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Understanding species boundaries maintenance in the face of hybridization/introgression is an intriguing yet complex topic in evolutionary biology. The underlying mechanisms, however, remain elusive. To address this, we propose to investigate the role of climatic shifts in shaping genetic structure and influencing species boundaries. We combine multilocus genetic data and species distribution modeling to explore how past and current climatic shifts affect the genetic structure and demographic history of two Taiwan endemic gingers, Zingiber pleiostachyum and Z. shuanglongense. We identified a well-delimited genetic structure with evidence of admixture, indicating incomplete reproductive isolation between the two gingers. This is likely due to secondary contact and range overlap during the last glacial maximum, leading to sporadic instances of hybridization. Niche overlap tests based on climate and soil data indicate that these two gingers occupy similar but nonidentical ecological niches. Furthermore, we found that the considerable differences in their current geographic distribution and altitude preferences might have resulted from different seed dispersal capabilities and competitive exclusion due to their similar niche preferences. Our results reveal a model where altitudinal differentiation and dispersal strategy synergistically reinforce the species divergence, thereby illuminating the importance of these factors in shaping and maintaining the island's biodiversity.
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Zingiber officinale , Ecossistema , Evolução Biológica , Hibridização Genética , Hibridização de Ácido Nucleico , FilogeniaRESUMO
Osteoporosis is a generalized disease of bone that leads to a loss of bone density and bone mass, destruction of bone microstructure, increased brittleness and therefore fracture. At present, the main treatment of Western medicine is drug therapy such as bisphosphonates, calcitriol, vitamin D, etc. However, long-term use of these drugs may bring some adverse reactions. Chinese herbal medicine Cistanche deserticola could regulate bone metabolism by promoting osteoblast activity and inhibiting osteoclast activity with low toxicity and adverse reactions. Therefore, Cistanche deserticola has attracted increasing attention for its efficacy in the prevention and treatment of osteoporosis in recent years. Here we present a literature review of the molecular pathways involved in osteoporosis and the effects of Cistanche deserticola on bone metabolism. Our objective is to clarify the mechanism of Cistanche deserticola in the treatment of osteoporosis.
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Cistanche , Osteoporose Pós-Menopausa , Osteoporose , Feminino , Humanos , Osteoporose Pós-Menopausa/tratamento farmacológico , Cistanche/química , Osteoclastos , Osteoporose/tratamento farmacológico , Densidade ÓsseaRESUMO
Brackish water irrigation increases soil salinity and changes the soil environment, which affects the structure and diversity of soil fungi. In this study, the effects of biochar and straw (3.7 t·hm-2 and 6 t·hm-2, respectively) on soil physical and chemical properties and fungal community structure diversity were investigated on the basis of long-term brackish water irrigation. The results showed that compared to the absence of biochar and straw application (control), biochar application significantly increased pH and the contents of total carbon, available potassium, and available phosphorus in soil but significantly decreased the soil conductivity by 20.71%. Straw treatment significantly increased the content of available potassium and phosphorus but significantly decreased the soil bulk density and conductivity by 4.17% and 64.50%, respectively. The biochar and straw treatment showed an increasing trend in the Chao1 index and ACE index of the fungal community but a decreasing trend in the Shannon index and Simpson index. The dominant fungal phyla in the soil were Ascomycota, Mortierellomycota, Basidiomycota, Chytridiomycota, and Glomeromycota. The dominant fungal genera were Chaetomium, Gibberella, Fusarium, Idriella, and Mortierella. Biochar and straw were applied to increase the relative abundance of Ascomycota, Mortierellomycota, Basidiomycota, Glomeromycota, and Chaetomium. However, the relative abundance of Chytridomycota, Gibberella, and Idriella decreased. LEfSe analysis showed that biochar application and straw returning decreased the number of potential biomarkers in fungal communities. RDA results showed that soil fungal community structure was significantly correlated with EC1:5 and TN. Brackish irrigation had adverse effects on soil, in which EC1:5and TN were the main factors driving the change in soil fungal community structure. The soil fungal community adapted to a salt-stress environment through the improvement of soil by biochar and straw.
