Effects of Tilianin on Proliferation,Migration and TGF‐β/Smad Signaling in Rat Vascular Smooth Muscle Cells Induced with Angiotensin II

Flavonoid Tilianin was isolated from Dracocephalum moldavica, and its pharmacological mechanism on proliferation, migration and the TGF‐β/Smad signaling pathway in rat vascular smooth muscle cells (VSMCs) induced with Angiotensin II (Ang II) was systematically evaluated. Primary rat VSMCs were stimulated with Ang II to induce proliferation. The cells were then treated with Tilianin for 24 or 48 h. MTT assay and Transwell assays were used to evaluate the effects of Tilianin on proliferation and migration. The expression of intracellular proliferating cell nuclear antigen (PCNA), intercellular adhesion molecule‐1 (ICAM‐1) and vascular cell adhesion molecule‐1 (VCAM‐1) were measured by immunohistochemistry as verification of effects on proliferation and migration. The expression of TGF‐β1, Smad2 and Smad3 mRNA was measured by qRT‐PCR, and the expression of TGF‐β1 and P‐Smad2/3 protein was measured by Western blotting. The results show that Tilianin can inhibit proliferation and expression of intracellular PCNA in VSMCs induced with Ang II, in a dose‐dependent manner. Tilianin also mediates a dose‐dependent inhibition of migration and the expression of intracellular ICAM‐1, VCAM‐1, MMP‐2 and MMP‐9. Furthermore, TGF‐β1, Smad2, Smad3, Smad2/3 and P‐Smad2/3 in Ang II‐induced VSMCs are suppressed by Tilianin. The inhibitory effects of Tilianin support its use in the suppression and treatment of atherosclerosis. Copyright © 2017 John Wiley & Sons, Ltd.     

补益营卫方对TGFβ1/Smads信号转导途径基因表达的影响

目的:研究补益营卫方对衰老人皮肤成纤维细胞TGFβ1/Smads信号蛋白基因表达的影响。方法:通过传代培养复制衰老人皮肤成纤维细胞模型,利用荧光定量PCR法检测补益营卫方对衰老人皮肤成纤维细胞TGFβ1/Smads信号蛋白基因表达。结果:人皮肤成纤维细胞衰老时Smad2基因的表达水平,与年轻细胞相比,变化趋势基本一致。补益营卫药物处理后Smad2基因的表达,24h和72h时,分别是衰老人皮肤成纤维细胞基因相对表达量的2.85倍和2.5倍;而48 h时则两者相当。衰老时Smad3基因的表达水平,24 h和48 h时,衰老人皮肤成纤维细胞比年轻细胞分别下降了29%和36%;72 h则增加了3.95倍。补益营卫药物处理后Smad3基因的表达,24 h、48 h和72 h时,分别是衰老人皮肤成纤维细胞的3.15倍、2.6倍和1.6倍。衰老时Smad4基因的表达水平,24 h时比年轻细胞下降了32%,但是,48 h和72 h时则分别是年轻人皮肤成纤维细胞的1.6倍和约3倍。补益营卫药物处理后Smad4基因的表达24 h、48 h和72 h分别是衰老组的2.8倍、1.6倍和2倍。衰老时Smad7基因的表达水平,24 h和48 h时分别比年轻细胞下降了88%和64%。但是,72 h时则达到年轻人皮肤成纤维细胞表达水平的1.9倍。补益营卫药物处理后Smad7基因的表达,其24h时是衰老人皮肤成纤维细胞表达水平的4.24倍,但是,48 h和72 h则分别下降了24%和33%。结论:衰老时TGFβ1/Smads信号蛋白表达紊乱;补益营卫方对衰老人皮肤成纤维细胞Smad2、Smad3、Smad4基因表达有促进作用;对Smad7基因表达表现为双向调节作用。     

