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1.
J Ethnopharmacol ; 318(Pt A): 116896, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37437790

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Shrub kava has long been grown and utilized, primarily in the South Pacific region, for ceremonial, religious, and social occasions. It has been used as a pain reliever and muscle relaxant in medicinal practices from the eighteenth century. Interestingly, relatively low incidence of lung cancer may attribute to the high consumption of kava products in this region. AIM OF THE STUDY: Kava extracts were used to produce the kava chalcones Flavokawain A, B and C, which have a variety of bioactivities. In the present study, we show that Flavokawain A has positive effects on endometriosis. MATERIALS AND METHODS: The endometriosis rat model was surgically induced by the autologous transplantation of endometrial tissue. Rats were evaluated for clinical ratings and lesion volume following a 6-week Flavokawain A therapy. Peritoneal fluid and blood samples were taken and ELISA assay was used to measure the cytokines and chemokines levels. Transcriptional and expression levels of Akt, PI3K, NF-kB, iNOS, Bcl-2, Bax and caspase-3 were evaluated by Western blotting and RT-qPCR. Implanted tissue sections of the rats were also analyzed by immunofluorescent and histopathological staining. RESULTS: Lesion volumes and adhesion scores were successfully decreased. Blood and peritoneal fluid levels of associated cytokines and chemokines were markedly down-regulated. Besides, Flavokawain A also mediated cell apoptosis of endometrial implants. Additionally, VEGF expression was reduced, which inhibited the angiogenesis process. As for the expression of Akt, p-Akt, PI3K, p-PI3K, and NF-kB in endometriosis lesions, Flavokawain A significantly reduced them. CONCLUSION: Flavokawain A has beneficial effects on the surgically induced endometriosis rat model, by reducing inflammation, promoting apoptosis, and decreasing angiogenesis. Our findings suggest that these effects may be mediated through the regulation of PI3K/Akt and NF-κB signaling pathways.


Assuntos
Chalcona , Chalconas , Endometriose , Kava , Humanos , Feminino , Ratos , Animais , Chalconas/farmacologia , Chalconas/uso terapêutico , NF-kappa B/metabolismo , Endometriose/tratamento farmacológico , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Chalcona/farmacologia , Citocinas , Apoptose
2.
J Ethnopharmacol ; 318(Pt A): 116901, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37437792

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: The Shenqisherong (SQSR) pill is an empirical prescription of traditional Chinese medicine (TCM), which originated from the National Chinese Medical Science Master, Shi Qi. It has been widely used in the treatment of cervical spondylotic myelopathy (CSM) and promote the recovery of spinal cord function, but underlying molecular mechanism remains unclear. AIM OF THE STUDY: The objective of this study was to confirm the neuroprotective effects of the SQSR pill. MATERIALS AND METHODS: A rat model of chronic compression at double-level cervical cord was used in vivo. The protective role of SQSR pill on CSM rats was measured by Basso, Beattie, and Bresnahan (BBB) locomotor scale, inclined plane test, forelimb grip strength assessment, hindlimb pain threshold assessment, and gait analysis. The levels of reactive oxygen species (ROS) were examined by Dihydroethidium (DHE) staining and 2',7'-Dichlorofluorescein (DCF) assay, and apoptosis was detected by TdT-mediated dUTP nick-end labeling (TUNEL) assay. The expression of apoptosis proteins was evaluated by immunofluorescence staining and Western blot. RESULTS: SQSR pill could facilitate locomotor function recovery in rats with chronic cervical cord compression, reduce local ROS in the spinal cord and downregulate the c-Jun-N-terminal kinase (JNK)/caspase-3 signaling pathway. In addition, the SQSR pill could protect primary rat cortical neurons from glutamate-treated toxicity in vitro by reducing the ROS and downregulating the phosphorylation of JNK and its downstream factors related to neuronal apoptosis meditated by the caspase cascade. Then, the neuroprotective effect was counteracted by a JNK activator. CONCLUSIONS: Together, SQSR pill could ameliorate neuronal apoptosis by restraining ROS accumulation and inhibiting the JNK/caspase-3 signaling pathway, indicating that SQSR pill could be a candidate drug for CSM.


Assuntos
Medula Cervical , Fármacos Neuroprotetores , Traumatismos da Medula Espinal , Ratos , Animais , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio , Medula Cervical/metabolismo , Caspase 3/metabolismo , Transdução de Sinais , Apoptose , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico
3.
J Ethnopharmacol ; 318(Pt A): 116903, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37442494

