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Resumen: El gel de Aloe vera es considerado una fuente natural de múltiples beneficios, originados por la acción combinada de vitaminas, aminoácidos, compuestos fenólicos, enzimas, minerales, ácidos orgánicos, lípidos y carbohidratos, que se relacionan con la mejora de enfermedades neuro-degenerativas como Alzheimer. Los ensayos in vitro e in silico permiten confirmar e identificar posibles beneficios de esta planta y sus compuestos en enfermedades. El objetivo del presente trabajo fue evaluar la actividad antioxidante del gel de A. vera y mediante análisis in silico, establecer el potencial terapéutico de sus compuestos bioactivos en la enfermedad de Alzheimer. Se obtuvieron hojas de A. vera, de las que se extrajo el gel, retirando el exocarpio, se liofilizó y almacenó hasta su uso. Se caracterizó la capacidad antioxidante, se cuantificaron los compuestos fenólicos y flavonoides y se analizó la relación que existe entre los parámetros mediante correlación de Pearson. Mediante análisis in silico se evaluó el potencial de interacción de 8 compuestos del gel con la proteína gamma secretasa. El gel de A. vera obtuvo alta capacidad antioxidante por ABTS, DPPH, radical OH y poder reductor, usando bajas concentraciones para inhibir el 50 % de los radicales, y correlaciones positivas con fenoles totales y flavonoides. En el estudio in silico el compuesto que presentó mejor unión con gamma secretasa fue aloe-emodina, con menor energía libre de unión y menor concentración de constante de inhibición, sugiriendo su potencial uso como coadyuvante en el tratamiento de la enfermedad de Alzheimer.
Abstract: Aloe vera gel is considered a natural source of multiple benefits, originated by the combined action of vitamins, amino acids, phenolic compounds, enzymes, minerals, organic acids, lipids and carbohydrates, which are related to the improvement of neuro-degenerative diseases such as Alzheimer's. In vitro and in silico tests allow us to confirm and identify possible benefits of this plant and its compounds in diseases. The objective of the present study was to evaluate the antioxidant activity of A. vera gel and, through in silico analysis, to establish the therapeutic potential of its bioactive compounds in Alzheimer's disease. A. vera leaves were obtained, from which the gel was extracted, removing the exocarp, lyophilized and stored until use. The antioxidant capacity was characterized, the phenolic compounds and flavonoids were quantified, and the relationship between the parameters was analyzed using Pearson correlation. The interaction potential of 8 compounds in the gel with the gamma secretase protein was evaluated through in silico analysis. The A. vera gel obtained high antioxidant capacity due to ABTS, DPPH, OH radical and reducing power, using low concentrations to inhibit 50 % of the radicals, and positive correlations with total phenols and flavonoids. In the in silico study, the compound that showed the best binding with gamma secretase was aloe-emodin, with lower binding free energy and lower inhibition constant concentration, suggesting its potential use as an adjuvant in the treatment of Alzheimer's disease.
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Objective To explore the possible mechanism of emodin in inhibiting proliferation,migration,and invasion of AGS cells and in suppressing the expressions of YAP1 and FOXD1.Methods Normal gastric cell GES-1 and gastric cancer cell AGS were cultured with different concentrations of emodin.CCK8 test,scratch test and Transwell assay were used to verify changes in the biological phenotype of AGS cells.TCGA database was applied to analyze expressions of HK2,YAP1 and FOXD1 in gastric cancer tissues and normal gastric tissues.Western blotting method was used to detect the impacts of emodin on HK2,YAP1 and FOXD1 proteins in AGS cells.Exogenous pyruvic acid was added to verify the changes in YAP1 and FOXD1.Results The IC50 of emodin was significantly higher in GES-1 cells than in AGS cells(P<0.05).CCK8 proliferation test,scratch test,and Transwell assay showed that emodin significantly inhibited the biological abilities of AGS(P<0.05 for comparisons).Analysis on the TCGA bioinformatics database found that the expression of key enzymes HK2 in the glycolysis pathway and oncogenes YAP1 and FOXD1 was significantly higher in gastric cancer tissues than in normal gastric tissues(P<0.05 for comparisons).Emodin significantly inhibited the protein expressions of key glycolytic enzymes HK2 and oncogenes YAP1 and FOXD1(P<0.05 for comparisons).With supplement of exogenous glycolytic metabolite pyruvate,the protein expressions of oncogenes YAP1 and FOXD1 significantly increased(P<0.05 for comparisons).Conclusions Emodin has a significant pharmacological inhibitory effect on gastric cancer AGS cells,markedly suppressing their biological phenotype.Emodin not only significantly inhibits the key enzyme HK2 in glycolysis metabolism,but also the protein expressions of oncogenes YAP1 and FOXD1.With the addition of exogenous pyruvate to enhance the glycolytic metabolic pathway,the protein expressions of oncogenes YAP1 and FOXD1 significantly increased.The above results suggest a close association of YAP1 and FOXD1 with glycolytic metabolism.Emodin may inhibit oncogenes YAP1 and FOXD1 through the glycolytic metabolism of gastric cancer AGS cells.
