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1.
São Paulo; s.n; s.n; 2022. 172 p. tab, graf.
Thesis in English | LILACS | ID: biblio-1378625

ABSTRACT

The solar ultraviolet (UV) radiation that reaches the Earth is composed of 95% of UVA (320 to 400 nm) and 5% of UVB (280 to 320 nm) radiation. UVB is carcinogenic, generating potentially mutagenic DNA lesions. The solar UVA radiation also causes DNA damage, but this fact does not fully account for its biological impact. UVA is absorbed by non-DNA cellular chromophores, generating reactive oxygen species such as singlet oxygen. Knowing the proteome mediates stress responses in cells, here we investigated the cellular effects of a non-cytotoxic dose of UVA radiation, equivalent to about 20 minutes of midday sun exposure, on the proteome of human keratinocytes. Using a combination of mass spectrometry-based proteomics, bioinformatics, and conventional biochemical assays, we analyzed two aspects of UVA-induced stress: spatial remodeling of the proteome in subcellular compartments 30 minutes after stress and long-term changes in protein levels and secretion (24 hours and 7 days postirradiation). In the first part of this thesis, we quantified and assigned subcellular localization for over 3000 proteins, of which about 600 potentially redistribute upon UVA exposure. Protein redistributions were accompanied by redox modulations, mitochondrial fragmentation and DNA damage. In the second part of the work, our results showed that primary human keratinocytes enter senescence upon exposure to a single dose of UVA, mounting antioxidant and inflammatory responses. Cells under UVA-induced senescence further elicit paracrine responses in neighboring premalignant HaCaT epithelial cells via inflammatory mediators. Altogether, these results reiterate the role of UVA radiation as a potent metabolic stressor in the skin


A radiação ultravioleta (UV) solar que atinge a superfície terrestre é composta por 95% de radiação UVA (320 a 400 nm) e 5% de radiação UVB (280 a 320 nm). A radiação UVB é carcinogênica e gera lesões potencialmente mutagênicas no DNA. A radiação UVA solar também gera danos no DNA, mas a genotoxicidade dessa radiação não explica inteiramente o seu impacto biológico. Atualmente, sabe-se que a radiação UVA é absorvida por cromóforos celulares, gerando espécies reativas de oxigênio, como o oxigênio singlete. Sabendo que o proteoma é um mediador de respostas ao estresse celular, nós investigamos os efeitos celulares de uma dose não-citotóxica de radiação UVA, equivalente a cerca de 20 minutos de exposição ao sol, no proteoma de queratinócitos humanos. Utilizando espectrometria de massas, bioinformática e ensaios bioquímicos convencionais, nós analisamos dois aspectos do estresse induzido por radiação UVA: o remodelamento espacial do proteoma 30 minutos depois do estresse e alterações nos níveis e na secreção de proteínas no longo prazo (24 horas e 7 dias depois da irradiação). Na primeira parte desta tese, nós quantificamos e atribuímos classificações de localização subcelular a mais de 3000 proteínas. Dentre essas proteínas, 600 tem potencialmente a sua distribuição subcelular alterada em resposta à radiação. As redistribuições subcelulares são acompanhadas de modulações redox, fragmentação mitocondrial e danos no DNA. Na segunda parte da tese, os nossos resultados mostraram que queratinócitos humanos primários entram em senescência sob exposição a uma única dose de radiação UVA, montando respostas antioxidantes e pró-inflamatórias. Células sob senescência induzida por UVA, por sua vez, desencadeiam respostas parácrinas em queratinócitos pré-tumorais (células HaCaT) por meio de mediadores inflamatórios. Em conjunto, esses resultados reiteram o papel da radiação UVA como um potente estressor metabólico em células da pele


Subject(s)
Skin , Ultraviolet Rays/adverse effects , Keratinocytes/chemistry , Proteomics/classification , Radiation Dosage , Mass Spectrometry/methods , DNA , Epithelial Cells/classification , Genotoxicity/adverse effects , HaCaT Cells/classification , Antioxidants/adverse effects
2.
Journal of Medical Biomechanics ; (6): E004-E017, 2022.
Article in Chinese | WPRIM | ID: wpr-920662