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: The prescription of Yiqi Jiedu decoction (YQJD) originated from the classic Chinese herbal prescriptions of Danggui Buxue Decoction and Wuzi Yanzong Pill. A previous study has shown that 4 Gy irradiation induced the apoptosis of spermatocytes and revealed autophagosomes in cells exposed to radiation. YQJD decoction has the effect of preventing radiation injury. AIM OF THE STUDY: We used spermatocytes (GC-2spd cell line) to investigate the relationship between autophagy and apoptosis of spermatogenic cells after radiation, and the mechanisms of YQJD decoction. MATERIALS AND METHODS: Establish an in vitro radiation injury model by irradiating GC-2spd cells with 60Co γ-rays (4 Gy or 8 Gy). Autophagy agonists, autophagy inhibitors and YQJD were used to intervene cells. Cell apoptosis and inflammatory factors were measured. NF-κB localization was observed by immunofluorescence. Autophagy and apoptosis-related proteins and IκBα/NF-κB pathway factors were detected. RESULTS: Ionizing radiation promoted the growth of spermatogenic autophagosomes. After radiation, NF-κB was translocated to the nucleus, inflammatory factors were secreted, and IκBα/NF-κB pathway was activated, which promoted autophagy and apoptosis. YQJD decoction can inhibit the phosphorylation of IκBα/NF-κB pathway related factors, regulate the expression of Beclin-1 and Bcl-2 proteins, and inhibit the occurrence of autophagy and apoptosis of irradiated spermatocyte. CONCLUSIONS: The research results indicate that ionizing radiation can activate the IκBα/NF-κB signaling pathway in spermatocytes, promote cell autophagy and apoptosis by regulating the expression of Beclin-1 and Bcl-2 factors. The YQJD decoction inhibits the IκBα/NF-κB signaling pathway so as to regulate Beclin-1 and Bcl-2.


Assuntos
NF-kappa B , Lesões por Radiação , Masculino , Humanos , NF-kappa B/metabolismo , Inibidor de NF-kappaB alfa , Proteína Beclina-1 , Apoptose , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas Reguladoras de Apoptose , Autofagia
4.
J Ethnopharmacol ; 318(Pt A): 116898, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37467820

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Cerebral ischemia-reperfusion injury (CIRI) is a complex pathophysiological process involving multiple factors, and becomes the footstone of rehabilitation after ischemic stroke. Sanpian decoction (SPD) has exhibited protective effects against CIRI, migraine, and other cerebral vascular diseases. However, the underlying mechanisms have not been completely elucidated. AIM OF THE STUDY: This study sought to explore the potential mechanisms underlying the effect of SPD against CIRI. MATERIALS AND METHODS: High-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UPLC) were carried out to determine the chemical constituents of SPD. A network pharmacology approach combined with experimental verification was conducted to elucidate SPD's multi-component, multi-target, and multi-pathway mechanisms in CIRI occurrence. The pharmacodynamics of the decoction was evaluated by establishing the rat model of middle cerebral artery occlusion/reperfusion (MCAO/R). In vivo and in vitro experiments were carried out, and the therapeutic effects of SPD were performed using 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, and Nissl staining. We used terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining and flow cytometry to evaluate cortex apoptosis. The quantification of mRNA and corresponding proteins were performed using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot respectively. RESULTS: Our research showed that pretreatment with SPD improved neurological function and inhibited CIRI. Network pharmacology revealed that the hypoxia-inducible factor-1 (HIF-1) signaling pathway and mitogen-activated protein kinase (MAPK) signaling pathway-mediated apoptosis may be associated with CIRI. In vivo and in vitro experiments, we confirmed that SPD increased cerebral blood flow, improved neural function, and reduced neural apoptosis via up-regulating the expression of sirtuin 1 (SIRT1) and down-regulating phospho-extracellular regulated protein kinases (p-ERK)/ERK and HIF-1α levels in CIRI rats. CONCLUSION: Taken together, the present study systematically revealed the potential targets and signaling pathways of SPD in the treatment of CIRI using in silico prediction and verified the therapeutic effects of SPD against CIRI via ameliorating apoptosis by regulating SIRT1/ERK/HIF-1α.


Assuntos
Isquemia Encefálica , Traumatismo por Reperfusão , Ratos , Animais , Sirtuína 1/metabolismo , Ratos Sprague-Dawley , Isquemia Encefálica/metabolismo , Transdução de Sinais/fisiologia , Apoptose , Infarto da Artéria Cerebral Média/tratamento farmacológico , Infarto da Artéria Cerebral Média/metabolismo , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/metabolismo
5.
J Ethnopharmacol ; 318(Pt A): 116941, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37480970

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Naringenin (NGN) is a widely distributed flavonoid with potent antioxidant and neuroprotective properties. Neuroprotective agents play a crucial role in the treatment of hypoxic-ischemic encephalopathy (HIE). It has shown potential therapeutic effects for neurological disorders. However, its efficacy on HIE is yet to be investigated. AIM OF THE STUDY: This study aims to investigate the potential neuroprotective effect of naringenin and its underlying molecular mechanisms in reducing oxidative stress, apoptosis, and improving brain outcomes following HIE. Additionally, the study aims to identify the potential targets, mechanisms, and functions of naringenin using network pharmacology analysis. MATERIALS AND METHODS: Neonatal mice were exposed to the hypoxic-ischemic brain damage (HIBD) model to determine brain water content, and brain tissue was subjected to hematoxylin and eosin (HE), immunohistochemistry (IHC), terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and Nissl staining to investigate its neuroprotective effects. Furthermore, the neonatal mouse primary neuron oxygen-glucose deprivation (OGD) model to measure reactive oxygen species (ROS) production in vitro. The protein levels were characterized by Western Blot, and mRNA levels were evaluated by a real-time quantitative PCR detecting system (qPCR). Transmission electron microscopy (TEM) and mitochondrial fluorescent staining were used to observe mitochondrial morphology. Neuronal nuclei (NeuN) and microtubule-associated protein 2 (MAP2) were detected by Immunofluorescence (IF). Finally, network pharmacology was employed to determine the common target of naringenin and HIE. The core genes were obtained via protein-protein interaction networks (PPI) analysis and molecular docking was examined, and the mechanism of action was explored through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Additionally, small interfering RNA (siRNA) was constructed for verification. RESULTS: Naringenin has a neuroprotective effect in HIBD by modulating Vegfa expression and activating the PI3K/AKT pathway to inhibit apoptosis. Furthermore, molecular docking results suggest that Vegfa is a potential binding target of naringenin, and silencing Vegfa partially reverses the pharmacological effects of NGN. CONCLUSION: Our findings suggest that naringenin demonstrates potential clinical application for treating HIE as a novel neuroprotective agent.