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Objective To study whether emodin,a natural product,can affect the level of histone acetylation in HpG2 hepatocellular carcinoma cells(HCC),and then accelerate the occurrence of pyroptosis and apoptosis in hepatocellular carcinoma cells,so as to provide a new target for the treatment of liver cancer.Methods CCK-8 method was used to detect the effect of different concentrations of emodin on the viability of HpG2 cells.Bioinformatics was used to analyze histone acetylation-related genes in patients with liver cancer in TCGA database.The correlation between the candidate gene lysine acetyltransferase 2A(KAT2A)and the apoptosis pathway was verified.qPCR method was used to detect the mRNA level of KAT2A in HepG2 cells and L02 cells.The effects of emodin on histone acetyltransferase(HAT)and histone deacetyltransferase(HDAC),interleukin 1β(IL-1β)and interleukin 18(IL-18)in HpG2 cells were detected by ELISA.The effect of emodin on the apoptosis of liver cancer cells was detected by flow cytometry.The expression level of cell apoptosis,pyroptosis-associated protein B lymphocytoma-2(Bcl-2),Bcl-2-related X protein(Bax),NOD-like receptor thermal protein domain-related protein 3(NLRP3),Caspase-1,Gasdermin family member DN terminal(GSDMD-N)and KAT2A were detected by Western blot assay.Results Emodin could reduce the activity of HpG2 cells,and the confidence interval of IC5095%was 58.12-66.52μmol/L.Compared with normal liver tissue,the expression of histone acetylation related gene mRNA was increased in HCC tissue,and the change of KAT2A was the highest[log2(Fold Change)=2.010,P＜0.01].In HCC tissue,the expression of KAT2A mRNA was negatively correlated with apoptosis pathway(rs=-0.230,P＜0.01).Compared with L02 cells,the expression of KAT2A mRNA in HepG2 was higher(P＜0.05).Compared with the control group,expression levels of HAT and HDAC decreased in the 60μmol/L emodin intervention group,expression levels of IL-18 and IL-1β increased,the apoptosis rate increased,expression levels of KAT2A and BAX decreased,and expression levels of Bcl-2,NLRP3,GSDMD-N and Caspase-1 increased(P＜0.05).Conclusion Emodin could inhibit the viability of hepatoma cells,accelerate apoptosis and pyroptosis,and its mechanism may be related to the regulation of KAT2A expression.
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BACKGROUND:Previous studies have shown that silent mating-type information regulator 2 homolog 1(SIRT1)-mechanistic target of rapamycin(mTOR)signaling plays an important role in the progression of osteoarthritis.Emodin has a protective effect on osteoarthritic chondrocytes. OBJECTIVE:To investigate the effects of emodin on the proliferation,apoptosis and oxidative stress of osteoarthritic chondrocytes based on the SIRT1-mTOR signaling. METHODS:Rat chondrocytes were isolated and cultured in vitro.Osteoarthritic chondrocyte model in vitro was induced by 10 ng/mL interleukin-1β.Cell counting kit-8 method was used to determine the viability of rat chondrocytes treated with 0,20,40,80,120,160 μmol/L emodin,and the appropriate concentration of emodin was selected.Rat chondrocytes isolated and cultured in vitro were randomly divided into control group,model group,low-dose emodin group,high-dose emodin group,EX527 group,and high-dose emodin+EX527 group.In vitro osteoarthritis model was constructed by induction of 10 ng/mL interleukin 1β in all groups except the control group.The cells in the latter four groups were correspondingly treated with emodin or/and EX527.The proliferation and apoptosis of chondrocytes in each group were detected by cell counting kit-8,Edu staining and flow cytometry respectively.The relative content of reactive oxygen species and the levels of malondialdehyde,superoxide dismutase,catalase,and glutathione peroxidase in chondrocytes of rats in each group were measured with the kit.The expression of proteins related to cell matrix degradation,apoptosis and the SIRT1-mTOR pathway-related proteins in each group were detected by western blot. RESULTS AND CONCLUSION:Compared with the control group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the model group were decreased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were increased(P<0.05).Compared with the model group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the low-and high-dose emodin groups were increased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were decreased(P<0.05).Compared with the low-and high-dose emodin groups,the indexes of EX527 group showed the opposite trend(P<0.05).Compared with the high-dose emodin group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the high-dose emodin+EX527 group were decreased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were increased(P<0.05).To conclude,emodin can inhibit oxidative stress of osteoarthritic chondrocytes by activating the SIRT1-mTOR signaling,thereby promoting chondrocyte proliferation and reducing apoptosis.
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Objective To study the cGAS/STING signaling pathway and investigate the potential effect of emodin(EMD)on autophagy of human rheumatoid arthritis fibroblast synovial cells(MH7A).Methods CCK-8 method was used to detect MH7A cell proliferation,and the experimental concentration of EMD was screened according to cell survival rate.Then,autophagy inhibitor 3-MA was added to further verify the effect of EMD on autophagy.Autophagy of MH7A cells was detected via the monodansylcadaverine staining method.Protein expression levels of cGAS,STING,p-STING,LC3-Ⅰ,LC3-Ⅱ,P62 and Beclin-1 were detected by Western blot.Results Monodansylcadaverine staining indicated that EMD enhanced the autophagy of MH7A cells.Western blot indicated that EMD decreased the expression of autophagy related proteins cGAS,STING,p-STING and P62,and increased that of LC3-Ⅱ and Beclin-1 in MH7A cells.After addition of the autophagy inhibitor 3-MA,the expression of P62 protein in MH7A cells increased,while that of LC3-Ⅱ and Beclin-1 decreased.Conclusions EMD may accelerate autophagy and inhibit MH7A cell proliferation by down-regulating cGAS/STING signaling pathway proteins.