ABSTRACT

Lumbar surgical operation is the crucial treatment against lumbar degenerative diseases (LDDs), whose development depends on persistent comprehension and innovation of vertebral biomechanics. The thorough understanding of biomechanical changes during lumbar senescence and degeneration is the important bedrock to grasp LDDs pathogenesis, renovate LDDs surgical strategy, and embrace more precise and minimally invasive treatment against LDDs. Herein, in this review, the intimate crosstalk between LDDs with degenerative biomechanics of vertebrae, intervertebral disc and paravertebral muscles was elucidated, followed by the classification of lumbar surgery history into non-vertebral implant era (before the year 1980), vertebral implant era (during the year 1980-1990), vertebral fusion era (during the year 1990-2010), precise and minimally invasive decompression era (after the year 2010) based on lumbar surgical characteristics in each era. The significance of representative biomechanical studies in each era for lumbar surgery was also concluded. From biomechanical perspectives, the history of spinal surgery is the development history of surgical strategies that has progressed as the continuously in-depth understanding of spinal biomechanics. With the deepening of spinal biomechanical researches, spinal surgeons are expected to develop treatment strategies that are more adapted to physiological and biomechanical characteristics of the spine, thereby guiding the future direction of spinal surgery advancement.

3.
Acta Pharmaceutica Sinica B ; (6): 735-746, 2022.
Article in English | WPRIM | ID: wpr-929323

ABSTRACT

The cell cycle inhibitor P21 has been implicated in cell senescence and plays an important role in the injury-repair process following lung injury. Pulmonary fibrosis (PF) is a fibrotic lung disorder characterized by cell senescence in lung alveolar epithelial cells. In this study, we report that P21 expression was increased in alveolar epithelial type 2 cells (AEC2s) in a time-dependent manner following multiple bleomycin-induced PF. Repeated injury of AEC2s resulted in telomere shortening and triggered P21-dependent cell senescence. AEC2s with elevated expression of P21 lost their self-renewal and differentiation abilities. In particular, elevated P21 not only induced cell cycle arrest in AEC2s but also bound to P300 and β-catenin and inhibited AEC2 differentiation by disturbing the P300-β-catenin interaction. Meanwhile, senescent AEC2s triggered myofibroblast activation by releasing profibrotic cytokines. Knockdown of P21 restored AEC2-mediated lung alveolar regeneration in mice with chronic PF. The results of our study reveal a mechanism of P21-mediated lung regeneration failure during PF development, which suggests a potential strategy for the treatment of fibrotic lung diseases.

4.
Acta Pharmaceutica Sinica B ; (6): 665-677, 2022.
Article in English | WPRIM | ID: wpr-929318

ABSTRACT

Sulfonylureas are widely used oral anti-diabetic drugs. However, its long-term usage effects on patients' lifespan remain controversial, with no reports of influence on animal longevity. Hence, the anti-aging effects of chlorpropamide along with glimepiride, glibenclamide, and tolbutamide were studied with special emphasis on the interaction of chlorpropamide with mitochondrial ATP-sensitive K+ (mitoK-ATP) channels and mitochondrial complex II. Chlorpropamide delayed aging in Caenorhabditis elegans, human lung fibroblast MRC-5 cells and reduced doxorubicin-induced senescence in both MRC-5 cells and mice. In addition, the mitochondrial membrane potential and ATP levels were significantly increased in chlorpropamide-treated worms, which is consistent with the function of its reported targets, mitoK-ATP channels. Increased levels of mitochondrial reactive oxygen species (mtROS) were observed in chlorpropamide-treated worms. Moreover, the lifespan extension by chlorpropamide required complex II and increased mtROS levels, indicating that chlorpropamide acts on complex II directly or indirectly via mitoK-ATP to increase the production of mtROS as a pro-longevity signal. This study provides mechanistic insight into the anti-aging effects of sulfonylureas in C. elegans.

5.
Acta Pharmaceutica Sinica B ; (6): 511-531, 2022.
Article in English | WPRIM | ID: wpr-929312

ABSTRACT

Aging is by far the most prominent risk factor for Alzheimer's disease (AD), and both aging and AD are associated with apparent metabolic alterations. As developing effective therapeutic interventions to treat AD is clearly in urgent need, the impact of modulating whole-body and intracellular metabolism in preclinical models and in human patients, on disease pathogenesis, have been explored. There is also an increasing awareness of differential risk and potential targeting strategies related to biological sex, microbiome, and circadian regulation. As a major part of intracellular metabolism, mitochondrial bioenergetics, mitochondrial quality-control mechanisms, and mitochondria-linked inflammatory responses have been considered for AD therapeutic interventions. This review summarizes and highlights these efforts.