Assuntos
Hipóxia-Isquemia Encefálica , Fármacos Neuroprotetores , Animais , Camundongos , Hipóxia-Isquemia Encefálica/tratamento farmacológico , Hipóxia-Isquemia Encefálica/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Simulação de Acoplamento Molecular , Apoptose , Encéfalo/metabolismo , Modelos Animais de Doenças , RNA Interferente Pequeno
6.
J Ethnopharmacol ; 318(Pt B): 116976, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37524234

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: The study of male reproductive aging and its associated concerns holds significant importance within the realm of health issues affecting the elderly population. Wubi Shanyao Pills (WSP), a traditional Chinese patent medicine originating from the Tang Dynasty, has been recognized for its ability to enhance male sexual functions while also tonifying the kidney and spleen. Nevertheless, the precise effects and underlying mechanisms through which WSP ameliorates the decline in reproductive function among aging men remain uncertain. AIM OF THE STUDY: This study elucidated the distinctive impacts of WSP on ameliorating the decline in reproductive function caused by natural aging, as well as its underlying mechanisms. MATERIALS AND METHODS: Initially, male mice at the age of 15 months were administered WSP orally at doses of 0.375, 0.75, and 1.50 g/kg per day for a duration of 8 consecutive weeks. The impact of WSP on age-related manifestations in naturally aging mice was assessed based on their behavioral performance. The renal function of the mice was evaluated by measuring serum biochemical indicators, including Creatinine (CR), Uric acid (UA), and Blood urea nitrogen (BUN). Additionally, Superoxide dismutase (SOD) and Malonaldehyde (MDA) levels in renal tissue were determined using applied chemistry methods. Then assessed the levels of Nitric oxide (NO), Total nitric oxide synthase (T-NOS), Guanosine cyclase (GC), and Cyclic guanosine monophosphate (cGMP) in the penile tissue, as well as the expression of Endothelial nitric oxide synthase (eNOS) and Guanylate Cyclase Activator (GUCA) protein, in order to investigate the erectile function of the penis. Additionally, the quality of epididymal sperm was examined using an electron microscope. Furthermore, the serum sex hormone level and related protein expression were determined through the utilization of enzyme-linked immunosorbent assay and immunohistochemistry techniques. Pathological alterations and the ultrastructure of the testis were investigated using hematoxylin-eosin staining and transmission electron microscopy. Subsequently, the apoptosis of spermatogenic cells in the testes was assessed employing TUNEL, immunofluorescence, western blotting, and quantitative real-time polymerase chain reaction. RESULTS: The administration of WSP has been found to enhance the behavioral performance and sexual behavior in aged mice. It's also could increase in serum levels of CR, UA, and BUN, as well as the elevation of SOD activity in kidney tissue, which subsequently leads to a reduction in MDA levels and an improvement in the structural damage caused by aging in the kidney tissue. Consequently, the renal function is enhanced. Additionally, WSP has been observed to elevate the levels of NO, T-NOS, GC, and cGMP in penile tissue, along with an increase in eNOS and GUCA protein expression, indicating an improvement in penile erectile function. The administration of WSP resulted in a decrease in the occurrence of programmed cell death in testicular germ cells, leading to an enhancement in sperm quality and the overall function of testicular spermatogenesis. This improvement can be attributed to the modulation of hormone levels and the regulation of SIRT1/3, p53, FOXO3, Bax, and Caspase-3 expression. CONCLUSION: Collectively, our findings indicate that the administration of WSP has the potential to impede the occurrence of programmed cell death in testicular cells by modulating the expression of SIRT1/3 and subsequent genes associated with apoptosis. Consequently, this regulatory mechanism facilitates the proliferation of testicular cells and sustains the spermatogenic function of the testes. Consequently, by modulating the levels of sexual hormones in naturally aging mice, WSP ultimately enhances the quality of sperm and reproductive function. Concurrently, it also ameliorates age-related behavioral changes, renal function, and erectile function.


Assuntos
Disfunção Erétil , Idoso , Masculino , Camundongos , Humanos , Animais , Disfunção Erétil/tratamento farmacológico , Disfunção Erétil/metabolismo , Medicina Tradicional Chinesa , Sirtuína 1/metabolismo , Sêmen , Testículo , Envelhecimento/fisiologia , Óxido Nítrico Sintase , Superóxido Dismutase/metabolismo , Apoptose
7.
J Ethnopharmacol ; 318(Pt B): 117014, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37557938