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OBJECTIVE To extract and isolate the four chemical components of Yao medicine Ventilago leiocarpa， and to conduct identification and content determination for them. METHODS The chemical components of V. leiocarpa were separated and purified by solvent extraction， extraction， silica gel column chromatography and preparative liquid chromatography； then the chemical structures of four isolated compounds were identified based on their spectral data. The contents of four components were determined by high performance liquid chromatography（HPLC）-quantitative analysis of multi-components by single-marker （QAMS） method， with the following chromatographic conditions： chromatographic column was Echway GowonTM C18 （250 mm× 4.6 mm， 5 μm）. The mobile phase was acetonitrile-0.1% phosphoric acid for gradient elution； the detection wavelength was 269 nm， and the column temperature was 25 ℃ . Using emodin as internal reference， the relative correction factors （fi/s） between emodin and the other 3 components were established and used to calculate the content. At the same time， the content of each component was calculated with the external standard method （ESM）， and the differences between these two methods were compared. RESULTS Four compounds were isolated from V. leiocarpa， and they were identified as emodin， frangulin A， pleuropyrone A， emodin-8-O-β-D-glucoside. The result of HPLC-QAMS showed that the fi/s of pleuropyrone A， emodin-8-O-β-D- glucoside and frangulin A were 1.147 2， 0.874 7 and 0.644 4， respectively. The content of these four components was measured as a good linearity （r≥0.999 6）； relative standard deviation （RSD） of precision， stability and reproducibility tests were all lower than 2.00%， and average recoveries were E-mail：dearhuangjianyou@126.com 99.41%-100.46%（RSD≤2.05%）. There was no significant difference between QAMS method and ESM （RSD＜3.00%）. CONCLUSIONS Emodin， frangulin A， pleuropyrone A and emodin- 8-O-β-D-glucoside are isolated from V. leiocarpa； among them， the last three components are all isolated from for the first time. The established HPLC-QAMS method is accurate and reliable for the determination of 4 components in V. leiocarpa， and can used for quality control of V. leiocarpa.
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The cis-emodin-emodin dianthrone (compound 1) and trans-emodin-emodin dianthrone (compound 2) were extracted from Polygonum multiflorum Thunb. The protective effect and mechanism of compound 1 and compound 2 (emodin-emodin dianthrones) on acute liver injury induced by concanavalin A (ConA) in ICR mice was first investigated. The results indicated that emodin-emodin dianthrones at 1 mg·kg-1 significantly reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) level (P < 0.05). Emodin-emodin dianthrones also improved liver histopathological damage in liver-injured mice. The level of Bcl-2-associated X protein (Bax) mRNA in liver was significantly reduced by 1 mg·kg-1 of emodin-emodin dianthrones, while the level of B-cell lymphoma-2 (Bcl-2) mRNA expression was significantly increased (P < 0.05). The protective activity of compounds 1 and 2 against hepatocyte injury was further evaluated by hydrogen peroxide (H2O2)-induced hepatocyte injury. Compounds 1 and 2 significantly inhibited H2O2-induced hepatocyte injury and reduced the levels of ALT, AST, alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) in cell culture. Compounds 1 and 2 also significantly improved the cell survival rate and decreased H2O2-induced oxidative stress in hepatocytes. Compound 1 (0.5 µmol·L-1) significantly increased the enzymatic activity of superoxide dismutase (SOD) in hepatocytes (P < 0.01), and 0.5 µmol·L-1 of compound 2 significantly decreased the intracellular reactive oxygen species (ROS), increased SOD enzyme activity, and glutathione (GSH) content (P < 0.01). Compounds 1 and 2 at 0.5 µmol·L-1 also inhibited hepatocyte apoptosis by increasing the protein expression ratio of Bcl-2/Bax (P < 0.05) and decreasing the protein expression ratio of cleaved caspase-3 and pro caspase-3 (P < 0.05). This study indicates that the emodin-emodin dianthrones from Polygonum multiflorum Thunb. have liver-protective activity. Compounds 1 and 2 exerted hepatoprotective effects by inhibiting apoptosis and oxidative stress. The study provides an important material basis for the hepatoprotective effect of commonly used amounts of Polygonum multiflorum Thunb.