Hydroxysafflor Yellow A Attenuates Bleomycin‐induced Pulmonary Fibrosis in Mice

Hydroxysafflor yellow A (HSYA) is an active component of Carthamus tinctorius L., and we want to investigate whether HSYA attenuates pulmonary fibrosis induced by bleomycin (BLM) in mice. The mice received a BLM via oropharyngeal aspiration, and HSYA was intraperitoneally injected. Arterial blood gas analysis was performed. Morphological changes and hydroxyproline content were measured. mRNA expression of transforming growth factor‐β1 (TGF‐β1), connective tissue growth factor, α‐smooth muscle actin (α‐SMA), and collagen I was measured by real‐time polymerase chain reaction. α‐SMA‐positive cells in lung tissues were detected by immunohistochemical staining. A549 cell was cultured, and morphological changes were observed after TGF‐β1 and HSYA treatment. mRNA expression was detected by real‐time polymerase chain reaction. Phosphorylation of Smad3 was evaluated by western blotting. HSYA decreased the lung consolidation area and collagen deposition in mice with pulmonary fibrosis. The blood gas changes due to BLM were attenuated by HSYA. HSYA also alleviated the BLM‐induced increase of TGF‐β1, connective tissue growth factor, α‐SMA, and collagen I mRNA levels. HSYA treatment inhibited the increase of α‐SMA expression, Smad3 phosphorylation, the morphological changes in lung tissue. HSYA inhibits Smad3 phosphorylation and elevated expression of collagen I mRNA in epithelial–mesenchymal transition induced by TGF‐β1. Copyright © 2016 John Wiley & Sons, Ltd.     

(+)‐Isobicyclogermacrenal and spathulenol from Aristolochia yunnanensis alleviate cardiac fibrosis by inhibiting transforming growth factor β/small mother against decapentaplegic signaling pathway

Cardiac fibrosis contributes to both systolic and diastolic dysfunction in many cardiac pathophysiologic conditions. Antifibrotic therapies are likely to be a crucial strategy in curbing many fibrosis‐related cardiac diseases. In our previous study, an ethyl acetate extract of a traditional Chinese medicine Aristolochia yunnanensis Franch. was found to have a therapeutic effect on myocardial fibrosis in vitro and in vivo. However, the exact chemicals and their mechanisms responsible for the activity of the crude extract have not been illustrated yet. In the current study, 10 sesquiterpenoids ( 1 – 10 ) were isolated from the active extract, and their antifibrotic effects were systematically evaluated in transforming growth factor β 1 (TGFβ1)‐stimulated cardiac fibroblasts and NIH3T3 fibrosis models. (+)‐Isobicyclogermacrenal ( 1 ) and spathulenol ( 2 ) were identified as the main active components, being more potent than the well‐known natural antifibrotic agent oxymatrine. Compounds 1 and 2 could inhibit the TGFβ1‐induced cardiac fibroblasts proliferation and suppress the expression of the fibrosis biomarkers fibronectin and α‐smooth muscle actin via down‐regulation of their mRNA levels. The mechanism study revealed that 1 and 2 could inhibit the phosphorylation of TGFβ type I receptor, leading to the decrease of the phosphorylation levels of downstream Smad2/3, then consequently blocking the nuclear translocation of Smad2/3 in the TGFβ/Smad signaling pathway. These findings suggest that 1 and 2 may serve as promising natural leads for the development of anticardiac fibrosis drugs.     

Corosolic Acid Exhibits Anti‐angiogenic and Anti‐lymphangiogenic Effects on In Vitro Endothelial Cells and on an In Vivo CT‐26 Colon Carcinoma Animal Model

We describe the anti‐angiogenic and anti‐lymphangiogenic effects of corosolic acid, a pentacyclic triterpenoid isolated from Cornus kousa Burg. A mouse colon carcinoma CT‐26 animal model was employed to determine the in vivo anti‐angiogenic and anti‐lymphangiogenic effects of corosolic acid. Corosolic acid induced apoptosis in CT‐26 cells, mediated by the activation of caspase‐3. In addition, it reduced the final tumor volume and the blood and lymphatic vessel densities of tumors, indicating that it suppresses in vivo angiogenesis and lymphangiogenesis. Corosolic acid inhibited the proliferation and tube formation of human umbilical vein endothelial cells and human dermal lymphatic microvascular endothelial cells. In addition, corosolic acid decreased the proliferation and migration of human umbilical vein endothelial cells stimulated by angiopoietin‐1. Pretreatment with corosolic acid decreased the phosphorylation of focal adhesion kinase (FAK) and ERK1/2, suggesting that corosolic acid contains anti‐angiogenic activity that can suppress FAK signaling induced by angiopoietin‐1. Copyright © 2015 John Wiley & Sons, Ltd.     