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Hemsleya amabilis Diels, belongs to cucurbitaceae, was traditional Chinese medicine (TCM). It is widely used to treat various diseases. However, these diseases may contribute to the development of RCC. AIM OF THE STUDY: investigated the anticancer activities of root extract of Hemsleya amabilis Diels (HRE), and elucidated the underlying molecular mechanism in vivo and in vitro. MATERIALS AND METHODS: Dried Hemsleya amabilis Diels roots were extracted by ethyl acetate and used to treat RCC4, OS-RC-2 and ACHN cells. UHPLC-MS was used to analyze the chemical composition of the extract. CCK-8 and colony formation assay were used to investigate proliferation. PI staining was used to detect cell cycle. Annexin-V-FITC, AO/EB and TEM were used to evaluate apoptosis. Transwell and wound healing assays were used to evaluate migration and invasion. RNA-seq, Network pharmacology, autodocking for virtual screening and molecular dynamics simulation were used to analyze potential molecular mechanisms and active components of HRE inhibiting proliferation of RCC. LY294002 and UC2288 were used to inhibit PI3K and P21 expression, respectively. IGF-1 was used to activate PI3K. Xenograft tumor model was established to evaluate its anti-tumor potential in vivo. Immunohistochemistry and Western blot were used to test protein expression levels. H&E staining was used to explore the side effects of HRE in vivo. Applying bioinformatics to analyze the effect of P21 on RCC. RESULTS: HRE consists of 739 compounds. CCK-8 and colony formation assay showed that HRE significantly inhibited RCC cells proliferation. PI staining indicated that HRE caused G2/M phase arrest. Annexin-V-FITC, AO/EB and TEM experiments revealed that HRE significantly promoted apoptosis of RCC cells. Transwell and wound healing assays showed that HRE can inhibit the migration and invasion of RCC cells. RNA-seq showed that HRE induced 230 gene changes. Network pharmacology analysis found the relationship between HRE-component-target-RCC. Auto-docking found that Epitulipinolide diepoxide in HRE can stably bind to PIK3CA (-7.22 kJ/mol), and molecular dynamics simulation verified the combination between Epitulipinolide diepoxide of PIK3CA. In RCC4 cells, pretreatment with IGF-1, attenuated HRE-induced apoptosis and G2/M arrest. When pretreated with PIK3 inhibitor LY294002, the opposite result appears. Pretreatment with CDKN1A (P21) inhibitor UC2288 attenuated HRE-induced G2/M arrest. Xenograft tumor model showed that HRE inhibited tumor growth. Western blot analysis indicated that HRE can regulating Bax, Bcl-2, PARP, cleared-PARP, Caspase-9, Caspase-8, Caspase-3, Survivin, Cyclin-B1, CDK1, N-cadherin, snail, slug, E-cadherin, MMP-9. Immunohistochemical staining showed that in the treated group, expression of E-cadherin, Bax, P21 was up-regulated, while N-cadherin, PI3K, AKT and Bcl-2 were down-regulated. H&E staining showed that compared to control groups, the main organs in the HRE-treated groups showed no histological abnormalities. The overall survival rate of RCC patients in the high-expression group of P21 was higher than in the low-expression group of P21 on bioinformatics analysis. CONCLUSIONS: HRE inhibited RCC migration and invasion through EMT, and inhibited proliferation in vivo and in vitro. In addition, HRE inhibited proliferation through promoting apoptosis and P21-induced G2/M phase arrest via PI3K/AKT signaling pathway. Overall, these results suggest that HRE may be a promising chemotherapy agent for RCC.


Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Humanos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Carcinoma de Células Renais/tratamento farmacológico , Fosfatidilinositol 3-Quinases/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Fluoresceína-5-Isotiocianato/farmacologia , Inibidores de Poli(ADP-Ribose) Polimerases/farmacologia , Sincalida/metabolismo , Sincalida/farmacologia , Proteína X Associada a bcl-2/metabolismo , Transdução de Sinais , Pontos de Checagem do Ciclo Celular , Apoptose , Proliferação de Células , Neoplasias Renais/tratamento farmacológico , Divisão Celular , Anexinas/metabolismo , Anexinas/farmacologia , Linhagem Celular Tumoral
8.
J Ethnopharmacol ; 318(Pt B): 117025, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37567425

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Polycystic ovary syndrome (PCOS) is a common gynecological endocrine and metabolic disorder. Chinese herbal medicine has some advantages in the treatment of PCOS with its unique theoretical system and rich clinical practice experiences. AIM OF THE STUDY: The present study was to investigate the potential mechanisms of Bu-Shen-Jian-Pi Formula (BSJPF) on the treatment of PCOS. MATERIAL AND METHODS: The combination of ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS/MS) rapid analysis, network pharmacology, molecular docking analysis and bio-experiments were firstly conducted to identify the main effective components of BSJPF, and to predict the potential mechanisms. The ovarian granulosa cell line (KGN) was treated with testosterone to construct the PCOS model in vitro, and the cells were further treated with the lyophilized powder of BSJPF. The levels of proliferation, autophagy and apoptosis were detected to explore the mechanisms of BSJPF on treating PCOS. RESULTS: Firstly, thirty-six active compounds were identified in BSJPF and thirty-one potential targets on PCOS were found. Then, PI3K and PDK1 were verified to have good binding activity with the active compounds through molecular docking analysis. In bio-experiments, BSJPF significantly alleviated the arrested proliferation of KGN cells in G0/G1 phase and reduced the active levels of autophagy and apoptosis of KGN cells induced by testosterone. Additionally, the inhibition of autophagy diminished apoptosis, while the repression apoptosis enhanced autophagy. Finally, BSJPF significantly decreased the FOXO1 expression levels induced by testosterone, especially for nuclear FOXO1, and significantly activated the PI3K/AKT pathway. CONCLUSIONS: BSJPF significantly alleviated the activated autophagy and apoptosis in KGN induced by testosterone through PI3K/AKT1/FOXO1pathway, which is an effective treatment for PCOS.