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This study was designed to investigate the effect of emodin(EMO)on endoplasmic reticulum stress(ERS)in alveolar macrophages(AMs)of rats with severe acute pancreatitis(SAP)and the underlying mechanism.Forty rats were recruited and randomized into four groups:sham-operation(SO)group,SAP group,4-phenylbutyric acid(4-PBA)group and EMO group.The SAP model was established by retrograde injection of 5%sodium taurocholate into the biliopancreatic duct.HE staining was performed to observe the pathological changes in the pancreas and lung;the wet/dry weight ratio of lung tissue was calculated.Arterial partial pressure of the oxygen(PaO2)and carbon dioxide(PaCO2)were measured;the serum amylase activity and the levels of inflammatory factors TNF-α and IL-6 were evaluated.TUNEL staining was used to determine the cell apoptosis rate of lung tissue;Western blot was used to detect the protein expression levels of GRP78,ATF6,CHOP,and Caspase-12.Moreover,rat AMs(NR8383)were signed into control(CON)group,lipopolysaccharide(LPS)group,4-phenylbutyric acid(4-PBA)group and EMO group in vitro.Glutathione(GSH)and malondialdehyde(MDA)levels in the cell medium were measured,as were the protein expression levels of GRP78,ATF6,CHOP,and Caspase-12 in AMs.For experiments in vivo,compared with the SO group,the pathological score of pancreatic and lung in SAP rats was increased(P<0.05),the levels of serum amylase activity,IL-6 and TNF-α were increased(P<0.05),the wet/dry weight ratio and apoptosis rate of lung tissue were increased(P<0.05),PaO2 was decreased,PaCO2 was increased(P<0.05),and the protein expression of GRP78,ATF6,CHOP and Caspase-12 were increased in lung tissue(P<0.05).Compared with the SAP group,these indexes mentioned above in the 4-PBA and EMO groups were reversed(P<0.05).For experiments in vitro,compared with the CON group,GSH levels were decreased,and MDA levels were increased in the cell medium of LPS group(P<0.05),and the expression of GRP78,ATF6,CHOP and Caspase-12 proteins were upregulated(P<0.05).Compared with the LPS group,these indexes mentioned above in in cell medium of the 4-PBA and EMO groups were reversed(P<0.05).In conclusion,the protective effect of EMO on pancreatic and lung injury in SAP rats may be closely related to the inhibition of ATF6/CHOP pathway-induced inflammation and oxidative stress in AMs.
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Objective To investigate the effect and possible mechanism of Emodin on Doxorubicin(DOX)-induced cardiotoxicity.Methods H9C2 cells were preincubated with low,medium and high doses(5,15 and 25 μmol·L-1)of Emodin for 8 h,and then cardiomyocytes were treated with 2.5 μmol·L-1 DOX for 16 h to establish a cardiotoxic injury model.MTT assay was used to detect the cell survival rate.The levels of cardiac troponin(cTnT),lactate dehydrogenase(LDH),creatine kinase(CK),superoxide dismutase(SOD),reactive oxygen species(ROS),malondialdehyde(MDA),reduced glutathione(GSH),glutathione peroxidase(GSH-Px)and the activities of Caspase-3 and Caspase-9 in H9C2 cells were detected by kit;RT-qPCR was used to detect miR-29a-5p and vascular endothelial growth factor-B(VEGFB)expression levels.The expression levels of VEGFB protein and other apoptosis-related factors were detected by Western blot.After transfection of miR-29a-5p mimics and inhibitors,the targeting of VEGFB to miR-29a-5p was detected by RT-qPCR and Western blot.H9C2 cells were transfected with miR-29a-5p mimics,Under the dose of Emodin 15 μmol·L-1,the activity of H9C2 cells was detected by MTT method,and the expression levels of miR-29a-5p and VEGFB were detected by RT-qPCR and Western blot.Results Compared with the DOX group,Emodin low,medium and high dose groups significantly increased the activity of H9C2 cells(P<0.05,P<0.01),and decreased the levels of cTnT,LDH and CK(P<0.05,P<0.01);RT-qPCR indicated that the expression level of miR-29a-5p decreased significantly and the expression level of VEGFB increased significantly(P<0.05,P<0.01);The levels of ROS and MDA decreased significantly,and the levels of GSH and GSH-Px increased significantly(P<0.05,P<0.01);TUNEL positive cells decreased significantly,and the activities of Caspase-3 and Caspase-9 decreased significantly.Western blot showed that the expression level of VEGFB increased significantly,the expression level of apoptosis-related factor Bax decreased significantly,and the expression levels of Bcl-2 and Bcl-xl increased significantly(P<0.05,P<0.01).After transfection with miR-29a-5p mimics and inhibitors,the expression of VEGFB decreased and increased respectively(P<0.05,P<0.01).After transfection of miR-29a-5p mimics and treatment with Emodin 15 μmol·L-1 of H9C2 cells,the significant decrease in the activity of H9C2 cells induced by DOX was reversed,and the significant decrease in the expression level of VEGFB was also reversed(P<0.05,P<0.01).Conclusion Emodin significantly reduces DOX-induced cardiotoxicity by regulating miR-29a-5p/VEGFB-mediated cardiomyocyte apoptosis.