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Micobioma , Carvão Vegetal , Fósforo , Potássio , Águas Salinas , Solo/química , Microbiologia do SoloRESUMO
Long-term saline water irrigation will increase soil salinity, adversely affect soil physical and chemical properties, and change the diversity of soil bacteria. Straw returning can improve the soil microenvironment and subsequently affect soil enzyme activity and bacterial community structure diversity. This experiment used two types of irrigation water salinity:fresh water (FW, 0.35 dS·m-1) and saline water (SW, 8.04 dS·m-1). Under each irrigation water salinity, the amount of cotton straw applied was 0 and 6000 kg·hm-2 (represented by FWST and SWST, respectively). The results showed that:compared with those under fresh water irrigation, saline water irrigation significantly increased the soil salt, bulk density, total carbon, and available phosphorus but significantly decreased available potassium content. Under saline water irrigation, straw returning significantly increased the soil total carbon, total nitrogen, available potassium, and available phosphorus but reduced soil bulk density. Compared with those under fresh water irrigation, soil sucrase, alkaline phosphatase, and catalase activities under saline water irrigation decreased by 57.24%, 35.15%, and 3.91%, respectively, whereas urease activity increased by 26.73%. However, straw returning significantly increased sucrase, alkaline phosphatase, and catalase activities but decreased urease activity. Saline water irrigation decreased the relative abundance of Acidobacteriota, Actinobacteriota, Bacteroidota, Verrucomicrobiota, and Firmicutes and increased the abundance of Gemmatimonadota and Myxococcota. Under saline water irrigation, straw returning significantly increased the relative abundance of Actinobacteriota, Bacteroidetes, Firmicutes, Crenarchaeota, Sphingomonas, Dongia, and Steroidobacter. NMDS results also showed that saline water irrigation and straw returning changed the bacterial community structure. In conclusion, straw returning can improve soil nutrient content, reduce soil bulk density and salinity, and then change soil enzyme activity and bacterial community structure diversity. The change in soil bacterial community composition was mainly affected by soil salinity and bulk density. Therefore, straw returning can improve soil fertility and help maintain the health of soil ecosystem. This study revealed a relationship between soil enzyme activities and bacterial communities, which provides a theoretical basis and mechanism for applying cotton stalk to regulate the soil enzyme and micro-ecological environment.
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Ecossistema , Solo , Fosfatase Alcalina , Bactérias , Carbono , Catalase , Fósforo , Potássio , Águas Salinas , Solo/química , Microbiologia do Solo , Sacarase , UreaseRESUMO
Loganin is an iridoid glycoside extracted from Cornus officinalis, which is a traditional oriental medicine, and many biological properties of loganin have been reported. Nevertheless, it is not clear whether loganin has therapeutic effect on cardiovascular diseases. Hence, the aim of the present study was to investigate the effect of loganin on Ang II-induced cardiac hypertrophy. In the present study, we reported for the first time that loganin inhibits Ang II-provoked cardiac hypertrophy and cardiac damages in H9C2 cells and in mice. Furthermore, loganin can achieve cardioprotective effects through attenuating cardiac fibrosis, decreasing pro-inflammatory cytokine secretion, and suppressing the phosphorylation of critical proteins such as JAK2, STAT3, p65, and IκBα. Besides, the outstanding findings of the present study were to prove that loganin has no significant toxicity or side effects on normal cells and organs. Based on these results, we conclude that loganin mitigates Ang II-induced cardiac hypertrophy at least partially through inhibiting the JAK2/STAT3 and NF-κB signaling pathways. Accordingly, the natural product, loganin, might be a novel effective agent for the treatment of cardiac hypertrophy and heart failure.