Assuntos
Medicamentos de Ervas Chinesas , Síndrome do Ovário Policístico , Feminino , Humanos , Testosterona , Medicamentos de Ervas Chinesas/farmacologia , Fosfatidilinositol 3-Quinases , Simulação de Acoplamento Molecular , Síndrome do Ovário Policístico/tratamento farmacológico , Espectrometria de Massas em Tandem , Células da Granulosa , Apoptose , Autofagia , Bussulfano , Proteína Forkhead Box O1 , Proteínas Proto-Oncogênicas c-akt
9.
J Ethnopharmacol ; 318(Pt B): 117054, 2024 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-37595815

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: The Gualou-Xiebai-Banxia decoction (GXBD), a classical traditional Chinese medicine (TCM) formula, has beneficial effects in turbid phlegm obstruction syndrome, a type of coronary heart disease (CHD). However, the underlying mechanism and effective constituents of GXBD remain elusive. Our previous studies have shown that the effective constituents of GXBD may be enriched in the n-butanol fraction (GXB-N) and water fraction (GXB-W), the targets of which remain unknown. AIM OF THE STUDY: To investigate whether GXB-N and GXB-W protect myocardial cells (MCs) via fibroblast growth factor 21 (FGF21) signaling and, if so, to elucidate the underlying mechanisms. Furthermore, to investigate the targets of GXB-N and GXB-W as potential therapeutic targets for cardiovascular disease (CVD). MATERIALS AND METHODS: Cell viability and apoptosis were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays, respectively. The content of FGF21 in the medium was measured using enzyme-linked immunosorbent assay (ELISA). Protein expression was detected using immunofluorescence and western blotting. RESULTS: Apoptosis increased markedly in MCs exposed to oxidized low density lipoprotein (ox-LDL) 100 µg/mL, with increased expression of FGF21, FGFR1 and ßKlotho, phosphorylation of fibroblast receptor substrate 2α (FRS2α) was suppressed. Following incubation with GXB-N and GXB-W 200 µg/mL, the expression of FGF21, FGFR1, and ßKlotho and the phosphorylation of FRS2α were increased. CONCLUSION: Ox-LDL may inhibit the phosphorylation of FRS2α, inducing considerable FGF21 resistance and resulting in MC apoptosis. GXB-N and GXB-W restored and enhanced FGF21 sensitivity in MCs, consequently rescuing cells from ox-LDL-induced apoptosis. The FGF21-FRS2α signal pathway may be part action targets of these two effective fractions of GXBD.


Assuntos
Apoptose , Fatores de Crescimento de Fibroblastos , Transdução de Sinais
10.
Braz. j. biol ; 84: e251336, 2024. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1355879

RESUMO

Abstract Bulbine natalensis and Chorophytum comosum are potential medicinal source for the treatment of cancers. Chronic myeloid leukaemia is a hematopoietic stem cells disorder treated by tyrosine kinase inhibitors but often cause recurrence of the leukaemia after cessation of therapy, hence require alternative treatment. This study determines the anti-cancer effect of leaf, root and bulb methanolic and aqueous extracts of B. natalensis and C. comosum in chronic human myelogenous leukaemia (K562) cell line by MTT, Hoechst bis-benzimide nuclear and annexin V stain assays. The root methanolic extract of B. natalensis and C. comosum showed a high cytotoxicity of 8.6% and 16.7% respectively on the K562 cell line at 1,000 μg/ml concentration. Morphological loss of cell membrane integrity causing degradation of the cell and fragmentation were observed in the root methanolic extract of both plants. A high apoptosis (p < 0.0001) was induced in the K562 cells by both leaf and root extracts of the C. comosum compared to the B. natalensis. This study shows both plants possess apoptotic effect against in vitro myelogenous leukaemia which contributes to the overall anti-cancer properties of B. natalensis and C. comosum to justify future therapeutic applications against chronic myelogenous leukaemia blood cancer.


Resumo Bulbine natalensis Baker e Chorophytum comosum (Thunb.) Jacques são potenciais fontes medicinais para o tratamento de cânceres. A Leucemia Mieloide Crônica (LMC) é um distúrbio das células-tronco hematopoiéticas que é tratado com inibidores da tirosina quinase, mas frequentemente, causa recorrência da leucemia após a interrupção da terapia, portanto, requer um tratamento alternativo. Este estudo determinou o efeito anticancerígeno de extratos metanólicos e aquosos de folha, raiz e bulbo de B. natalensis e C. comosum na linhagem celular de leucemia mieloide humana crônica (K562) por ensaios de MTT, Hoechst bis-benzimida nuclear e anexina V. O extrato metanólico da raiz de B. natalensis e C. comosum apresentou alta citotoxidade de 8,6% e 16,7% respectivamente, na linhagem celular K562 com a concentração de 1,000 μg / ml. Perda morfológica da integridade da membrana celular causando degradação dos núcleos, citoplasma e encolhimento celular foi observada no extrato metanólico da raiz de ambas as plantas. Uma alta apoptose (p <0,0001) foi induzida nas células K562 por extratos de folhas e raízes de C. comosum em comparação com B. natalensis. Este estudo mostrou que ambas as plantas possuem efeito apoptótico contra leucemia mieloide in vitro que contribui para as propriedades anticâncer gerais de B. natalensis e C. comosum para justificar futuras aplicações terapêuticas contra câncer de sangue de LMC.


