橙皮苷对TGF-β1诱导的A549非小细胞肺癌细胞上皮间质转化的影响

目的：探讨橙皮苷对转化生长因子-β1（transforming growth factor-β1,TGF-β1）诱导的人非小细胞肺癌细胞上皮-间质转化的影响。方法：以A549人非小细胞肺癌细胞系作为研究对象。用MTT法,分别检测橙皮苷对正常培养条件或TGF-β1刺激下细胞增殖的影响。应用2种方法评估橙皮苷对TGF-β1刺激条件下A549细胞的上皮间质转化：圆形度值定量评估细胞的形态变化;双抗体夹心酶联免疫吸附法（enzyme linked-immuno-sorbent assay,ELISA）检测上皮细胞钙黏蛋白（E-cadherin,E-cad）、ɑ-平滑肌肌动蛋白（ɑ-smooth muscle actin,ɑ-SMA）、基质金属蛋白酶抑制因子-1（tissue inhibitors of metalloproteinases-1,TIMP-1）和基质金属蛋白酶-9（metallopreoteinases-9,MMP-9）水平。结果：0~40 μmol·L^-1的橙皮苷对正常培养条件或TGF-β1刺激后A549细胞的增殖没有显著影响。5 ng·mL^-1 TGF-β1能够诱导A549细胞的上皮间质转化：由上皮特征的铺路石状的细胞形态转化为梭形形态,圆形度值减少;E-cad分泌量减少和ɑ-SMA合成量增加。另外,MMP-9分泌量增加,MMP-9/TIMP-1比值增加。在细胞出现形态改变后,应用橙皮苷（20 μmol·L^-1或40 μmol·L^-1）能够降低ɑ-SMA、MMP-9水平和MMP-9/TIMP-1比值,增加E-cad水平,对细胞的圆形度值没有影响。结论：橙皮苷减轻TGF-β1诱导的非小细胞肺癌细胞的上皮间质转化程度,对非小细胞肺癌的转移和扩散有一定的抑制作用。     

MAPK pathway mediates epithelial‐mesenchymal transition induced by paraquat in alveolar epithelial cells

Epithelial‐mesenchymal transition (EMT) is believed to be involved in lung fibrosis process induced by paraquat (PQ); however, the molecular mechanism of this process has not been clearly established. The present study investigated the potential involvement of EMT after PQ poisoning. The expressions of EMT markers, such as E‐cadherin and α‐smooth muscle actin (α‐SMA), at multiple time points after exposure to different concentrations of PQ were evaluated by western blot analysis. Following PQ treatment, EMT induction was observed under microscopy. Related fibrosis genes, including Matrix metalloproteinase 2 (MMP‐2), Matrix metalloproteinase 9 (MMP‐9), collagens type I (COL I), and type III (COL III), were also evaluated by measuring their mRNA levels using RT‐PCR analysis. Signaling pathways were analyzed using selective pharmacological inhibitors for MAPK. Cell migration ability was evaluated by scratch wound and Transwell assays. The data showed that PQ‐induced epithelial RLE‐6NT cells to develop mesenchymal cell characteristics, as indicated by a significant decrease in the epithelial marker E‐cadherin and a significant increase in the extracellular matrix (ECM) marker α‐smooth muscle actin in a dose and time‐dependent manner. Moreover, PQ‐treated RLE‐6NT cells had an EMT‐like phenotype with elevated expression of MMP‐2, MMP‐9, and COL I and COL III and enhanced migration ability. Signal pathway analysis revealed that PQ‐induced EMT led to ERK‐1 and Smad2 phosphorylation through activation of the MAPK pathway. The results of the current study indicate that PQ‐induced pulmonary fibrosis occurs via EMT, which is mediated by the MAPK pathway. This implies that the MAPK pathway is a promising therapeutic target in alveolar epithelial cells. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1407–1414, 2016.     