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Multiplexed optical imaging provides holistic visualization on a vast number of molecular targets, which has become increasingly essential for understanding complex biological processes and interactions. Vibrational microscopy has great potential owing to the sharp linewidth of vibrational spectra. In 2017, we demonstrated the coupling between electronic pre-resonant stimulated Raman scattering (epr-SRS) microscopy with a proposed library of 9-cyanopyronin-based dyes, named Manhattan Raman Scattering (MARS). Herein, we develop robust synthetic methodology to build MARS probes with different core atoms, expansion ring numbers, and stable isotope substitutions. We discover a predictive model to correlate their vibrational frequencies with structures, which guides rational design of MARS dyes with desirable Raman shifts. An expanded library of MARS probes with diverse functionalities is constructed. When coupled with epr-SRS microscopy, these MARS probes allow us to demonstrate not only many versatile labeling modalities but also increased multiplexing capacity. Hence, this work opens up next-generation vibrational imaging with greater utilities.
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Corantes/química , Sondas Moleculares/química , Microscopia Óptica não Linear/métodos , Imagem Óptica/métodos , Pironina/química , Corantes/síntese química , Células HeLa , Humanos , Modelos Químicos , Sondas Moleculares/síntese química , Estrutura Molecular , Pironina/análogos & derivados , Pironina/síntese química , Análise Espectral Raman/métodos , VibraçãoRESUMO
Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers, which lacks effective treatment strategies. There is an urgent need for the development of new strategies for PDAC therapy. The genetic and phenotypic heterogeneity of PDAC cancer cell populations poses further challenges in the clinical management of PDAC. In this study, we performed single-cell RNA sequencing to characterize PDAC tumors from KPC mice. Functional studies and clinical analysis showed that PDAC cluster 2 cells with highly Hsp90 expression is much more aggressive than the other clusters. Genetic and pharmacologic inhibition of Hsp90 impaired tumor cell growth both in vitro and in vivo. Further mechanistic study revealed that HSP90 inhibition disrupted the interaction between HSP90 and OPA1, leading to a reduction in mitochondrial cristae amount and mitochondrial energy production. Collectively, our study reveals that HSP90 might be a potential therapeutic target for PDAC.
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Objective:An ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was developed to analyze and identify the chemical constituents in <italic>Coptis chinensis</italic> inflorescence. Method:The chromatographic separation was performed on ACQUITY UPLC BEH C<sub>18</sub> column (2.1 mm×100 mm, 1.7 μm) with the mobile phase of 0.1% formic acid aqueous solution (A)-acetonitrile (B) for gradient elution (0-15 min, 10%-22%B; 15-20 min, 22%B; 20-25 min, 22%-44%B; 25-35 min, 44%-50%B; 35-40 min, 50%-60%B; 40-55 min, 60%-85%B), the flow rate was 0.15 mL·min<sup>-1</sup>, the injection volume was 3 μL and the column temperature was 30 ℃. HRMS was equipped with electrospray ionization (ESI) and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan (Full MS/dd-MS<sup>2</sup>). Compound Discoverer 3.0 software combined with mzCloud, mzVault, ChemSpider databases and HRMS database of components in traditional Chinese medicine were used to analyze and identify the collected data by HRMS, based on accurate relative molecular mass, retention time and characteristic ion fragmentation of the compounds, as well as literature information and relevant reference materials. Result:A total of 51 chemical constituents were identified in <italic>C</italic>.<italic> chinensis</italic> inflorescence, including 16 alkaloids, 14 flavonoids, 7 phenylpropanoids, 7 organic acids and 7 others. Among them, 10 components [berberine, palmatine, coptidine, rutin, quercetin, isoquercitrin, chlorogenic acid, cryptochlorogenic acid,<italic> D</italic>-(-) quinic acid and <italic>D</italic>-proline] were unambiguously identified by comparing with reference standards. Conclusion:The established UPLC-Q-Orbitrap HRMS can be used to accurately analyze and identify chemical constituents of <italic>C. chinensis</italic> inflorescence. A total of 41 chemical constituents are reported from <italic>C. chinensis</italic> inflorescence for the first time and 6 alkaloids are found from the <italic>C. chinensis</italic> for the first time. These findings can provide methodological reference and experimental basis for the basic research of quality evaluation and efficacy materials of <italic>C. chinensis</italic> inflorescence, and lay a foundation for its further development and utilization.