Assuntos
Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Asphodelaceae , Apoptose , Células K562
11.
Food Chem ; 431: 137154, 2024 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-37595382

RESUMO

The aim of this study was to develop vitamin D3 (VD3) and iron (Fe) blended granules using Neusilin® US2 as an excipient. A central composite design of experiments was used for the continuous manufacturing process, considering VD3 and iron as independent variables and the bulk density, flow index, oil holding capacity, and color difference as response variables. The addition of VD3 had a significant effect on the powder flow properties. The X-ray diffraction and Scanning electron microscopy-energy dispersive X-ray analysis validated the presence of VD3 and Fe in the granules, whereas the variations in porosity and roughness were demonstrated by tomography and atomic force microscopy, respectively. The in vitro cellular uptake profile confirmed the absorption of VD3 in the breast cancer cell line MCF-7 with apparent apoptosis. These results could help in scaling up the process from laboratory to pilot scale in twin-screw granulation and boost the intervention of VitD3/iron deficiencies.


Assuntos
Apoptose , Colecalciferol , Transporte Biológico , Excipientes , Ferro
12.
Clin. transl. oncol. (Print) ; 25(10): 2871-2883, oct. 2023. ilus
Artigo em Inglês | IBECS | ID: ibc-225068

RESUMO

Purpose Doramectin (DRM) is a kind of avermectin drugs, and it has been shown that DRM has anti-cancer effects. However, the molecular mechanism of DRM in programmed cell death (PCD) aspects is still unclear. The objective of this study was to confirm whether DRM induced PCD in glioma cells. Methods In this experiment, the MTT assay and Ki-67 assay were used to detect in vitro cell viability and in vivo tumor proliferation. Then, the effect of DRM on PCD was analyzed by transcriptome comparison. Next, Endogenous apoptosis was detected by transmission electron microscopy (TEM), the DNA gel electrophoresis, JC-1 assay, western blotting and qRT-PCR. Meanwhile, necroptosis was detected by TEM, Hoechst 33342, FITC and PI staining assay, western blotting. Results We found DRM induced apoptosis through Bcl-2/Bax/Caspase-3 pathway. And, DRM induced ROS overproduction, then ROS caused necroptosis through RIPK1/RIPK3/MLKL pathway, Mitochondria acted as a bridge between the two pathways. Conclusion Our research provided new insight with the function of anti-cancer of DRM. These results demonstrated DRM may be used as potential therapeutic agents inducing apoptosis and necroptosis for cancer therapy (AU)


Assuntos
Humanos , Glioma/tratamento farmacológico , Ivermectina/análogos & derivados , Ivermectina/farmacologia , Apoptose/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo
13.
Sci Rep ; 13(1): 15554, 2023 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-37730790

RESUMO

Apoptosis plays a crucial role in cancer pathogenesis and drug resistance. BCL-2 family of enzymes is considered as one of the key enzymes which is involved in apoptosis. When there is disruption in the balance between anti-apoptotic and pro-apoptotic members of the BCL-2 family apoptosis is dysregulated in the affected cells. Herein, 33 novel benzothiazole-based molecules 7a-i, 8a-f, 9a-b, 12a-e, 13a-d, 14a,b, and 17a-j were designed, synthesized and tested for their BCL-2 inhibitory activity. Scaffold hopping strategy was applied in designing of the target compounds. Compounds 13c and 13d showed the highest activity with IC50 values equal to 0.471 and 0.363 µM, respectively. Molecular docking studies of the synthesized compounds showed comparable binding interactions with the lead compound. Structure activity relationship study was performed to show the effects of structural modifications on the inhibitory activities on BCL-2.


Assuntos
Antineoplásicos , Benzotiazóis , Simulação de Acoplamento Molecular , Benzotiazóis/farmacologia , Antineoplásicos/farmacologia , Apoptose , Proteínas Proto-Oncogênicas c-bcl-2
14.
Sci Rep ; 13(1): 15620, 2023 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-37731040

RESUMO

Monoclonal antibodies (mAbs) eliminate cancer cells via various effector mechanisms including antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), which are influenced by the N-glycan structures on the Fc region of mAbs. Manipulating these glycan structures on mAbs allows for optimization of therapeutic benefits associated with effector functions. Traditional approaches such as gene deletion or overexpression often lead to only all-or-nothing changes in gene expression and fail to modulate the expression of multiple genes at defined ratios and levels. In this work, we have developed a CHO cell engineering platform enabling modulation of multiple gene expression to tailor the N-glycan profiles of mAbs for enhanced effector functions. Our platform involves a CHO targeted integration platform with two independent landing pads, allowing expression of multiple genes at two pre-determined genomic sites. By combining with internal ribosome entry site (IRES)-based polycistronic vectors, we simultaneously modulated the expression of α-mannosidase II (MANII) and chimeric ß-1,4-N-acetylglucosaminyl-transferase III (cGNTIII) genes in CHO cells. This strategy enabled the production of mAbs carrying N-glycans with various levels of bisecting and non-fucosylated structures. Importantly, these engineered mAbs exhibited different degrees of effector cell activation and CDC, facilitating the identification of mAbs with optimal effector functions. This platform was demonstrated as a powerful tool for producing antibody therapeutics with tailored effector functions via precise engineering of N-glycan profiles. It holds promise for advancing the field of metabolic engineering in mammalian cells.