Astragaloside IV inhibits TGF‐β1‐induced epithelial‐mesenchymal transition through inhibition of the PI3K/Akt/NF‐κB pathway in gastric cancer cells

Astragaloside IV (AS‐IV) has been reported to possess anti‐metastasis activity in cancer cells. However, it is unknown whether AS‐IV could inhibit epithelial‐mesenchymal transition (EMT), a cellular de‐differentiation program that promotes metastasis, in cancer cells. The aim of this study was to study the effect and mechanism of AS‐IV on EMT in gastric cancer (GC) cells. The results showed that AS‐IV significantly inhibited cell viability, invasion, and migration of GC cells. The E‐cadherin to N‐cadherin switch and expression of Vimentin and metastasis‐related genes were induced by transforming growth factor β1 (TGF‐β1), whereas AS‐IV reversed the induction. In addition, AS‐IV inhibited TGF‐β1‐induced activation of PI3K/Akt/NF‐κB. Inhibition of the PI3K/Akt/NF‐κB pathway reversed TGF‐β1‐induced EMT. In conclusion, AS‐IV inhibited TGF‐β1‐induced EMT through inhibition of the PI3K/Akt/NF‐κB pathway in GC cells. AS‐IV might be an effective candidate for the treatment for GC.     

Camphor Induces Proliferative and Anti‐senescence Activities in Human Primary Dermal Fibroblasts and Inhibits UV‐Induced Wrinkle Formation in Mouse Skin

Camphor ((1R)‐1,7,7‐trimethylbicyclo[2.2.1]heptan‐2‐one), a bicyclic monoterpene, is one of the major constituents of essential oils from various herbs such as rosemary, lavender, and sage. In this study, we investigated the beneficial effects of camphor as a botanical ingredient in cosmetics. Camphor induced the proliferation of human primary dermal fibroblasts in a dose‐dependent manner via the PI3K/AKT and ERK signaling pathways. Camphor attenuated the elevation of senescence associated with β‐galactosidase (SA‐β‐gal) activity. Elastase activity decreased, while the total amount of collagen increased, in a dose‐ and time‐dependent manner in human primary dermal fibroblasts treated with camphor. Camphor induced the expression of collagen IA, collagen IIIA, collagen IVA, and elastin in human primary dermal fibroblasts. In addition, posttreatment with 26 and 52 mM camphor for 2 weeks led to a significant reduction in the expression of MMP1 but increases in the expression of collagen IA, IIIA, and elastin in mouse skin exposed to UV for 4 weeks. These posttreatments also reduced the depths of the epidermis and subcutaneous fat layer in UV‐exposed mouse skin. Taken together, these findings suggest camphor to be a potent wound healing and antiwrinkle agent with considerable potential for use in cosmeceuticals. Copyright © 2015 John Wiley & Sons, Ltd.     

Coriolus versicolor alleviates diabetic cardiomyopathy by inhibiting cardiac fibrosis and NLRP3 inflammasome activation

Coriolus versicolor (CV) is a traditional medicine and food mushroom. Our previous study demonstrated that CV extract exhibited anti‐hyperglycemia and anti‐insulin resistance effects. However, the effect of CV on cardiac function in diabetic cardiomyopathy (DCM) remains unclear. Therefore, we aimed to investigate the effect of CV on cardiac function in diabetes mellitus (DM) rats. We found that the cardiac dysfunction of DM rats was markedly improved by CV extract treatment. CV extract administration significantly attenuated cardiac fibrosis in DM rats, which was accompanied by suppressed transforming growth factor beta 1 (TGF‐β1)/Smad signaling as indicated by decreased levels of TGF‐β1, p‐Smad2, and p‐Smad3 and increased Smad7 expression. Moreover, CV extract treatment significantly alleviated cardiac inflammation as shown by decreased levels of NLRP3 receptor, cleaved caspase‐1, IL‐1β, and IL‐18 in DM rats at least partly due to the inhibition of the NF‐κB. In addition, high‐glucose treatment induced cardiac fibrosis and increased cardiac inflammation in cardiac fibroblast cells, but these effects were ameliorated by CV extract treatment. Therefore, we conclude that the protective effect of CV on DCM is associated with the suppression of TGF‐β1/Smad signaling and attenuation of NLRP3 inflammasome activation, suggesting that CV extract may be a potential therapeutic agent for DCM.     