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Chinese herbal medicines (CHMs) are one of the important bioresources of medicine, which works by unlocking nature's ability to prevent diseases and recover from illnesses. Recently, it has ascended to the world stage and become a global icon. Nowadays, a considerable of researches have focused on the quality evaluation of CHMs. However, it is difficult to meet the reasonable needs of human beings for safe drug use to evaluate the quality of a huge number of inferior goods for the CHMs contaminated by pesticides and heavy metals. Hence to explore an eligible medicinal plant cultivation pattern, which can provide high quality CHMs sustainably, is most promising. This review analyzed the situation and characteristics of medicinal plant resources in different periods, including wild-harvested and cultivated resources during different stages, putting forward that ecological cultivation must be the way to develop medicinal plant cultivation and to obtain high quality CHMs.
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Radiotherapy for liver cancer can affect the level of autophagy in cells, and effective autophagy regulation can increase the radiosensitivity of liver cancer cells.Saikosaponin-d (SSd) is an effective active ingredient extracted from traditional Chinese medicine Bupleurum. We have confirmed previously in vitro and in vitro experiments that SSd can significantly induce apoptosis of liver cancer cells, increase the radiosensitivity of liver cancer cells.This study explored the role of autophagy in SSd-mediated radiosensitivity of liver cancer cells. MTT and clone formation experiments showed that radiation can inhibit the proliferation of hepatoma cells and reduce the colony formation of hepatoma cells. After the addition of SSd, the inhibitory effect of radiation on the proliferation and clonal formation of hepatoma cells was further enhanced. However, the addition of the autophagy inhibitor chloroquine or mTOR agonist can partially reverse the inhibitory effect of the combined treatment of SSd with radiation on the proliferation of hepatoma cells. Similarly, transmission electron microscopy and laser confocal microscopy showed that after the addition of SSd, the number of radiation-induced autophagosomes increased significantly in hepatoma cells and the intervention of mTOR agonist can reduce the formation of autophagosomes in hepatoma cells.In addition,Western blot analysis presented that radiation significantly increased LC3-II levels. Especially when SSd is added, LC3-II levels is further increased. Our data indicate that SSd can inhibit the growth of liver cancer cells and enhance cell radiosensitivity by inducing autophagy formation.
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Objective: Flower herbs are an important category of traditional Chinese medicinal materials, some of which are used as healthcare tea in folk. However, the increasing adulteration of medicinal herbs is threatening consumer safety. The adulteration of flower herbs and their healthcare tea products in the market were investigated. Methods: A total of 33 flower herb samples from several retail pharmacies in China were randomly collected and 27 flower healthcare tea samples were purchased online. They were identified using ITS2-based Traditional Chinese Medicine Database (TCMD). Additionally, Lonicerae Japonicae Flos and the adulterants were compared in the ITS2 secondary structures. Results: There were one adulterant (Inulae Flos) in flower herb materials and eight adulterants in healthcare tea samples. Inula linariifolia was an adulterate species of Inulae Flos, Robinia pseudoacacia was of Sophorae Flos, and Lonicera macranthoides was of Lonicerae Japonicae Flos. Sophorae Flos and Lonicerae Japonicae Flos were two healthcare tea products with high adulteration rates. Conclusion: The TCMD is powerful tool to identify flower herbs and the adulterants that frequently occurred in the flower herb market, especially online shops.