Assuntos
Anticorpos Monoclonais , Citotoxicidade Celular Dependente de Anticorpos , Animais , Cricetinae , Anticorpos Monoclonais/genética , Cricetulus , Apoptose , Polissacarídeos/genética
15.
Biol Open ; 12(9)2023 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-37670689

RESUMO

Ubiquitination is a post-translational modification responsible for one of the most complex multilayered communication and regulation systems in the cell. Over the past decades, new ubiquitin variants and ubiquitin-like proteins arose to further enrich this mechanism. Recently discovered ubiquitin variant UbKEKS can specifically target several proteins and yet, functional consequences of this new modification remain unknown. Depletion of UbKEKS induces accumulation of lamin A in the nucleoli, highlighting the need for deeper investigations about protein composition and functions regulation of this highly dynamic and membrane-less compartment. Using data-independent acquisition mass spectrometry and microscopy, we show that despite not impacting protein stability, UbKEKS is required to maintain a normal nucleolar organization. The absence of UbKEKS increases nucleoli's size and accentuate their circularity while disrupting dense fibrillar component and fibrillar centre structures. Moreover, depletion of UbKEKS leads to distinct changes in nucleolar composition. Lack of UbKEKS favours nucleolar sequestration of known apoptotic regulators such as IFI16 or p14ARF, resulting in an increase of apoptosis observed by flow cytometry and real-time monitoring. Overall, these results identify the first cellular functions of the UbKEKS variant and lay the foundation stone to establish UbKEKS as a new universal layer of regulation in the ubiquitination system.


Assuntos
Sistemas CRISPR-Cas , Ubiquitina , Ubiquitina/genética , Ubiquitinas , Ubiquitinação , Apoptose
16.
ACS Appl Mater Interfaces ; 15(37): 43550-43562, 2023 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-37672350

RESUMO

The high temperature induced by surgical electrodes is highly susceptible to severe surface adhesion and thermal damage to adjacent tissues, which is a major challenge in improving the quality of electrosurgery. Herein, we reported a coupled electrode with micro/nano hierarchical structures fabricated by depositing nanoscale hafnium oxide (HfO2) coatings on bionic microstructures (BMs) via laser texturing, acid washing, and atomic layer deposition (ALD) techniques. The synergistic effect of HfO2 coatings and BMs greatly enhanced the hemophobicity of the electrode with a blood contact angle of 162.15 ± 3.16°. Furthermore, the coupled surface was proven to have excellent antiadhesive properties to blood when heated above 100 °C, and the underlying mechanism was discussed. Further experiments showed that the coupled electrode had significant advantages in reducing cutting forces, thermal damage, and tissue adhesion mass. Moreover, the antibacterial rates against Escherichia coli and Staphylococcus aureus were 97.2% and 97.9%, respectively. In addition, the noncytotoxicity levels of HfO2 coatings were verified by cell apoptosis and cycle assays, indirectly endowing the coupled electrode with biocompatibility. Overall, the coupled electrode was shown to have broad potential for application in the field of electrosurgery, and this work could provide new insights into antiadhesion properties under high-temperature conditions.


Assuntos
Biônica , Eletrocirurgia , Antibacterianos/farmacologia , Apoptose , Eletrodos , Escherichia coli , Excipientes
17.
Sci Rep ; 13(1): 14685, 2023 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-37673888

RESUMO

Oncogenic activation of receptor tyrosine kinases (RTKs) such as MET is associated with cancer initiation and progression. We designed and synthesized a new series of quinazoline derivatives bearing 1,2,3-triazole moiety as targeted anticancer agents. The MET inhibitory effect of synthesized compounds was assessed by homogeneous time-resolved fluorescence (HTRF) assay and western blot analysis. Sulforhodamine B assay was conducted to examine the antiproliferative effects of synthetic compounds against 6 cancer cell lines from different origins including MET-dependent AsPC-1, EBC-1 and MKN-45 cells and also Mia-Paca-2, HT-29 and K562 cells. The growth inhibitory effect of compounds in a three-dimensional spheroid culture was examined by acid phosphatase (APH) assay, while apoptosis induction was evaluated by Annexin V/propidium iodide method. Compound 8c bearing p-methyl benzyl moiety on the triazole ring exhibited the highest MET inhibitory capacity among tested agents that was further confirmed by western blot findings. Derivatives 8c and 8h exhibited considerable antiproliferative effects against all tested cell lines, with more inhibitory effects against MET-positive cells with IC50 values as low as 6.1 µM. These two agents also significantly suppressed cell growth in spheroid cultures and induced apoptosis in MET overexpressing AsPC-1 cells. Moreover, among a panel of 24 major oncogenic kinases, the PDGFRA kinase was identified as a target of 8c and 8h compounds. The docking study results of compounds 8c and 8h were in agreement with experimental findings. The results of the present study suggest that quinazoline derivatives bearing 1,2,3-triazole moiety may represent promising targeted anticancer agents.