6.
Article in English | WPRIM | ID: wpr-929018

ABSTRACT

OBJECTIVES@#Liver cancer is the sixth most common malignant tumor in the world. Hepatocellular carcinoma (HCC) accounts for 85%-90% of all patients with liver cancer. It possesses the characteristics of insidious onset, rapid progression, early recurrence, easy drug resistance, and poor prognosis. NIMA related kinase 2 (NEK2) is a cell cycle regulating kinases, which regulates cell cycle in mitosis. Cellular senescence is a complex heterogeneous process, and is a stable form of cell cycle arrest that limits the proliferative potential of cells. This study aims to investigate the relationship between the expression level of NEK2 and the senescence in hepatoma cells, and to explore the effect of NEK2 expression on hepatoma cell senescence and the underlying molecular mechanism.@*METHODS@#A total of 581 senescence-relevant genes were obtained from the GenAge website. The gene expression data of tumor tissues of 370 HCC patients were downloaded from the Cancer Genome Atlas database. The co-expression of NEK2 and aging-related genes was analyzed by R-package. KEGG was used to analyze the significant gene enrichment pathway of differentially expressed genes in NEK2 overexpression HEK293. The stable transfected cell lines with overexpression and knockdown of NEK2 were constructed in hepatoma cell line SMMC-7721 and HepG2, and senescence-associated β-galactosidase (SA-β-gal) staining was used to detect senescence, the cell proliferation was detected by CCK-8 method and clone formation experiment, the cell cycle was analyzed by flow cytometry, and the expression of proteins related to p53/p21, p16/Rb, and phosphatase and tensin homolog deleted on chromosome ten (PTEN)/Akt signal transduction pathway was detected by Western blotting.@*RESULTS@#There were 320 senescence related genes co-expressed with NEK2. KEGG analysis showed that the senescence signaling pathway was significantly enriched in HEK293 cells with overexpression of NEK2.Compared with SMMC-7721 or HepG2 without knockdown of NEK2, the senescent cells of SMMC-7721 and HepG2 with knockdown of NEK2 were increased, cell proliferation and clone formation were decreased significantly, the percentage of cells in G0/G1 phase was increased, the expression levels of phospho-Akt (p-Akt) and phospho-Rb (p-Rb) protein were decreased significantly, and the expression level of p16 protein was increased significantly (all P<0.05). Compared with SMMC-7721 or HepG2 transfected with blank plasmid, the senescent cells of SMMC-7721 and HepG2 overexpressing NEK2 were decreased, the cell proliferation and clone formation were increased significantly, the percentage of cells in G0/G1 phase were decreased, the expression levels of p-Akt and p-Rb protein were increased significantly, and the expression level of p16 protein was decreased significantly (all P<0.05).@*CONCLUSIONS@#NEK2 may mediate the anti-aging effect of hepatoma cells through p16/Rb and PTEN/Akt signal transduction pathways, which provides a new theoretical basis for NEK2 to promote the progress of liver cancer and a new idea for the targeting treatment for liver cancer.


Subject(s)
Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Cell Proliferation/physiology , Cellular Senescence/genetics , HEK293 Cells , Humans , Liver Neoplasms/pathology , NIMA-Related Kinases/genetics , Proto-Oncogene Proteins c-akt/metabolism
7.
Article in English | WPRIM | ID: wpr-928660

ABSTRACT

Cellular senescence is a biological process associated with the degeneration of cell structure and function, which contribute to age-related diseases. Atherosclerosis is a chronic inflammatory disease that can cause a variety of cardiovascular disorders. In this article, we review the effects of cellular senescence on the development of atherosclerosis through diverse physiopathological changes, focusing on the alterations in senescent organelles and the increased senescence-associated secretory phenotype (SASP), and exploring the relevant therapeutic strategies for atherosclerosis by clearing senescent cells and reducing SASP, to provide new insights for the treatment of atherosclerosis.


