribulus/química , 6-Cetoprostaglandina F1 alfa/sangue , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Bloqueadores dos Canais de Cálcio/farmacologia , Bloqueadores dos Canais de Cálcio/uso terapêutico , Infarto Cerebral/etiologia , Infarto Cerebral/metabolismo , Cérebro/metabolismo , Cérebro/patologia , Endotelinas/sangue , Membro Anterior/efeitos dos fármacos , Membro Anterior/fisiologia , Frutas , Infarto da Artéria Cerebral Média , Injeções , Masculino , Malondialdeído/metabolismo , Fármacos Neuroprotetores/farmacologia , Óxido Nítrico/metabolismo , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Ratos , Ratos Wistar , Traumatismo por Reperfusão/etiologia , Traumatismo por Reperfusão/metabolismo , Saponinas/farmacologia , Superóxido Dismutase/metabolismo , Tromboxano B2/sangueRESUMO
PURPOSE: Cryptotanshinone is a major active component of Salvia miltiorrhiza, which is often used as Chinese herbal medicine in cancer therapy. Here, we systematically assessed the anti-tumor effect of Cryptotanshinone on two melanoma cell lines with low/high-metastatic capacity (B16/B16BL6). METHODS: MTT and LDH assays were used to evaluate cell growth and cytotoxicity. We assessed the effect of Cryptotanshinone on cell apoptosis or proliferation by Annexin V, TUNEL or BrdU assay. Cell cycle distribution was detected by flow cytometry. The integrity of cell cycle checkpoints was determined by mutational analyses of B-RAF and N-RAS, and the expression of cell cycle-associated proteins by western blotting. RESULTS: Treatment with Cryptotanshinone had no obvious effect on cell apoptosis but significantly inhibited cell proliferation. Cryptotanshinone slightly increased the expression of p53, Chk1, and Chk2 in both B16 and B16BL6. Interestingly, Cryptotanshinone induced G1 arrest with a concomitant increase in p21 expression in B16BL6 cells. However, in B16 cells, Cryptotanshinone induced the G2/M arrest through its induction of Cdc25c. Regulation of Cyclin A1, Cyclin B1 and Cdk1/cdc2 expression might contribute to the different cell cycle patterns in B16 and B16BL6 after Cryptotanshinone treatment. CONCLUSIONS: Cryptotanshinone could have diverse effects on cell cycle events in melanoma cell lines with different metastatic capacity. This property might offer an opportunity to study underlying mechanisms for the different antitumor effects of administered Cryptotanshinone in B16 and B16BL6 cells.