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Rhizoma Bletillae is in high demand as a traditional Chinese medicine, and the natural resources have been severely damaged due to excessive exploitation. Because Bletilla striata seeds are small and have no endosperm, the seed germination rate is low in natural conditions. Traditional division propagation could not meet the demands of large-scale cultivation. Tissue culture can provide many seedlings in a short time and is more effective and convenient than other methods. Most studies on tissue culture of B. striata selected seeds as explants. This review summarized the processes of the aseptic seed germination pathway. It included such stages as seed germination, proliferation of clusters of buds, induction of rooting and transplanting of seedlings. Influential factors as well as optimum combination and concentration of the plant growth regulators of each stage were also summarized. The further research on tissue culture of B. striata was also prospected.
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ICP-MS was used to determine the contents of five heavy metals such as lead (Pb), cadmium (Cd), mercury (Hg), arsenic (As) and copper (Cu) in Chrysanthemum indicum and soil samples. Pearson correlation and path analysis were used to analyze the effects of latitude and longitude and altitude on the content of heavy metal elements in Ch. indicum. To explore the relationship between geographical location of heavy metal content in Ch. indicum and the content of heavy metal elements in soil. The results showed that the Ch. indicum had high enrichment of Cd and had a synergistic effect on the absorption of five heavy metal elements. Elevation directly affected the accumulation of Pb and Cd in Ch. indicum and was a positive effect. The direct positive effect of latitude acts on the accumulation of Hg and Cu in Ch. indicum. The effect of longitude on the Cd content was the negative effect of longitude. The geographical location had different effects on the accumulation of heavy metal elements in Ch. indicum. There was a certain interaction between direct and indirect effects. Ch. indicum had different absorption and enrichment characteristics for different heavy metal species. When controlling the heavy metal content and medicinal material quality of Ch. indicum, we must fully consider the influence of geographical location on the quality of medicinal materials, select suitable geographic regions to carry out acquisitions, and choose a good ecological environment. Regional establishment of Chinese herbal medicine planting bases, or adopting certain agricultural measures to reduce the absorption of heavy metal elements in Ch. indicum herbs, while conducting research on chemical composition, strengthening the research on the effective forms of heavy metals, and paying attention to the safety of medicinal herbs.
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Abundant evidence supports the key role of ultraviolet radiation (UVR) in skin cancer development. The human skin, especially the epidermal layer, is the main defense against UV radiation. Baicalin is a major bioactive component of Scutellaria baicalensis Georgi, a plant which has been found to exhibit antitumor activity. The anticarcinogenic mechanism of baicalin is not completely understood. We have reported that baicalin inhibited UVB-induced photo-damage and apoptosis in HaCaT cells (human skin keratinocytes). The aim of the present study is to investigate the cellular gene targets responsible for baicalin's antitumor activity by performing two-dimensional electrophoresis liquid chromatography-mass spectrometry/mass spectrometry (2-DE LC-MS/MS) with HaCaT cells following UVB and baicalin exposure. Two-DE for protein separation was performed, followed by matrix-assisted laser desorption/ionization mass spectrometry and database searches. Nucleophosmin (NPM)-specific siRNA was designed and synthesized, and the small interfering RNA was transfected into skin squamous cancer A431 cells to knockdown the NPM expression. Proliferation and cell cycle status were assessed by CCK8 and flow cytometric analyses, respectively. We have identified 38 protein spots that are differentially expressed in HaCaT cells exposed to baicalin and/or UVB irradiation These proteins are involved in detoxification, proliferation, metabolism, cytoskeleton and motility. In particular, we found several proteins that have been linked to tumor progression and resistance, such as NPM. Baicalin treatment reduced the cellular proliferation rate and induced arrest during the S-phase of the cell cycle in A431 cells. NPM1 silencing significantly enhanced the effect of baicalin. Our data indicated that baicalin results in the significant inhibition of tumor growth in the A431 cell line, which may be associated with the regulation of the NPM gene expression.