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Objective:To investigate the effects of emodin on proliferation, adhesion, migration, c-kit mRNA and protein expression of human epidermal melanocytes.Methods:Human epidermal melanocytes cultured in vitro were treated with different concentrations of emodin, a blank control group (containing medium + human epidermal melanocytes) and an experimental group (containing medium + human epidermal melanocytes + different concentrations of emodin) were set up. Cell proliferation was measured by CCK-8 method, cell adhesion was measured by microplate assay, cell migration was measured by Transwell membrane assay, and c-kit mRNA expression was measured by reverse transcription-polymerase polymerase chain reaction; Western blot was used to detect the expression of c-kit protein.Results:Compared with the blank control group, emodin decreased the proliferation, adhesion and migration of human epidermal melanocytes, the difference was statistically significant ( F=391.48, P<0.0001; F=10.93, P=0.003; F=7.75, P=0.009). Compared with the blank control group, emodin in the experimental group had a bidirectional effect on the expression of c-kit mRNA and protein. High concentration of emodin inhibited the expression of c-kit mRNA and protein, and the expression of c-kit mRNA and protein was significantly increased by Emodin ( F=11.491, P=0.003; F=2155.11, P<0.001). Conclusions:Emodin inhibits the proliferation, adhesion and migration of human epidermal melanocytes, high concentration of emodin inhibits the expression of c-kit mRNA and protein, and low concentration of emodin promotes the expression of c-kit mRNA and protein, and the results provide a basis for the clinical application of emodin in pigmented dermatosis.
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@#As an active hydroxyanthraquinone ingredient, emodin is abundant in Chinese medicine herbs, such as Rheum palmatum, Polygonum cuspidatum and Polygonum multiflorum.Modern pharmacological studies have shown that emodin has a variety of pharmacological activities including anti-tumor, anti-inflammatory and immunoregulatory, antibacterial and anti-viral effects, myocardial protection, neuroprotection, renal protection, bone protection, antifibrosis and so on, which indicate its high medicinal value and broad application prospects.This article aims to summarize the progress in the pharmacological activity and mechanism of action of emodin published in domestic and international journals over the last 5 years and highlight the potential targets and molecular signaling pathways linked with emodin, so as to provide some clues and references for further development and clinical application of emodin.
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AIM: To investigate the injury of emodin (EMO) in reduce of glomerular mesangial cells (MCs) in lupus nephritis by targeting forkhead protein K2 (FOXK2) through miR-96-5p. METHODS: The contents of 24 h urine protein, serum urea nitrogen (BUN) and serum creatinine (Scr) in MRL / faslpr mice (lupus nephritis group) and MRL / MPJ mice (control group) were detected. MCs were separated, purified and divided into: MCs group (MCs without any treatment), L-EMO group (MCs treated with 10 μmol/L Emodin), M-EMO group (MCs treated with 25 μmol / L Emodin), H-EMO group (MCs treated with 50 μmol / L Emodin), H-EMO + miR-96-5p-NC group (MCs treated with 50 μmol / L Emodin and transfected with miR-96-5p-NC), and H-EMO + miR-96-5p-minic group (MCs treated with 50 μmol/ L Emodin and transfected with miR-96-5p-minic). Double luciferase report experiment was used to verify the targeting relationship between miR-96-5p and FOXK2. The real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-96-5p. Western blot was used to detect the expression of FOXK2 and apoptosis related proteins. The enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors in MCs. cell count kit 8 (CCK-8) was used to determine the activity of MCs. Annexin-V FITC/PI double staining was used to detect apoptosis of MCs. RESULTS: Compared with the control group, 24 h urinary protein content, serum BUN and Scr levels in the lupus nephritis group were significantly increased (P< 0.05). Compared with the MCs group, the miR-96-5p expression, interleukin1β (IL-1β), interleukin6 (IL-6), tumor necrosis factor-α (TNF-α), A450 value and B-lymphoblastoma-2 (Bcl-2) protein in the L-EMO group, M-EMO group and H-EMO group were significantly decreased (P<0.05), the FOXK2 level, cell apoptosis rate, Bcl-2 related X gene (Bax), aspartate specific cysteine proteinase-3 (cleaved Caspase-3) protein levels were significantly increased, respectively (P<0.05), the effect of Emodin was dose-dependent. Compared with the H-EMO group and H-EMO+miR-96-5p-NC group, H-EMO+miR-96-5p-minic group obviously increased the miR-96-5p expression, inflammatory factor levels, A450 value and Bcl-2 protein level (P<0.05), and obviously decreased FOXK2 level and cell apoptosis rate (P< 0.05). CONCLUSION: EMO can reduce the injury of lupus nephritis MCs by down-regulating miR-96-5p and then up-regulating FOXK2.
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OBJECTIVE@#To investigate the effect of emodin on high glucose (HG)-induced podocyte apoptosis and whether the potential anti-apoptotic mechanism of emodin is related to induction of adenosine-monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)-mediated autophagy in podocytes (MPC5 cells) in vitro.@*METHODS@#MPC5 cells were treated with different concentrations of HG (2.5, 5, 10, 20, 40, 80 and 160 mmol/L), emodin (2, 4, 8 µ mol/L), or HG (40 mmol/L) and emodin (4 µ mol/L) with or without rapamycin (Rap, 100 nmol/L) and compound C (10 µ mol/L). The viability and apoptosis of MPC5 cells were detected using cell counting kit-8 (CCK-8) assay and flow cytometry analysis, respectively. The expression levels of cleaved caspase-3, autophagy marker light chain 3 (LC3) I/II, and AMPK/mTOR signaling pathway-related proteins were determined by Western blot. The changes of morphology and RFP-LC3 fluorescence were observed under microscopy.@*RESULTS@#HG at 20, 40, 80 and 160 mmol/L dose-dependently induced cell apoptosis in MPC5 cells, whereas emodin (4 µ mol/L) significantly ameliorated HG-induced cell apoptosis and caspase-3 cleavage (P<0.01). Emodin (4 µ mol/L) significantly increased LC3-II protein expression levels and induced RFP-LC3-containing punctate structures in MPC5 cells (P<0.01). Furthermore, the protective effects of emodin were mimicked by rapamycin (100 nmol/L). Moreover, emodin increased the phosphorylation of AMPK and suppressed the phosphorylation of mTOR. The AMPK inhibitor compound C (10 µ mol/L) reversed emodin-induced autophagy activation.@*CONCLUSION@#Emodin ameliorated HG-induced apoptosis of MPC5 cells in vitro that involved induction of autophagy through the AMPK/mTOR signaling pathway, which might provide a potential therapeutic option for diabetic nephropathy.