Subject(s)
Aging , Atherosclerosis , Cardiovascular Diseases , Cellular Senescence , Chronic Disease , Humans , Senescence-Associated Secretory Phenotype
8.
Arq. bras. med. vet. zootec. (Online) ; 73(6): 1413-1421, Nov.-Dec. 2021. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1355684

ABSTRACT

The objective this study was to identify differences in the morphogenic patterns of four Urochloa brizantha cultivars (marandu, piatã, xaraés and paiaguás grasses) during the stockpiling period. A completely randomized design was used, with three replications, in experimental plots of 9m². The evaluations took place over 2 years. The grasses were stockpiled for 92 (Year 1) and 95 (Year 2) days. The leaf appearance rate of paiaguás grass was higher, compared to other grasses. In Year 1, the stem elongation rate of xaraés grass was higher than other grasses. At the end of stockpiling period of Year 1, the tiller population density (TPD) was higher in the paiaguás grass, intermediate in the xaraés and marandu grasses and lower in the piatã grass. At the end of the stockpiling period in Year 2, TPD was higher in the paiaguás grass canopy, intermediate in the marandu and piatã grasses canopies, and lower in the xaraés grass canopy. Paiaguás grass has greater leaf growth during the stockpiling period and is therefore suitable for use under stockpiled grazing. Xaraés grass has high stem elongation during the stockpiling period, which is why its use under stockpiled grazing must be accompanied by adjustments in pasture management.(AU)


O objetivo deste estudo foi identificar diferenças nos padrões morfogênicos de quatro cultivares de Urochloa brizantha (marandu, piatã, xaraés e paiaguás) durante o período de diferimento. Foi utilizado o delineamento inteiramente ao acaso, com três repetições, em parcelas experimentais de 9m². As avaliações ocorreram por dois anos. Os capins foram diferidos por 92 (Ano 1) e 95 (Ano 2) dias. A taxa de aparecimento foliar do capim-paiaguás foi maior, em comparação aos demais capins. No Ano 1, a taxa de alongamento do colmo do capim-xaraés foi superior aos demais capins. No final do diferimento do Ano 1, a densidade populacional de perfilho (DPP) foi maior no capim-paiaguás, intermediária nos capins xaraés e marandu e inferior no capim-piatã. No fim do período de diferimento do Ano 2, a DPP foi superior no dossel de capim-paiaguás, intermediária nos dosséis dos capins marandu e piatã, e menor no dossel de capim-xaraés. O capim-paiaguás tem maior crescimento foliar durante o período de diferimento, sendo, portanto, apropriado para uso sob pastejo diferido. O capim-xaraés apresenta elevado alongamento de colmo durante o período de diferimento, razão pela qual seu uso sob pastejo diferido deve vir acompanhado de ajustes no manejo da pastagem.(AU)


Subject(s)
Plant Stems , Brachiaria/growth & development , Morphogenesis/physiology
9.
Rev. Assoc. Med. Bras. (1992) ; 67(2): 173-177, Feb. 2021.
Article in English | LILACS | ID: biblio-1287831

ABSTRACT

SUMMARY The aging process occurs due to the decline of vital physiological functions and adaptability of the body, being influenced by genetics and lifestyle. With advances in genetics, biological aging can be calculated by telomere length. Telomeres are regions at the ends of chromosomes that play a role in the maintenance and integrity of DNA. With biological aging, telomere shortening occurs, causing cellular senescence. Several studies show that shorter telomeres are associated with acute and chronic diseases, stress, addictions, and intoxications. Even in the current COVID-19 pandemic, telomere shortening is proposed as a marker of severity in individuals infected by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). On the other hand, healthy lifestyle habits increase telomere length and balance of various cellular functions, preventing diseases.


Subject(s)
Humans , COVID-19 , Aging/genetics , Biomarkers , Telomere/genetics , Pandemics , SARS-CoV-2
10.
Article in Chinese | WPRIM | ID: wpr-921779