Madecassoside Ameliorates Bleomycin‐Induced Pulmonary Fibrosis in Mice by Downregulating Collagen Deposition

This study aimed to explore the protective effects of madecassoside (Mad), a triterpenoid saponin isolated from Centella asiatica herbs, on experimental pulmonary fibrosis (PF) and underlying mechanisms. PF model was established in mice by endotracheal instillation with bleomycin (5 mg/kg). Mice were orally administered with Mad (10, 20, 40 mg/kg) and prednisone (5 mg/kg) for 7 or 21 days. Mad (20, 40 mg/kg) significantly improved lung pathological changes and reduced collagen deposition. In the aspect of collagen synthesis, Mad (20, 40 mg/kg) reduced the expressions of α‐smooth muscle actin and transforming growth factor‐β1 (TGF‐β1), and inhibited the phosphorylations of Smad2 and Smad3 in the lung tissues. However, in vitro, Mad showed little effect on TGF‐β1‐induced phosphorylation of either Smad2 or Smad3 in primary mouse lung fibroblasts. Moreover, Mad (20, 40 mg/kg) attenuated oxidative damage and inflammation presented at the early stage of PF, evidenced by reduced total leukocytes in the bronchoalveolar lavage fluid, decreased myeloperoxidase activity and malondialdehyde level, and increased super‐oxide dismutase activity and glutathione level in lung tissues. On the other hand, Mad (40 mg/kg) elevated the matrix metalloproteinase 1/tissue inhibitor of metalloproteinase 1 ratio in lung tissues of PF mice mainly by downregulating tissue inhibitor of metalloproteinase 1 expression. The present study demonstrated that Mad can ameliorate PF by preventing the deposition of extracellular matrix, which might be achieved mainly through attenuating inflammation and oxidative stress and consequent TGF‐β1 overexpression. Copyright © 2014 John Wiley & Sons, Ltd.     

A Comparative Study of Baby Immature and Adult Shoots of Aloe Vera on UVB‐Induced Skin Photoaging in vitro

Ultraviolet (UV) irradiation induces photo‐damage of the skin, which in turn causes depletion of the dermal extracellular matrix and chronic alterations in skin structure. Skin wrinkle formations are associated with collagen synthesis and matrix metalloproteinase (MMP) expression. The production of type I procollagen is regulated by transforming growth factor‐β1 (TGF‐β1) expression; the activation of MMP is also correlated with an increase of interleukin‐6 (IL‐6). Aloe barbadensis M. (Aloe vera) is widely used in cosmetic and pharmaceutical products. In this study, we examined whether baby aloe shoot extract (BAE, immature aloe extract), which is from the one‐month‐old shoots of Aloe vera, and adult aloe shoot extract (AE), which is from the four‐month‐old shoots of Aloe vera, have a protective effect on UVB‐induced skin photoaging in normal human dermal fibroblasts (NHDFs). The effects of BAE and AE on UVB‐induced photoaging were tested by measuring the levels of reactive oxygen species, MMP‐1, MMP‐3, IL‐6, type I procollagen, and TGF‐β1 after UVB irradiation. We found that NHDF cells treated with BAE after UVB‐irradiation suppressed MMP‐1, MMP‐3, and IL‐6 levels compared to the AE‐treated cells. Furthermore, BAE treatment elevated type I procollagen and TGF‐β1 levels. Our results suggest that BAE may potentially protect the skin from UVB‐induced damage more than AE. Copyright © 2013 John Wiley & Sons, Ltd.     

Schisandrin B attenuates epidural fibrosis in postlaminectomy rats by inhibiting proliferation and extracellular matrix production of fibroblasts

Laminectomy has been widely considered one of the most common treatments for lumbar disorders. Epidural fibrosis (EF) is a common complication after laminectomy, causing recurrent postoperative pain. Schisandrin B (Sch.B), the active ingredient extracted from Schisandra chinensis Fructus, has been found to have potent antiproliferative and antifibrotic effects on several cells. This study aimed to investigate the effects of Sch.B on the prevention of postlaminectomy EF formation. In vitro, we studied the effects of Sch.B on transforming growth factor beta 1 (TGF‐β1)‐induced proliferation and extracellular matrix (ECM) production of primary fibroblasts, as well as its underlying mechanism. We found that Sch.B not only inhibited the proliferation of fibroblasts but also reduced ECM production, including that of connective tissue growth factor, fibronectin, and type I collagen, in a dose‐dependent manner. Mechanistically, we found that Sch.B suppressed TGF‐β1‐stimulated activation of the Smad2/3 and mitogen‐activated protein kinase pathways. Moreover, the in vivo study demonstrated that Sch.B treatment attenuated the progression of EF in a postlaminectomy rat model via reducing the cell number and ECM production of scar tissue. Taken together, these data suggested that Sch.B possesses great potential value as a preventative agent for EF.     