الموضوعات
Emodin/pharmacology , AMP-Activated Protein Kinases/metabolism , Podocytes , Caspase 3/metabolism , TOR Serine-Threonine Kinases/metabolism , Signal Transduction , Apoptosis , Sirolimus/pharmacology , Glucose/metabolism , Autophagyالملخص
Purpose: Interstitial cystitis/bladder pain syndrome (IC/BPS) is a devastating urological chronic pelvic pain condition. In search of a potential treatment, we investigated the effect of emodin on IC/BPS inflammation and fibrosis, and explore the potential mechanism. Methods: An experimental model of interstitial cystitis was induced by cyclophosphamide, and human bladder smooth muscle cells were treated with lipopolysaccharide to establish the cell model in vitro. In both models, inflammation- and fibrosis-related indexes were measured after emodin administration. Furthermore, the specific antagonists were used to dig for the mechanisms underlying the response to emodin treatment. Results: Emodin significantly ameliorated management of cystitis, reduced the amount of inflammatory cytokines (tumor necrosis factor-α, monocyte chemoattractant protein-1, interleukin-1ß, interleukin-8, and interleukin-6) in models, as well as reducing the synthesis of fibrosis marker including collagen1, collagen3, vimentin, fibronectin and α-smooth muscle actin. Further mechanism studies demonstrated that emodin inhibited inflammatory reaction and fibrosis through blocking lysine-specific demethylase 6B (JMJD3) expression via JAK/STAT, NF-κB and TGF-ß/SMAD pathways. Conclusions: Our study reveals the critical role of emodin-JMJD3 signaling in interstitial cystitis by regulating inflammation, fibrosis, and extracellular matrix deposition in cells and tissues, and these findings provide an avenue for effective treatment of patients with cystitis.
الموضوعات
Animals , Mice , Fibrosis , Emodin , Cystitis, Interstitial , Inflammationالملخص
Aloe vera, a popular succulent perennial medicinal plant with a wide range of phytochemicals that have shown various pharmacological activities including anti-oxidant, antimicrobial, antidiabetic, wound healing promotion and so on. Acemannan, aloe-emodin, aloin, aloesin, and emodin are widely investigated active constituents that show various pharmacological activities. Thus, the purpose of this review is to highlight previous pharmacological studied conducted in vivo, in vitro and human assays over the past decades. As current pharmacological research is focused on anticancer and neurological action, it would be interesting and important to study the main compounds present in Aloe vera for therapeutic purposes.
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Objective:To screen small-molecule inhibitors of tyrosine kinase receptor B2 (EphB2) by using a molecular docking method, and to investigate their effect on cutaneous squamous cell carcinoma (CSCC) and possible mechanisms of action.Methods:The three-dimensional structure of EphB2 protein and its ligand binding sites were predicted by using the docking tool Schrodinger, and high-throughput virtual screening of EphB2 inhibitors was carried out by molecular docking. The anti-CSCC effect and mechanism of action of the screened EphB2 inhibitors kaempferitrin and aloe-emodin (AE) were verified in in vitro and in vivo experiments. In the in vitro experiments, human CSCC cell lines A431 and SCL-1, as well as the human immortalized keratinocyte HaCaT, were all divided into blank control group, dimethyl sulfoxide (DMSO) group, AE group and kaempferitrin group. Methyl thiazol tetrazolium (MTT) assay (AE at concentrations of 20, 40, 80, 160 μmol/L, kaempferitrin at concentrations of 12.5, 25, 50, 100 μmol/L), scratch and Transwell assays (AE at a fixed concentration of 80 μmol/L, kaempferitrin at a fixed concentration of 50 μmol/L) were performed to analyze the effect of EphB2 inhibitors on the proliferation, migration and invasion of CSCC cells. In the in vivo experiments, specific pathogen-free BALB/c female nude mice were subcutaneously injected with 0.2 ml of A431 cell suspension. After tumor growth, 24 tumor-bearing mice were randomly and equally divided into 4 groups: AE group and kaempferitrin group intraperitoneally injected with 20 mg·kg -1·d -1 AE and 25 mg·kg -1·d -1 kaempferitrin respectively, blank control group and DMSO group intraperitoneally injected with the same volume of sodium chloride physiological solution and DMSO respectively; the tumor size and body weight of nude mice were measured weekly; after consecutive treatment for 28 days, transplanted tumors were resected from the nude mice for hematoxylin and eosin (HE) staining, and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to analyze