内皮素-1活化ROCK/SLUG诱导人卵巢癌细胞发生上皮向间充质转分化

目的研究ROCK/SLUG信号通路在内皮素-1(endo-thelin-1,ET-1)促进人卵巢癌细胞上皮向间充质转分化(epi-thelial to mesenchymal transition,EMT)中的作用。方法 ET-1处理人卵巢癌细胞株SK-OV-3和CaOV3,或共用ROCK的活化突变体转染细胞或加入ROCK的抑制剂Y27632,并转染含SLUG启动子的pGL3质粒与Renilla质粒。实验末用Boyden小室体外侵袭实验检测细胞侵袭能力,细胞免疫荧光染色观察细胞形态,报告基因检测试剂盒检测SLUG启动子活性,用实时定量PCR和Western bot方法检测EMT相关基因的表达。结果 ET-1诱导SK-OV-3和CaOV3发生与EMT相一致的形态和基因变化,促进其细胞侵袭力;ET-1与内皮素A受体(endothelin A receptor,ETAR)结合,促进转录因子SLUG的转录;ET-1促进ROCK及fibronectin的表达,同时转染ROCK的活化突变体,促进ET-1诱导的fibronectin表达以及细胞侵袭力的增加。相反,ROCK抑制剂Y27632抑制ET-1对fibronectin表达以及细胞侵袭力的促进作用;转染ROCK的活化突变体,上调SLUG基因转录启动子活性促进其转录,抑制E-cadherin的转录。相反,ROCK的抑制剂Y27632抑制SLUG基因启动子的活性。结论 ET-1通过活化ROCK/SLUG通路促进人卵巢癌细胞发生EMT。     

Chrysin inhibits metastatic potential of human triple‐negative breast cancer cells by modulating matrix metalloproteinase‐10, epithelial to mesenchymal transition,and PI3K/Akt signaling pathway

Chrysin, a naturally occurring flavone, has been shown to inhibit cell proliferation and induce cell apoptosis in various cancers. However, the effect and mechanisms of chrysin on cancer metastasis are still enigmatic. In this study, metastatic triple‐negative breast cancer (TNBC) cell lines were used to evaluate the antimetastatic activity of chrysin. The results showed that chrysin (5, 10 and 20 μM) significantly suppressed TNBC cell migration and invasion in a dose‐dependent manner. Human matrix metalloproteinase (MMP) antibody array demonstrated that MMP‐10 was downregulated by chrysin, which was further verified by Western blotting and ELISA. Moreover, it was shown that chrysin induced increased E‐cadherin expression and decreased expression of vimentin, snail and slug in TNBC cells, suggesting that chrysin had a reversal effect on epithelial–mesenchymal transition. More importantly, it was demonstrated that inhibiting the Akt signal pathway might play a central role in chrysin‐induced antimetastatic activity by regulating MMP‐10 and epithelial–mesenchymal transition. In conclusion, our study indicates that chrysin exerts antimetastatic activities in TNBC cells, which suggests that chrysin might be a potential therapeutic candidate for the treatment of advanced or metastatic breast cancer. Copyright © 2013 John Wiley & Sons, Ltd.     