Labda‐8(17),12,14‐trien‐19‐oic Acid Contained in Fruits of Cupressus sempervirens Suppresses Benign Prostatic Hyperplasia in Rat and In Vitro Human Models Through Inhibition of Androgen and STAT‐3 Signaling

Fruit extract of Cupressus sempervirens (CS), which is used traditionally to treat Benign Prostatic Hyperplasia (BPH)‐like urinary symptoms in patients, was scientifically validated for anti‐BPH activity. The ethanolic fruit extract of CS inhibited proliferation of human BPH‐stromal cells and the activity was localized to its chloroform‐soluble, diterpene‐rich fraction. Eight major diterpenes isolated from this fraction exhibited moderate to potent activity and the most active diterpene (labda‐8(17),12,14‐trien‐19‐oic acid) exhibited an IC₅₀ of 37.5 μM (antiproliferative activity against human BPH‐stromal cells). It significantly inhibited activation (phosphorylation) of Stat‐3 in BPH‐stromal cells and prevented transactivation of androgen sensitive KLK3/PSA and TMPRSS2 genes in LNCaP cells. Labda‐8(17),12,14‐trien‐19‐oic acid‐rich CS fraction prevented prostatic hyperplasia in rat model and caused TUNEL labeling of stromal cells with lower expressions of IGF‐I, TGF‐ß and PCNA, and bcl‐2/bax ratio. Human BPH tissues exhibited precise lowering of stromal component after incubation in labda‐8(17),12,14‐trien‐19‐oic acid, ex vivo. We conclude that labda‐8(17),12,14‐trien‐19‐oic acid contained in CS exhibits anti‐BPH activity through inhibition of stromal proliferation and suppression of androgen action in the prostate, presenting a unique lead structure for further optimization of anti‐BPH activity. Copyright © 2014 John Wiley & Sons, Ltd.     

Effects of Cyanidin‐3‐O‐glucoside on Synthetic and Metabolic Activity of Ethanol Stimulated Human Pancreatic Stellate Cells

Activated pancreatic stellate cells (PSC) play a major role in the development of chronic pancreatitis. Flavonoids (C‐3‐O‐G) theoretically may have potential to suppress activated PSC. The aim of our study was to determine the ability of C‐3‐O‐G to invert synthetic and metabolic activity of alcohol stimulated human pancreatic stellate cells (hPSC). In the present study we demonstrate that treatment with C‐3‐O‐G decreased proliferation rate of ethanol activated hPSC by 51%. Synthesis of extracellular matrix proteins in activated hPSC was markedly inhibited, as shown by reduced levels of collagen I and fibronectin expression. The decrease of secretion of fibronectin by 33% and in collagen I‐25% in ethanol activated and C‐3‐O‐G treated hPSC was observed. Moreover, treatment of ethanol activated hPSC with C‐3‐O‐G resulted in the decrease of oxygen consumption rate by 44% and reduced levels of ATP synthesis (i.e. energy production) by 41%. Hence, the effects of C‐3‐O‐G on ethanol activated hPSC may provide new insights for the use of anthocyanins as anti‐fibrogenic agents in treatment and/or prevention of pancreatic fibrosis. Copyright © 2015 John Wiley & Sons, Ltd.     

Diosmetin exhibits anti‐proliferative and anti‐inflammatory effects on TNF‐α‐stimulated human rheumatoid arthritis fibroblast‐like synoviocytes through regulating the Akt and NF‐κB signaling pathways