Assuntos
Apoptose , Receptores Proteína Tirosina Quinases , Anexina A5 , Bioensaio , Western Blotting
18.
Rev. esp. patol ; 56(3): 147-157, Jul-Sep. 2023. tab, ilus, graf
Artigo em Inglês | IBECS | ID: ibc-223319

RESUMO

Introduction: Oral squamous cell carcinoma (OSCC) is the most prevalent head and neck cancer. Few studies have analyzed the expression of proteins related to inflammation (COX-2) and tumor progression according to the histological grade of OSCC. Objective: Analyze the immunohistochemical expression of COX-2, Ki-67 (cell proliferation), Bcl-2/Bax (apoptosis), VEGF, and CD105 (angiogenesis) according to histological grades of OSCC.Material and methodsThe immunohistochemical expression of COX-2, Ki-67, Bcl-2, Bax, VEGF, and CD105 of 58 cases of OSCC was analyzed. 13 cases of oral mucosa (OM) were analyzed as controls. Results: COX-2, VEGF, CD105, and Ki-67 were higher in OSCC than in OM, particularly in poorly differentiated OSCC (p<0.05). Bax expression was lower in poorly differentiated OSCC (p<0.001). The Bcl-2/Bax ratio was higher in OSCC compared to MO (p<0.05). Conclusion: There are immunohistochemical differences according to histological grades of OSCC, which could influence clinical behavior.(AU)


Introducción: El carcinoma oral de células escamosas (COCE) es el cáncer de cabeza y cuello más prevalente. Escasos estudios analizan la expresión de proteínas relacionadas a inflamación (COX-2) y progresión tumoral según el grado histológico de COCE. Objetivo: Analizar la expresión inmunohistoquímica de COX-2, Ki-67 (proliferación celular), Bcl-2/Bax (apoptosis), VEGF y CD105 (angiogénesis) según grados histológicos de COCE.Material y métodos. Se analizó la expresión inmunohistoquímica de COX-2, Ki-67, Bcl-2, Bax, VEGF y CD105 de 58 casos de COCE. Trece casos de mucosa oral (MO) fueron analizados como control. Resultados: Las expresiones de COX-2, VEGF, CD105 y Ki-67 fueron mayores en el COCE comparadas con la MO, particularmente en el COCE pobremente diferenciado (p < 0,05). La expresión de Bax fue menor en el COCE pobremente diferenciado (p < 0,001). La razón Bcl-2/Bax fue mayor en COCE comparado con MO (p < 0,05). Conclusión: Existen diferencias inmunohistoquímicas según grados histológicos de COCE, lo que podría determinar una evolución clínica diferenciada.(AU)


Assuntos
Humanos , Carcinoma , Carcinoma de Células Escamosas , Imuno-Histoquímica , Ciclo-Oxigenase 2 , Proliferação de Células , Apoptose
19.
Dis Model Mech ; 16(9)2023 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-37655466

RESUMO

Epigenetic processes have emerged as important modulators of kidney health and disease. Here, we studied the role of KDM6A (a histone demethylase that escapes X-chromosome inactivation) in kidney tubule epithelial cells. We initially observed an increase in tubule cell Kdm6a mRNA in male mice with unilateral ureteral obstruction (UUO). However, tubule cell knockout of KDM6A had relatively minor consequences, characterized by a small reduction in apoptosis, increase in inflammation and downregulation of the peroxisome proliferator-activated receptor (PPAR) signaling pathway. In proximal tubule lineage HK-2 cells, KDM6A knockdown decreased PPARγ coactivator-1α (PGC-1α) protein levels and mRNA levels of the encoding gene, PPARGC1A. Tubule cell Kdm6a mRNA levels were approximately 2-fold higher in female mice than in male mice, both under sham and UUO conditions. However, kidney fibrosis after UUO was similar in both sexes. The findings demonstrate Kdm6a to be a dynamically regulated gene in the kidney tubule, varying in expression levels by sex and in response to injury. Despite the context-dependent variation in Kdm6a expression, knockout of tubule cell KDM6A has subtle (albeit non-negligible) effects in the adult kidney, at least in males.


Assuntos
Histona Desmetilases , Rim , Feminino , Masculino , Animais , Camundongos , Túbulos Renais , Apoptose , RNA Mensageiro/genética
20.
Artigo em Inglês | MEDLINE | ID: mdl-37659609

RESUMO

Release of agrochemicals from agricultural fields could unintentionally harm organisms that not targeted by pesticides. Flufenacet is one of the oxyacetamide herbicide applied in cultivation fields of crops and this has a possibility of unintentional exposure to diverse ecosystems including streams and surface water. Despite these environmental risks, limited information regarding toxicity of flufenacet on vertebrates is available. This study is aimed to assess environmental hazards and underlying toxic mechanisms of flufenacet by using a zebrafish model. Mortality measurements and morphological observations after the treatment of flufenacet suggested developmental toxicity of flufenacet in zebrafish. In addition, its toxicity on specific organs was evaluated using transgenic fluorescent zebrafish embryo. Adverse effects of flufenacet on vascular and hepatopancreatic development were demonstrated using Tg(flk1:EGFP) and Tg(fabp10a:DsRed; ela3l:EGFP) respectively. To address intracellular actions of flufenacet in zebrafish, cellular responses including apoptosis, cell cycle modulation, and Mapk and Akt signaling pathway were verified in transcriptional and protein levels. These results demonstrated developmental toxicity of flufenacet using the zebrafish model, providing essential information for assessing its potential hazards on vertebrates that are not directly targeted by the pesticide and for elucidating molecular mechanisms.


Assuntos
Fosfatidilinositol 3-Quinases , Peixe-Zebra , Animais , Proteínas Proto-Oncogênicas c-akt , Ecossistema , Fígado , Pâncreas , Apoptose , Transdução de Sinais
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