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Antineoplásicos Fitogênicos , Flavonoides/genética , Flavonoides/farmacologia , Fitoterapia , Proteômica , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologia , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Transformação Celular Neoplásica/efeitos dos fármacos , Flavonoides/uso terapêutico , Humanos , Terapia de Alvo Molecular , Nucleofosmina , Interferência de RNA/efeitos dos fármacos , RNA Interferente Pequeno , Scutellaria baicalensis/química , Neoplasias Cutâneas/genética , Células Tumorais CultivadasRESUMO
Objective To seek for the differentially-expressed proteins in patients with kidney-yin deficiency syndrome and to screen out the specific proteins,so as to provide evidence for the establishment of objective standard of kidney deficiency syndrome of traditional Chinese medicine (TCM).Methods Five patients with typical kidney-yin deficiency syndrome and 6 normal healthy volunteers were enrolled into the study.Plasma proteins in both groups were detected by antibody chip,and then the plasma proteins profile was compared and analyzed.Results A total of 25 differentially-expressed proteins between kidney-yin deficiency group and normal control group were found,of which 2 were up-regulated and 23 were down-regulated.Conclusion The differentially-expressed proteins in patients with kidney-yin deficiency syndrome are mainly related to immune disorder,protein biosynthesis,metabolism,oxidative stress,cell apoptosis,signal transduction,and so on.
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Taiwan is known for its high quality oolong tea. Because of high consumer demand, some tea manufactures mix lower quality leaves with genuine Taiwan oolong tea in order to increase profits. Robust scientific methods are, therefore, needed to verify the origin and quality of tea leaves. In this study, we investigated whether two-dimensional gel electrophoresis (2-DE) and nanoscale liquid chromatography/tandem mass spectroscopy (nano-LC/MS/MS) coupled with a two-layer feature selection mechanism comprising information gain attribute evaluation (IGAE) and support vector machine feature selection (SVM-FS) are useful in identifying characteristic proteins that can be used as markers of the original source of oolong tea. Samples in this study included oolong tea leaves from 23 different sources. We found that our method had an accuracy of 95.5% in correctly identifying the origin of the leaves. Overall, our method is a novel approach for determining the origin of oolong tea leaves.
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Eletroforese em Gel Bidimensional/métodos , Folhas de Planta/química , Proteômica/métodos , Chá/química , Eletroforese em Gel Bidimensional/normas , Folhas de Planta/genética , Taiwan , Espectrometria de Massas em Tandem/métodos , Chá/genéticaRESUMO
Profile distribution characteristics of organic carbon (C), total nitrogen (N), total phosphorus (P) and total sulfur (S) were studied in two typical marshes including Carex lasiocarpa marsh and Phragmites australis marsh in the Sanjiang Plain. Sedimentary characteristics of typical mashes were analyzed. The results showed that vertically these soil chemical elements also varied, showing obvious stratification and enrichment. In a soil profile, soil organic C under both vegetation communities gradually decreased; soil total N first increased and then decreased under both; total P under Carex lasiocarpa first decreased and then increased, whereas it decreased with the increasing depth under Carex lasiocarpa; total S was reduced with increasing depth under both marshes. Total N, total P and total S were all strongly correlated with soil organic C (P < 0.01); soil organic C was strongly correlated with bulk density (P < 0.01). Our study also illustrated that the vegetation types had different influences for organic C, total N, total P and total S of the marsh profiles. Environmental 137Cs and 210Pb dating techniques were applied to determine recent sedimentation rates, and the constant rate of supply (CRS) was applied to deduce the age of sediment core, and the results showed that the mean sedimentation rate was 0.33 cm x a(-1), and the sedimentation fluxes ranged 0.03-0.48 g x (cm2 x a)(-1) [Mean = 0.29 g x (cm2 x a) -1].