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by inflammation and proliferation of synovial tissues. Diosmetin is a bioflavonoid possessing an anti‐inflammatory property. Herein, we aimed to study the effects of diosmetin on the inflammation and proliferation of RA fibroblast‐like synoviocytes MH7A cells. MH7A cell proliferation was measured using cell counting kit‐8 assay. Cell apoptosis was examined using flow cytometry. The production of inflammatory cytokines including interleukin (IL)‐1β, IL‐6, IL‐8, and matrix metalloproteinase‐1 (MMP‐1) was measured using enzyme‐linked immunosorbent assay (ELISA). Results showed that diosmetin inhibited tumor necrosis factor‐α (TNF‐α)‐induced proliferation increase in MH7A cells in a dose‐dependent manner. Diosmetin treatment resulted in an increase in apoptotic rates and a reduction in TNF‐α‐induced production of IL‐1β, IL‐6, IL‐8, and MMP‐1 in MH7A cells. Furthermore, diosmetin inhibited TNF‐α‐induced activation of protein kinase B (Akt) and nuclear factor‐κB (NF‐κB) pathways in MH7A cells. Suppression of Akt or NF‐κB promoted apoptosis and inhibited TNF‐α‐induced proliferation increase and production of IL‐1β, IL‐6, IL‐8, and MMP‐1 in MH7A cells, and diosmetin treatment enhanced these effects. Taken together, these findings suggested that diosmetin exhibited anti‐proliferative and anti‐inflammatory effects via inhibiting the Akt and NF‐κB pathways in MH7A cells.     

The effect of xanthorrhizol on the expression of matrix metalloproteinase‐1 and type‐I procollagen in ultraviolet‐irradiated human skin fibroblasts

Matrix metalloproteinases (MMPs) are key regulators of the skin photoaging process that is set in motion by exposure to ultraviolet (UV) irradiation. This skin damage results from UV‐induced generation of reactive oxygen species, which are associated with upregulation of MMPs and decreased collagen synthesis. We investigated the effects of xanthorrhizol, isolated from Curcuma xanthorrhiza, on the expression of MMP‐1 and type‐I procollagen in UV‐irradiated human skin fibroblasts. Fibroblasts cultured in the presence or absence of purified xanthorrhizol or C. xanthorrhiza extract were irradiated with UV (20 mJ/cm²), and MMP‐1 and type‐I procollagen levels were measured using Western blot analysis. Xanthorrhizol (0.001–0.1 µM ) and C. xanthorrhiza extract (0.01–0.5 µg/mL) induced a significant, dose‐dependent decrease in the expression of MMP‐1 protein, and increased the expression of type‐1 procollagen. At a concentration of 0.1 µM , xanthorrhizol nearly completely abrogated MMP‐1 expression. The MMP‐1‐suppressing and type‐1 procollagen‐inducing effects of xanthorrhizol treatment were greater than those of epigallocatechin 3‐O‐gallate (EGCG), which is known to be a natural anti‐aging agent. These results suggest that xanthorrhizol is a potential candidate for the prevention and treatment of skin aging. Copyright © 2009 John Wiley & Sons, Ltd.     

Brassinin Combined with Capsaicin Enhances Apoptotic and Anti‐metastatic Effects in PC‐3 Human Prostate Cancer Cells

Brassinin (BSN), a type of indole compound derived from cruciferous vegetables, has shown anti‐cancer effects in cells and animals. Capsaicin (CAP), an alkaloid derived from the chilli pepper, is also of interest in for its reported efficacy against various malignancies. The objective of our study was to analyze the potential synergistic anti‐tumor effects of BSN combined with CAP on prostate cancer PC‐3 cells. After treatment with BSN and CAP at various concentrations, the synergistic cytotoxic effect of PC‐3 cells was analyzed by MTT method, proliferation, apoptosis, mitochondrial membrane potential, colony formation, and Western blotting. Moreover, the inhibitory effects of BSN and CAP on the constitutive expressions of MMP‐9/2, their enzymatic activities, cellular migration, and cell invasion were also investigated. The cytotoxicity was synergistically increased in combination compared with the single drug used; moreover, proliferation, apoptosis, mitochondrial membrane potential, and colony formation were significantly suppressed and anti‐apoptotic‐, proliferative‐, and metastatic‐related proteins were clearly abolished in the combination group. Besides, constitutive MMP‐9/2 expression, their enzymatic activities, cell migration, and tumor cell invasion were inhibited, and TIMP‐1 was up‐regulated in the combination group in PC‐3 cells. Our results indicate, for the first time, that BSN and CAP in combination exert synergistic anticancer effects in prostate carcinoma. Copyright © 2015 John Wiley & Sons, Ltd.     