ABSTRACT

This study aims to explore the effect of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma(hereinafter referred to as GNS) on the SIRT1-autophagy pathway of endothelial cell senescence induced by hydrogen peroxide(H_2O_2). To be specific, vascular endothelial cells were classified into the blank control group(control), model group(model), model + DMSO group(DMSO), resveratrol group(RESV), and GNS low-dose(GNS-L), medium-dose(GNS-M), and high-dose(GNS-H) groups. They were treated with H_2O_2 for senescence induction except the control. After intervention of cells in each group with corresponding drugs for 24 h, cell growth status was observed under an inverted microscope, and the formation of autophagosome under the transmission electron microscope. In addition, the changes of microtubule-associated protein 1 light chain 3β(LC3 B) were detected by immunofluorescence staining. The autophagy flux was tracked with the autophagy double-labeled adenovirus(mRFP-GFP-LC3) fusion protein. Dansylcadaverine(MDC) staining was employed to determine the autophagic vesicles, and Western blot the expression of sirtuin 1(SIRT1), ubiquitin-binding protein p62, and LC3Ⅱ. After H_2O_2 induction, cells demonstrated slow growth, decreased adhesion ability, raised number of SA-β-gal-stained blue ones, a certain number of autophagosomes with bilayer membrane and secondary lysosomes in the cytoplasm, and slight rise of autophagy flux level. Compared with the model group, GNS groups showed improved morphology, moderate adhesion ability, complete and smooth membrane, decreased SA-β-gal-stained blue cells, many autophagosomes, autophagic vesicles, and secondary lysosomes in the cytoplasm, increased autophagolysosomes, autophagy flux level, and fluorescence intensity of LC3 B and MDC, up-regulated expression of SIRT1 and LC3Ⅱ, and down-regulated expression of p62, suggesting the improvement of autophagy level. GNS can delay the senescence of vascular endothelial cells. After the intervention, the autophagy flux and related proteins SIRT1, LC3Ⅱand p62 changed significantly, and the autophagy level increased significantly. However, EX527 weakened the effect of Chinese medicine in delaying vascular senescence. GNS may delay the senescence of vascular endothelial cells through the SIRT1 autophagy pathway.


Subject(s)
Autophagy , Cells, Cultured , Cellular Senescence , Drugs, Chinese Herbal/pharmacology , Endothelial Cells/drug effects , Hydrogen Peroxide , Panax/chemistry , Sirtuin 1/genetics
11.
Article in Chinese | WPRIM | ID: wpr-912501

ABSTRACT

Objective:To explore the age-related changes of Pole2 expression levels in peripheral blood lymphocytes of healthy people, and analyze the differences of Pole2 expression levels in peripheral blood lymphocytes of four common senile disease patients between normal peers.Methods:Healthy people and patients in the physical examination center of Xuanwu Hospital of Capital Medical University from December 2018 to December 2019 were selected as the research objects, the mRNA and protein level of Pole2 in heathy individual as 20-29,30-39,40-49,50-59,60-69,70-79 y were checked by RT-PCR and Western blot,and the age-related curve was drew. The mRNA and protein expression levels of Pole2 were also detected in the peripheral blood lymphocytes of patients of DM,CHD,AD,CVD. And the deviation with healthy people of the same age was analyzed from one way ANOVA and repeated measurement were used to compare the mean of multiple groups.Results:The mRNA and protein levels of Pole2 increased in the lymphocytes of healthy people at 20-29 y and 50-59 y grouy, and decreased after 60-69 y and 70-79 y ( P<0.001). The Pole2 levels of mRNA and protein in the lymphocytes of patients of CHD,AD,DM and CVD cerebral atherosclerosis were significantly higher than that of healthy people of the same age. Moreover, the Pole2 level of lymphocytes of AD patients was significantly higher than that of other patient groups ( P<0.001). Conclusion:The expression level of Pole2 in peripheral blood lymphocytes increases with age before the age of 60 years old and decreases with age after 60 years old ;and the expression in four common senile diseases was higher than that in normal people.

12.
Article in Chinese | WPRIM | ID: wpr-910417

ABSTRACT

Objective:To explore the mechanism and regulatory effects of melatonin on UVB-induced melanin synthesis in human immortalized keratinocytes (HaCaT), so as to provide a theoretical basis for the skin protection of melatonin.Methods:HaCaT cells were pretreated with 10 -5 mol/L melatonin and then irradiated with 80 mJ/cm 2UVB. The melanin content was detected by NaOH assay, the proportion of premature senescence cells was detected by β-galactosidase staining kit, and the protein expression levels of both p53 and tyrosinase (TYR) were detected by Western blot at 72 h after UVB exposure. After 12 h pretreatment of ATM/ATR inhibitor, p53 inhibitor and melatonin, the proportion of premature senescence and the change of melanin content in HaCaT cells were detected at 72 h after 80 mJ/cm 2 UVB irradiation. Results:Melatonin inhibited UVB-induced increases of melanin content ( t=56.65, 13.39, P<0.05) and TYR expression ( t=16.46, P<0.05) in HaCaT cells. Melatonin alleviated UVB-induced premature senescence ( t=7.139, P<0.05) and inhibited UVB-induced increase of p53 expression ( t=19.08, P<0.05) in HaCaT cells. In addition, ATM/ATR inhibitor, p53 inhibitor and melatonin all inhibited UVB-induced increase of melanin content in HaCaT cells. Conclusions:Melatonin inhibits TYR-mediated melanin synthesis by regulating p53-related premature senescence in HaCaT cells after UVB irradiation.