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Sedimentos Geológicos/química , Nitrogênio/análise , Fósforo/análise , Enxofre/análise , Áreas Alagadas , Carex (Planta) , China , Poaceae , Solo/químicaRESUMO
Ultraviolet A (UVA) radiation contributes to skin photoaging. Baicalin, a plant-derived flavonoid, effectively absorbs UV rays and has been shown to have anti-oxidant and anti-inflammatory properties that may delay the photoaging process. In the current study, cultured human skin fibroblasts were incubated with 50 µg/ml baicalin 24 hours prior to 10 J/cm(2) UVA irradiation. In order to examine the efficacy of baicalin treatment in delaying UVA-induced photoaging, we investigated aging-related markers, cell cycle changes, anti-oxidant activity, telomere length, and DNA damage markers. UVA radiation caused an increased proportion of ß-Gal positive cells and reduced telomere length in human skin fibroblasts. In addition, UVA radiation inhibited TGF-ß1 secretion, induced G1 phase arrest, reduced SOD and GSH-Px levels, increased MDA levels, enhanced the expression of MMP-1, TIMP-1, p66, p53, and p16 mRNA, reduced c-myc mRNA expression, elevated p53 and p16 protein expression, and reduced c-myc protein expression. Baicalin treatment effectively protected human fibroblasts from these UVA radiation-induced aging responses, suggesting that the underlying mechanism involves the inhibition of oxidative damage and regulation of the expression of senescence-related genes, including those encoding for p53, p66(Shc) and p16.
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Senescência Celular/genética , Senescência Celular/efeitos da radiação , Fibroblastos/efeitos da radiação , Flavonoides/farmacologia , Envelhecimento da Pele/genética , Envelhecimento da Pele/efeitos da radiação , Pele/citologia , Pele/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Anti-Inflamatórios não Esteroides , Antioxidantes , Ciclo Celular/efeitos dos fármacos , Células Cultivadas , Inibidor p16 de Quinase Dependente de Ciclina , Dano ao DNA/efeitos dos fármacos , Fibroblastos/citologia , Fibroblastos/metabolismo , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Humanos , Proteínas de Neoplasias/metabolismo , Proteínas Adaptadoras da Sinalização Shc/metabolismo , Proteína 1 de Transformação que Contém Domínio 2 de Homologia de Src , Homeostase do Telômero/efeitos dos fármacos , Fator de Crescimento Transformador beta1/metabolismo , Proteína Supressora de Tumor p53/metabolismoRESUMO
<p><b>OBJECTIVE</b>To study the effect of extract of Radix Tetrastigma hemsleyani on the proliferation and apoptosis of human lung carcinoma H1299 cells, and to explore its mechanisms. METHODS H1299 cells were treated with the extract of Radix Tetrastigma hemsleyani in different concentrations at different time points. Its inhibition on H1299 cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. The morphology of the H1299 cell was observed by inverted microscope. Changes of apoptosis were observed by Hoechst33258 methods. The apoptosis rate was detected by flow cytometry. Expression changes of apoptosis-related proteins pro-caspase-3, pro-caspase-9, cle-caspase-3, cle-caspase-9, and poly-ADP-ribose polymerase (PARP) were detected by Western blot.</p><p><b>RESULTS</b>Compared with the control group, the inhibition rate of H1299 cells increased after acted by 0.5, 1, 5, and 10 mg/mL extract of Radix Tetrastigma hemsleyani (P < 0.05, P < 0.01). The longer the acting time, the higher the inhibition rate (P < 0.01). Under inverted microscope, typical morphological changes could be seen and the number of H1299 cells was reduced. Under fluorescence microscope, dark stained nucleus and formed apoptotic body could be observed. Results of flow cytometry showed that the apoptosis rate was obviously dose-effect correlated with the concentration of extract of Radix Tetrastigma hemsleyani. Results of Western blot indicated that compared with the control. group, the protein expression of pro-caspase-3, pro-caspase-9, and PARP were down-regulated and that of cle-caspase-3, cle-caspase-9, and cle-PARP were up-regulated by 5 and 10 mg/mL extract of Radix Tetrastigma hemsleyani (P < 0.05).</p><p><b>CONCLUSIONS</b>Extract of Radix Tetrastigma hemsleyani had obvious effect in inhibiting the proliferation and inducing apoptosis of human lung carcinoma H1299 cells, which might be achieved by activating the expression of caspase protein.</p>