Protective effects of rutin on rat glomerular mesangial cells cultured in high glucose conditions

The protective effect of rutin on the glomerulosclerosis of diabetic nephropathy (DN) in rat mesangial cells was investigated. The cultured mesangial cells were divided into eight groups: normal, solvent control, high glucose, low dose of rutin, moderate dose of rutin, high dose of rutin, captopril and Ginkgo biloba extract. The cell cycles, type IV collagen and laminin in cytoplasm, TGF‐β₁ mRNA of mesangial cells, Smad 2/3 and Smad 7, and the activities of four antioxidant indexes including T‐SOD, MDA, CAT and GSH‐Px were measured by flow cytometry, radioimmunoassay, RT‐PCR, western blotting and visible spectrophotometry, respectively. Compared with the high glucose group, rutin decreased the cell percentages of the G₀/G₁ phase and inhibited the expression of Smad 2/3, laminin and type IV collagen, and TGF‐β₁ mRNA level, significantly. The antioxidant capacity, the cell percentages of S phase and Smad 7 expression were significantly increased by rutin. These results suggest that rutin is a potent protective agent against glomerulosclerosis in DN. Copyright © 2011 John Wiley & Sons, Ltd.     

Zerumbone Suppresses IL‐1β‐induced Cell Migration and Invasion by Inhibiting IL‐8 and MMP‐3 Expression in Human Triple‐negative Breast Cancer Cells

Inflammation is a key regulatory process in cancer development. Prolonged exposure of breast tumor cells to inflammatory cytokines leads to epithelial‐mesenchymal transition, which is the principal mechanism involved in metastasis and tumor invasion. Interleukin (IL)‐1β is a major inflammatory cytokine in a variety of tumors. To date, the regulatory mechanism of IL‐1β‐induced cell migration and invasion has not been fully elucidated. Here, we investigated the effect of zerumbone (ZER) on IL‐1β‐induced cell migration and invasion in breast cancer cells. The levels of IL‐8 and matrix metalloproteinase (MMP)‐3 mRNA were analyzed by real‐time polymerase chain reaction. The levels of secreted IL‐8 and MMP‐3 protein were analyzed by enzyme‐linked immunosorbent assay and western blot analysis, respectively. Cell invasion and migration was detected by Boyden chamber assay. The levels of IL‐8 and MMP‐3 expression were significantly increased by IL‐1β treatment in Hs578T and MDA‐MB231 cells. On the other hand, IL‐1β‐induced IL‐8 and MMP‐3 expression was decreased by ZER. Finally, IL‐1β‐induced cell migration and invasion were decreased by ZER in Hs578T and MDA‐MB231 cells. ZER suppresses IL‐1β‐induced cell migration and invasion by inhibiting IL‐8 expression and MMP‐3 expression in TNBC cells. ZER could be a promising therapeutic drug for treatment of triple‐negative breast cancer patients. Copyright © 2014 John Wiley & Sons, Ltd.     

HIF‐1α/VEGF signaling‐mediated epithelial–mesenchymal transition and angiogenesis is critically involved in anti‐metastasis effect of luteolin in melanoma cells

Tumor metastasis is still the leading cause of melanoma mortality. Luteolin, a natural flavonoid, is found in fruits, vegetables, and medicinal herbs. The pharmacological action and mechanism of luteolin on the metastasis of melanoma remain elusive. In this study, we investigated the effect of luteolin on A375 and B16‐F10 cell viability, migration, invasion, adhesion, and tube formation of human umbilical vein endothelial cells. Epithelial–mesenchymal transition (EMT) markers and pivotal molecules in HIF‐1α/VEGF signaling expression were analysed using western blot assays or quantitative real‐time polymerase chain reaction. Results showed that luteolin inhibits cellular proliferation in A375 and B16‐F10 melanoma cells in a time‐dependent and concentration‐dependent manner. Luteolin significantly inhibited the migratory, invasive, adhesive, and tube‐forming potential of highly metastatic A375 and B16‐F10 melanoma cells or human umbilical vein endothelial cells at sub‐IC₅₀ concentrations, where no significant cytotoxicity was observed. Luteolin effectively suppressed EMT by increased E‐cadherin and decreased N‐cadherin and vimentin expression both in mRNA and protein levels. Further, luteolin exerted its anti‐metastasis activity through decreasing the p‐Akt, HIF‐1α, VEGF‐A, p‐VEGFR‐2, MMP‐2, and MMP‐9 proteins expression. Overall, our findings first time suggests that HIF‐1α/VEGF signaling‐mediated EMT and angiogenesis is critically involved in anti‐metastasis effect of luteolin as a potential therapeutic candidate for melanoma.