the effect of AE and kaempferitrin on the mRNA and protein expression of E-cadherin, vimentin, glycogen synthase kinase 3β (GSK-3β), phosphorylated GSK-3β (p-GSK-3β) and β-catenin respectively. One-way analysis of variance and t test were used for comparisons between groups. Results:Two small-molecule compounds AA-504/20999031 (kaempferitrin) and AA-466/21162055 (AE) with high inhibitory activity against EphB2 were screened out. MTT assay showed that both AE and kaempferitrin exhibited strong cytotoxicity to SCL-1 and A431 cells compared with HaCaT cells, and their toxicity increased with the increase of their concentration ( F = 17.95, 11.34, respectively, both P < 0.001) ; after 48-hour treatment, the 50% inhibitory concentrations (IC50s) of AE against SCL-1 and A431 cells were 124.59 and 80.85 μmol/L respectively, and the IC50s of kaempferitrin against SCL-1 and A431 cells were 119.64 and 64.96 μmol/L respectively. Scratch assay showed that the migration distance of A431 cells was significantly shorter in the AE group and kaempferitrin group (36.7 ± 1.0 μm, 44.7 ± 3.5 μm, respectively) than in the DMSO group (88.1 ± 1.4 μm, F = 52.34, P < 0.001), while there was no significant difference in the migration distance of HaCaT cells among the above groups ( F = 1.73, P = 0.238). Transwell assay showed that the number of A431 cells crossing the Transwell membrane significantly decreased in the AE group and kaempferitrin group (145.0 ± 2.5, 94.7 ± 4.1, respectively) compared with the DMSO group (195.3 ± 5.7, F = 72.85, P < 0.001), while neither AE nor kaempferitrin showed significant inhibitory effects of on the number of HaCaT cells crossing the Transwell membrane ( F = 3.91, P = 0.055). The animal experiment revealed significantly decreased volumes of transplanted tumors in nude mice in the AE group and kaempferitrin group (407.42 ± 70.37 mm 3, 368.77 ± 62.7 mm 3, respectively) compared with the DMSO group (841.88 ± 84.63 mm 3, F = 73.78, P < 0.001). HE staining confirmed that AE and kaempferitrin could improve pathological changes of transplanted tumors. qRT-PCR and Western blot analysis showed that AE and kaempferitrin significantly up-regulated the mRNA and protein expression of E-cadherin and p-GSK-3β in tumor tissues (all P < 0.001), and down-regulated the mRNA and protein expression of vimentin, β-catenin and GSK-3β (all P < 0.001) . Conclusion:The small-molecule inhibitors screened by molecular docking can form a stable complex with EphB2, and inhibit the progression of CSCC by affecting the Wnt/β-catenin pathway-induced epithelial-mesenchymal transition.
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ObjectiveTo study the inhibitory effect of aloe-emodin （AE） on aluminum ion （Al3+）-induced β-amyloid protein 42 （Aβ42） aggregation and its depolymerization on formed Aβ42-Al3+ aggregates in vitro， and to investigate the effect of AE on the cytotoxicity of Aβ42 aggregation in the presence of Al3+. MethodThe Aβ42 group， Aβ42+Al3+ group， Aβ42+AE group， Aβ42+Al3++AE group and the depolymerization test group were set up in the experiment. The aggregation fibrosis process， aggregation morphology， aggregation size and cytotoxicity of Aβ42 in each group were detected by thioflavin T （ThT） fluorescence assay， transmission electron microscopy （TEM）， dynamic light scattering （DLS） experiment and thiazolyl blue （MTT） cytotoxicity assay. ResultCompared with the Aβ42 group， Al3+ could promote Aβ42 aggregation， increase the fluorescence intensity of ThT by 124.48%， induce the aggregation of Aβ42 to form fiber bundles with larger particle size， and significantly reduce the cell viability of human neuroblastoma SH-SY5Y cells （P<0.01）， thus reducing the cell survival rate to 51.05%. AE not only inhibited Aβ42 aggregation， but also inhibited Al3+-induced Aβ42 aggregation in a concentration-dependent manner. Compared with the Aβ42+Al3+ group， high concentration of AE could reduce the ThT fluorescence intensity to 41.66%， and change the polypeptide aggregation pathway to form amorphous aggregates with small particle size. Besides， it significantly inhibited the cytotoxicity of Aβ42 induced by Al3+ （P<0.01）， and restored the cell survival rate to 84.87%. Further depolymerization was conducted， AE could depolymerize Aβ42-Al3+ aggregates to make the formed aggregates disappear and form some small-particle short fibers and amorphous structure aggregates with low toxicity. ConclusionAE can inhibit Aβ42 aggregation and cytotoxicity in the presence of Al3+， depolymerize the formed Aβ42-Al3+ aggregates and alleviate the cytotoxicity， thus laying the foundation for exploring the mechanism of AE in the treatment of Alzheimer's disease.