白藜芦醇调控NLRP3炎症小体介导的血管平滑肌细胞焦亡抗动脉粥样硬化研究

目的 探讨白藜芦醇对动脉粥样硬化（AS）中血管钙化的影响及机制。方法 将雄性健康SD大鼠随机分为对照组、模型组、白藜芦醇（5 mg·kg^-1）组,每组10只。造模前ip预给药21 d,每天1次。模型组、白藜芦醇组均制备AS模型：sc 5 mg·kg^-1维生素D₃5 d后,分离并结扎左颈动脉,0.12 mmol·L^-1的CaCl₂溶液湿敷30 min,随即缝合,对照组湿敷生理盐水。造模后继续给药,1周后所有大鼠同时禁食24 h后处死,取大鼠左颈动脉进行苏木精-伊红（HE）染色和Von Kossa染色。体外钙化培养基诱导CRL-1999细胞钙化,给予白藜芦醇（10μmol·L^-1）干预,茜素红S染色检测钙盐沉积;实时荧光定量PCR （qRT-PCR）和Western blotting法检测钙化指标骨形态发生蛋白-2（BMP2）、侏儒相关转录因子2（Runx2）,炎性小体NOD样受体热蛋白结构域相关蛋白3（NLRP3）,细胞焦亡相关指标天冬氨酸蛋白水解酶-1（Caspase-1）、Gasdermin-D （GSDMD）、白细胞介素（IL）-1β、IL-18蛋白和mRNA表达变化;免疫荧光检测Runx2、NLRP3、Caspase-1、GSDMD、IL-1β蛋白表达;分子对接验证白藜芦醇与靶蛋白NLRP3、Caspase-1、GSDMD、BMP2、Runx2结合活性。结果 HE与VonKossa染色显示模型组血管壁结构紊乱、钙盐沉积,白藜芦醇缓解钙盐沉积。体外实验表明钙化结节可被白藜芦醇缓解;与对照组比较,模型组BMP2、Runx2、NLRP3、Caspase-1、GSDMD、IL-1β、IL-18 mRNA和蛋白表达均显著上升（P<0.05、0.01、0.001）;经白藜芦醇处理后,上述指标的mRNA和蛋白表达均显著下降（P<0.05、0.01、0.001）。白藜芦醇与Runx2、NLRP3分别形成2、3个氢键,对接活性较好。结论 白藜芦醇抑制AS中血管钙化,机制可能与抑制NLRP3/Caspase-1信号通路,进而抑制细胞焦亡有关。     

Cross-regulation between protein L-isoaspartyl O-methyltransferase and ERK in epithelial mesenchymal transition of MDA-MB-231 cells

Aim:

Protein L-isoaspartyl O-methyltransferase (PIMT) regulates cell adhesion in various cancer cell lines through activation of integrin αv and the PI3K pathway. The epithelial mesenchymal transition (EMT) enables epithelial cells to acquire the characteristics of mesenchymal cells, and to allow them to migrate for metastasis. Here, we examined the relationship between PIMT and EMT with attached or detached MDA-MB 231 cells.

Methods:

Human breast cancer cell line MDA-MB-231 cells were maintained in a suspension on poly-HEMA in the presence or absence of PIMT siRNA or ERK inhibitor PD98059. The mRNAs and proteins were analyzed using RT-PCR and immunoblotting, respectively.

Results:

During cellular incubation under detached conditions, PIMT, integrin αv and EMT proteins, such as Snail, Slug and matrix metalloproteinase 2 (MMP-2), were significantly increased in correlation with the phosphorylation of ERK1/2. The ERK inhibitor PD98059 (25 μmol/L) strongly suppressed the expression of the proteins and PIMT. Interestingly, PIMT siRNA blocked the phosphorylation of ERK and the expression of the EMT proteins. Additionally, PIMT and ERK phosphorylation were both co-activated by treatment with TGF-β (10 ng/mL) and TNF-α (10 ng/mL).

Conclusion:

A tight cross-regulation exists between ERK and PIMT in regards to their activation and expression during the EMT.     

Wnt信号通路诱导肿瘤细胞上皮间质转化的研究进展

肿瘤细胞发生上皮间质转化(epithelial-mesenchymaltransition,EMT)从而具有侵袭转移的能力,是肿瘤发展的一个重要过程。既往研究发现Wnt信号通路调节控制着许多生命过程,也发现其是诱导EMT不可缺少的重要通路。通过对Wnt信号分子的调控,可以有效的阻断甚至逆转EMT。Wnt诱导EMT发生发展过程的研究,也为抗肿瘤新药开发提供了新思路,推动了药物的研发。该文旨在对近年Wnt信号通路诱导EMT的研究做一综述。     