13.
Article in Chinese | WPRIM | ID: wpr-906394

ABSTRACT

Objective:To investigate the effects of Guiqi Dingnian prescription (GDP) on the expression of related molecules in Janus tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription (JAK2/STAT) signaling pathway of <italic>D</italic>-galactose (<italic>D</italic>-gal)-induced senescent mesangial cells. Method:The senescent mouse mesangial cells induced by 10 g·L<sup>-1</sup> <italic>D</italic>-gal were continuously treated with 40 mg·L<sup>-1 </sup>GDP for three days. The senescence of the treated cells was determined by senescence-associated (SA)-<italic>β</italic>-gal staining. The cell cycle was detected by flow cytometry. The cell viability was analyzed using the cell counting kit-8 (CCK-8). The mRNA expression levels of tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>), interleukin-6 (IL-6), nuclear transcription factor-<italic>κ</italic>B (NF-<italic>κ</italic>B), and IL-1<italic>α</italic> were detected by real-time polymerase chain reaction (Real-time PCR). The protein expression levels of STAT1, phosphorylated STAT1 (p-STAT1), STAT3, and p-STAT3 in the JAK2/STAT signaling pathway were determined by Western blot. Result:CCK-8 results showed that the optimal concentration of GDP was 40 mg·L<sup>-1</sup>. Compared with the blank group, the positive rate of SA-<italic>β</italic>-gal in the model group was significantly higher(<italic>P</italic><0.01), the percentage of cells in G<sub>0</sub>/G<sub>1</sub> phase was significantly increased(<italic>P</italic><0.05), the percentage of cells in G<sub>2</sub>/M and S phase was significantly decreased(<italic>P</italic><0.01). The mRNA expression levels of TNF-<italic>α</italic>,IL-6,NF-<italic>κ</italic>B and IL-1<italic>α </italic>were significantly increased(<italic>P</italic><0.01). Compared with the model group, the model + GDP group exhibited significantly decreased SA-<italic>β</italic>-gal-positive cells (<italic>P</italic><0.05), reduced cells in the G<sub>0</sub>/G<sub>1</sub> phase (<italic>P</italic><0.05), increased cells in the G<sub>2</sub>/M and S phases (<italic>P</italic><0.01), and down-regulated TNF-<italic>α</italic>, IL-6, NF-<italic>κ</italic>B, and IL-1<italic>α </italic>mRNA expression (<italic>P</italic><0.05) and STAT1, p-STAT1, STAT3, and p-STAT3 protein expression (<italic>P</italic><0.05). Conclusion:GDP delays the senescence of mouse mesangial cells possibly by down-regulating the expression of related molecules in the JAK2/STAT pathway.

14.
Article in Chinese | WPRIM | ID: wpr-906174

ABSTRACT

Objective:To investigate the effect of Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract on endothelial microparticles (EMPs)-induced vascular endothelial cell senescence, and explore the possible mechanism. Method:Human umbilical vein endothelial cells (HUVECs) were used as the research objects, and the aged model was established with 10-12 passages of replicative senescence cells. The experimental cells were divided into young group (2-4 passage cells), aged group (10-12 passage cells), only EMPs intervention group (extract EMPs produced by aged cells to intervene young cells) and low dose, middle dose and high dose drug intervention groups (200, 300, 400 mg·L<sup>-1</sup>). Senescence related <italic>β</italic>-galactosidase (SA-<italic>β</italic>-gal) staining and cell cycle propidium iodide (PI) staining were used to determine cell senescence. Cell counting kit-8 (CCK-8) assay was used to screen the drug concentration. EMPs were extracted by two-step centrifugation, EMPs labeled with phycoerythrin (PE) anti-human CD31 antibody or fluorescein isothiocyanate (FITC) annexin V were detected by flow cytometry, intracellular reactive oxygen species (ROS) were detected by 2',7'- dichlorofluorescein diacetate (DCFDA) staining. Result:After treatment with the drug, SA-<italic>β</italic>-gal activity of the aged cells significantly decreased (<italic>P</italic><0.01), the S phase arrest was restored (<italic>P</italic><0.01), and the number of CD31<sup>+</sup> EMPs and annexin V<sup>+</sup> EMPs secreted by aged cells decreased (<italic>P</italic><0.05). Compared with the young group, only EMPs intervention group could induce increased SA-<italic>β</italic>-gal activity and S phase arrest in young cells (<italic>P</italic><0.05,<italic>P</italic><0.01). However, after intervention of EMPs and the drug, EMPs-mediated increase of SA-<italic>β</italic>-gal activity was significantly inhibited and S phase arrest was restored (<italic>P</italic><0.05). The increase of intracellular ROS induced by EMPs was also significantly inhibited by the drug (<italic>P</italic><0.05,<italic>P</italic><0.01). Conclusion:Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract can delay the senescence of vascular endothelial cells by influencing EMPs, and the mechanism may be related to the inhibition of increased intracellular ROS induced by EMPs.