الملخص
ObjectiveTo study the inhibitory effect of aloe-emodin （AE） on aluminum ion （Al3+）-induced β-amyloid protein 42 （Aβ42） aggregation and its depolymerization on formed Aβ42-Al3+ aggregates in vitro， and to investigate the effect of AE on the cytotoxicity of Aβ42 aggregation in the presence of Al3+. MethodThe Aβ42 group， Aβ42+Al3+ group， Aβ42+AE group， Aβ42+Al3++AE group and the depolymerization test group were set up in the experiment. The aggregation fibrosis process， aggregation morphology， aggregation size and cytotoxicity of Aβ42 in each group were detected by thioflavin T （ThT） fluorescence assay， transmission electron microscopy （TEM）， dynamic light scattering （DLS） experiment and thiazolyl blue （MTT） cytotoxicity assay. ResultCompared with the Aβ42 group， Al3+ could promote Aβ42 aggregation， increase the fluorescence intensity of ThT by 124.48%， induce the aggregation of Aβ42 to form fiber bundles with larger particle size， and significantly reduce the cell viability of human neuroblastoma SH-SY5Y cells （P<0.01）， thus reducing the cell survival rate to 51.05%. AE not only inhibited Aβ42 aggregation， but also inhibited Al3+-induced Aβ42 aggregation in a concentration-dependent manner. Compared with the Aβ42+Al3+ group， high concentration of AE could reduce the ThT fluorescence intensity to 41.66%， and change the polypeptide aggregation pathway to form amorphous aggregates with small particle size. Besides， it significantly inhibited the cytotoxicity of Aβ42 induced by Al3+ （P<0.01）， and restored the cell survival rate to 84.87%. Further depolymerization was conducted， AE could depolymerize Aβ42-Al3+ aggregates to make the formed aggregates disappear and form some small-particle short fibers and amorphous structure aggregates with low toxicity. ConclusionAE can inhibit Aβ42 aggregation and cytotoxicity in the presence of Al3+， depolymerize the formed Aβ42-Al3+ aggregates and alleviate the cytotoxicity， thus laying the foundation for exploring the mechanism of AE in the treatment of Alzheimer's disease.
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Emodin nanostructured lipid carriers(ED-NLC) were prepared and their quality was evaluated in vitro. Based on the results of single-factor experiments, the ED-NLC formulation was optimized by Box-Behnken response surface method with the dosages of emodin, isopropyl myristate and poloxamer 188 as factors and the nanoparticle size, encapsulation efficiency and drug loading as evaluation indexes. Then the evaluation was performed on the morphology, size and in vitro release of the nanoparticles prepared by emulsification-ultrasonic dispersion method in line with the optimal formulation, i.e., 3.27 mg emodin, 148.68 mg isopropyl myristate and 173.48 mg poloxamer 188. Under a transmission electron microscope(TEM), ED-NLC were spherical and their particle size distribution was uniform. The particle size of ED-NLC was(97.02±1.55) nm, the polymer dispersion index 0.21±0.01, the zeta potential(-38.96±0.65) mV, the encapsulation efficiency 90.41%±0.56% and the drug loading 1.55%±0.01%. The results of differential scanning calorimeter(DSC) indicated that emodin may be encapsulated into the nanostructured lipid carriers in molecular or amorphous form. In vitro drug release had obvious characteristics of slow release, which accorded with the first-order drug release equation. The fitting model of Box-Behnken response surface methodology was proved accurate and reliable. The optimal formulation-based ED-NLC featured concentrated particle size distribution and high encapsulation efficiency, which laid a foundation for the follow-up study of ED-NLC in vivo.
الموضوعات
Drug Carriers , Emodin , Follow-Up Studies , Lipids , Nanostructuresالملخص
The present study investigated the effect of emodin on the serum metabolite profiles in the chronic constriction injury(CCI) model by non-target metabolomics and explored its analgesic mechanism. Twenty-four Sprague Dawley(SD) rats were randomly divided into a sham group(S), a CCI group(C), and an emodin group(E). The rats in the emodin group were taken emodin via gavage once a day for fifteen days(50 mg·kg~(-1)) on the first day after the CCI surgery. Mechanical withdrawal threshold(MWT) and thermal withdrawal threshold(TWL) in each group were performed before the CCI surgery and 3,7, 11, and 15 days after surgery. After 15 days, blood samples were collected from the abdominal aorta. The differential metabolites were screened out by non-target metabolomics and analyzed with Kyoto Encyclopedia of Genes and Genomes(KEGG) and ingenuity pathway analysis(IPA). From the third day after CCI surgery, the MWT and TWL values were reduced significantly in both CCI group and emodin group, compared with the sham group(P<0.01). At 15 days post-surgery, the MWT and TWL values in emodin group increased significantly compared with the CCI group(P<0.05). As revealed by non-target metabolomics, 72 differential serum metabolites were screened out from the C-S comparison, including 41 up-regulated and 31 down-regulated ones, while 26 differential serum metabolites from E-C comparison, including 10 up-regulated and 16 down-regulated ones. KEGG analysis showed that the differential metabolites in E-C comparison were enriched in the signaling pathways, such as sphingolipid metabolism, arginine biosynthesis, glycerophospholipid metabolism, and tryptophan metabolism. IPA showed that the differential metabolites were mainly involved in the lipid metabolism-molecular transport-small molecule biochemistry network. In conclusion, emodin can exert an analgesic role via regulating sphingolipid metabolism and arginine biosynthesis.