青藤碱通过调控PI3K/AKT通路介导的上皮间质转化抑制胰腺癌AsPC-1细胞侵袭和转移

目的 探讨青藤碱对胰腺癌AsPC-1细胞上皮间质转化（epithelial mesenchymal transition,EMT）、侵袭和转移的影响及机制。方法 采用不同浓度青藤碱作用胰腺癌AsPC-1细胞后,划痕愈合试验检测细胞愈合能力,Transwell试验检测迁移细胞数和侵袭细胞数,Western blotting检测E-cadherin、N-cadherin、Vimentin、PI3K、p-PI3K、AKT、p-AKT蛋白表达。结果 青藤碱降低AsPC-1细胞相对迁移率、迁移细胞数、侵袭细胞数。青藤碱上调E-cadherin蛋白表达、下调N-cadherin、Vimentin蛋白表达。青藤碱降低p-PI3K/PI3K、p-AKT/AKT比值。胰岛素生长因子-1逆转青藤碱对E-cadherin蛋白表达的上调和对N-cadherin、Vimentin蛋白表达的下调作用。TGF-β逆转青藤碱对AsPC-1细胞相对迁移率、迁移细胞数、侵袭细胞数的降低作用。结论 青藤碱通过调控PI3K/AKT通路介导的EMT,抑制AsPC-1细胞侵袭和转移。     

NLRP3 inflammasome activation involved in LPS and coal tar pitch extract‐induced malignant transformation of human bronchial epithelial cells

Inflammatory microenvironment has been found as a new characteristic of cancer; however, the mechanisms of inflammation‐related lung cancer remain unclear. To explore the role of NLRP3 inflammsome activation in inflammation‐related lung carcinogenesis, a cell model was set up. Human bronchial epithelial cells (BEAS‐2B) were stimulated with 1 μg/mL lipopolysaccharide (LPS) for 24 hours, and then treated with 2.4 μg/mL coal tar pitch extract (CTPE) for 24 hours, after removal of LPS and CTPE, the cells were numbered passage 1 and were passaged and treated in this way until passage 30, which was called LPS + CTPE group. DMSO and Saline were used as vehicle controls. Malignant transformation of cells in passage 30 was evaluated by morphological change, platelet clone formation assay, and tumor formation in nude mice. The mRNA levels of NLRP3 and IL‐1β were detected by real time‐PCR. The combination of NLRP3 and caspase‐1 were determined using immunofluorescence and confocal. The protein expression of NLRP3, cleaved caspase‐1(p10), and cleaved IL‐1β was detected using Western blot. It was shown that CTPE, LPS + CTPE‐stimulated BEAS‐2B cells of passage 30 changed a lot morphologically. The clone formation rates, the rates of positive cells of NLRP3 and caspase‐1 combination, the mRNA levels of NLRP3 and IL‐1β, the protein expression of NLRP3, cleaved caspase‐1(p10) and cleaved IL‐1β of cells exposed with CTPE and LPS + CTPE at passage 30 were significantly increased compared to vehicle controls. Furthermore, the ability of tumor formation in nude mice, the rates of clone formation and positive cells, mRNA and protein levels of NLRP3 inflammasome activation‐related factors in LPS + CTPE‐induced cells were all higher than those in cells stimulated with CTPE alone. In conclusion, the cell model of inflammation‐related lung cancer is set up successfully, and NLRP3 inflammasome activation may be involved in the malignant transformation of BEAS‐2B cells which induced by CTPE alone or LPS combined with CTPE.     