15.
Acta Pharmaceutica Sinica B ; (6): 3935-3949, 2021.
Article in English | WPRIM | ID: wpr-922451

ABSTRACT

A significant proportion of non-small cell lung cancer (NSCLC) patients experience accumulating chemotherapy-related adverse events, motivating the design of chemosensitizating strategies. The main cytotoxic damage induced by chemotherapeutic agents is DNA double-strand breaks (DSB). It is thus conceivable that DNA-dependent protein kinase (DNA-PK) inhibitors which attenuate DNA repair would enhance the anti-tumor effect of chemotherapy. The present study aims to systematically evaluate the efficacy and safety of a novel DNA-PK inhibitor M3814 in synergy with chemotherapies on NSCLC. We identified increased expression of DNA-PK in human NSCLC tissues which was associated with poor prognosis. M3814 potentiated the anti-tumor effect of paclitaxel and etoposide in A549, H460 and H1703 NSCLC cell lines. In the four combinations based on two NSCLC xenograft models and two chemotherapy, we also observed tumor regression at tolerated doses

16.
Acta Pharmaceutica Sinica ; (12): 1137-1146, 2021.
Article in Chinese | WPRIM | ID: wpr-886996

ABSTRACT

Using a H2O2-induced BRL cell senescence model, we investigated the anti-aging effects of drug-containing serums of Erzhi Wan (EZW) and various polar extracts (petroleum ether, ethyl acetate, n-butanol, water, and iridoid glycoside-enriched fractions). Cell viability was detected by MTT assay. Cell senescence was evaluated with β-galactosidase staining assay. Intracellular reactive oxygen species (ROS) were measured by flow cytometry. UFLC-Q-TOF-MS/MS was used to identify chemical components in EZW and the extracts, and molecular docking technology was used to predict the anti-aging components of EZW. Results showed that treatment of cells with 600 μmol·L-1 H2O2 for 72 h markedly induced cell senescence, inhibited cell proliferation and increased intracellular β-galactosidase activity and ROS levels. If cells were pretreated with drug-containing serum of EZW this induction of senescence was decreased. A total of 49 chemical compounds were identified in EZW by liquid chromatography-mass spectrometry, 14 of these were identified by molecular docking as potential active ingredients. Based on these analyses, and the in vitro experiments with polar extracts, we conclude that the anti-aging components of EZW are triterpenes, flavonoids and phenyl alcohols, providing a basis for further elucidation of the active agents and mechanism of the anti-aging effect of EZW.

17.
Article in Chinese | WPRIM | ID: wpr-883378

ABSTRACT

Objective:To determine the effect of rice bran extract (RBE) in combination with doxorubicin on 4T1 triple-negative breast cancer cells and NIH-3T3 cells. Methods:RBE was obtained by maceration with n-hexane. The phytochemical profile of RBE was observed using high-performance liquid chromatography. Cytotoxic effect of RBE was evaluated through MTT assay. In addition, flow cytometry was used for cell cycle and apoptosis analysis. Cellular senescence was observed using SA-β-Gal assay and intracellular reactive oxygen species (ROS) levels were evaluated using DCFDA staining. The pro-oxidant property of RBE was also evaluated through 1-chloro-2,4-dinitrobenzene spectrophotometry and molecular docking. Results:RBE was obtained with a yield of 18.42% w/w and contained tocotrienols as the major compound. RBE exerted no cytotoxic effect on 4T1 and NIH-3T3 cells. However, RBE in combination with doxorubicin decreased 4T1 cell viability synergistically (combination index<0.9) and induced apoptosis and senescence on 4T1 cells. RBE significantly decreased senescence in doxorubicin-treated NIH-3T3 cells. Additionally, RBE did not increase ROS levels in doxorubicin-treated 4T1 cells. Meanwhile, the combination of RBE and doxorubicin reduced ROS levels in NIH-3T3 cells. RBE significantly reduced glutathione-S-transferase activity and alpha-tocotrienol interacted with glutathione-S-transferase in the glutathione binding site. Conclusions:Rice bran may be used as a co-chemotherapeutic agent to improve the therapeutic effectiveness of doxorubicin while protecting against the cellular senescence effects of doxorubicin on healthy cells.

18.
Article in Chinese | WPRIM | ID: wpr-883368

ABSTRACT

Objective:To investigate the potential anti-aging mechanism of 9-hydroxy-6,7-dimethoxydalbergiquinol (HDDQ) on hydrogen peroxide (H2O2)-induced oxidative stress in human dermal fibroblasts (HDFs). Methods:The effect of HDDQ on cell viability was assessed by MTT assay, and the effects of HDDQ on senescence-like phenotypes were determined by senescence-associated β-galactosidase (SA-β-gal) staining, Western blotting analysis, and a cell proliferation assay. The expression level and activity of sirtuin-1 (SIRT1) induced by HDDQ were also measured. Results:HDDQ reversed senescence-like phenotypes in the oxidant-challenged model, through reducing SA-β-gal activity and promoting cell growth. Meanwhile, decreases in ac-p53, p21Cip1/WAF1, and p16Ink4a and an increase in pRb were observed. HDDQ induced the expression of SIRT1 in a concentration- and time-dependent manner. Moreover, HDDQ inhibited H2O2-induced phosphorylation of Akt by SIRT1 up-regulation and reduced SA-β-gal staining. Conclusions:HDDQ inhibits H2O2-induced premature senescence and upregulation of SIRT1 expression plays a vital role in the inhibition of the senescence phenotype in HDFs.

19.
Article in Chinese | WPRIM | ID: wpr-883366

ABSTRACT

Objective:To investigate the anti-senescence effect of 3-bromo-4,5-dihydroxybenzaldehyde (BDB) from Polysiphonia morrowii Harvey in human dermal fibroblasts (HDF). Methods:HDF were subjected to treatment of BDB and then treated with hydrogen peroxide (H2O2) to induce premature senescence. Senescence-associated β-galactosidase (SA-β-gal) activity in HDF was determined using the SA-β-gal staining method. Intracellular reactive oxygen species (ROS) production was measured using the 2',7'-dichlorodihydrofluorescein diacetate assay. Western blotting assay was performed to assess the level of antioxidant enzyme glutathione peroxidase 1 (GPX1). In addition, intracellular collagen and collagenase contents were analyzed using the respective ELISA kits. Elastase activity in HDF supernatants was measured from p-nitroaniline release and normalized using total protein content. Results:Treatment of HDF with H2O2 increased the activity of SA-β-gal, but BDB pre-treatment resulted in the reduction of SA-β-gal activity. Moreover, BDB significantly reduced H2O2-induced intracellular ROS production. BDB also markedly increased the level of GPX1, which was inhibited by 400 μM of H2O2. Furthermore, in in vitro study, BDB significantly increased intracellular collagen content and decreased matrix metalloproteinase-1 and elastase activities in HDF. Conclusions:Our results demonstrate that BDB shows anti-senescence and anti-wrinkle activities in vitro.

20.
Acta Pharmaceutica Sinica B ; (6): 340-354, 2021.
Article in English | WPRIM | ID: wpr-881140

ABSTRACT

Enormous studies have corroborated that long non-coding RNAs (lncRNAs) extensively participate in crucial physiological processes such as metabolism and immunity, and are closely related to the occurrence and development of tumors, cardiovascular diseases, nervous system disorders, nephropathy, and other diseases. The application of lncRNAs as biomarkers or intervention targets can provide new insights into the diagnosis and treatment of diseases. This paper has focused on the emerging research into lncRNAs as pharmacological targets and has reviewed the transition of lncRNAs from the role of disease coding to acting as drug candidates, including the current status and progress in preclinical research. Cutting-edge strategies for lncRNA modulation have been summarized, including the sources of lncRNA-related drugs, such as genetic technology and small-molecule compounds, and related delivery methods. The current progress of clinical trials of lncRNA-targeting drugs is also discussed. This information will form a latest updated reference for research and development of lncRNA-based drugs.

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