Activation of the inflammasome by amorphous silica and TiO2 nanoparticles in murine dendritic cells

Abstract

Nanomaterials are increasingly used in various food applications. In particular, nanoparticulate amorphous SiO₂ is already contained, e.g., in spices. Since intestinal dendritic cells (DC) could be critical targets for ingested particles, we compared the in vitro effects of amorphous silica nanoparticles with fine crystalline silica, and micron-sized with nano-sized TiO₂ particles on DC. TiO₂- and SiO₂-nanoparticles, as well as crystalline silica led to an upregulation of MHC-II, CD80, and CD86 on DC. Furthermore, these particles activated the inflammasome, leading to significant IL-1β-secretion in wild-type (WT) but not Caspase-1- or NLRP3-deficient mice. Silica nanoparticles and crystalline silica induced apoptosis, while TiO₂ nanoparticles led to enhanced production of reactive oxygen species (ROS). Since amorphous silica and TiO₂ nanoparticles had strong effects on the activation-status of DC, we suggest that nanoparticles, used as food additives, should be intensively studied in vitro and in vivo, to ensure their safety for the consumer.     

Nerve growth factor induces cord formation of mesenchymal stem cell by promoting proliferation and activating the PI3K/Akt signaling pathway

Aim:

To investigate whether nerve growth factor (NGF) induced angiogenesis of bone marrow mesenchymal stem cells (MSCs) and the underlying mechanisms.

Methods:

Bone marrow MSCs were isolated from femors or tibias of Sprague-Dawley rat, and cultured. The cells were purified after 3 to 5 passages, seeded on Matrigel-coated 24-well plates and treated with NGF. Tube formation was observed 24 h later. Tropomyosin-related kinase A (TrkA) and p75NTR gene expression was examined using PCR analysis and flow cytometry. Growth curves were determined via cell counting. Expression of VEGF and pAkt/Akt were analyzed with Western blot.

Results:

NGF (25, 50, 100 and 200 μg/L) promoted tube formation of MSCs. The tubular length reached the maximum of a 2.24-fold increase, when the cells were treated with NGF (50 μg/L). NGF (50 μg/L) significantly enhanced Akt phosphorylation. Pretreatment with the specific PI3K inhibitor {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002 (10 μmol/L) blocked NGF-stimulated Akt phosphorylation, tube formation and angiogenesis. NGF (25–200 μg/L) did not affect the expression of TrkA and vascular endothelial growth factor (VEGF), but significantly suppressed the expression of p75NTR. NGF (50 μg/L) markedly increased the proliferation of MSCs.

Conclusion:

NGF promoted proliferation of MSCs and activated the PI3K/Akt signaling pathway, which may be responsible for NGF induction of MSC angiogenesis.     

猪苓多糖通过JAK2-STAT3通路抑制N-甲基-N''-硝基-N-亚硝基诱导的GES-1细胞上皮间充质转化

目的 研究猪苓多糖抑制胃黏膜上皮细胞（GES-1）间充质转化的作用和机制。方法 应用N-甲基-N''-硝基-N-亚硝基（MNNG,40 μmol·L^-1）诱导GES-1细胞间充质转化模型,造模同时给予猪苓多糖（1.25、2.50、5.00、10.00 mg·mL^-1）处理24、48 h后,CCK-8法检测细胞增殖能力;造模同时给予猪苓多糖（5.00 mg·mL^-1）处理48 h后,实时定量PCR（qRT-PCR）法检测猪苓多糖对N-cadherin、Snail、Twist、Fibronection及Vmention mRNA表达的影响;Western blotting法检测E-cadherin、N-cadherin、Snail、Twist、Fibronetin、Vimentin、STAT3、p-STAT3、JAK2、p-JAK2蛋白表达。裸鼠分别sc经MNNG 40 μmol·L^-1处理48 h后的GES-1细胞（5×10⁷·mL^-1 ,0.2 mL）、胃癌细胞SGC7901（5×10⁷·mL^-1 ,0.2 mL ,阳性对照）,1个月后观察各组成瘤及体质量情况,注射局部进行HE染色后行组织病理学观察,验证GES-1细胞经MNNG处理后是否达到肿瘤阶段。结果 与模型组比较,随猪苓多糖浓度的增加,GES-1细胞增殖能力逐渐上升,到达5 mg·mL^-1时增殖能力最高,差异显著（P<0.01、0.001）;猪苓多糖组N-cadherin、Snail、Twist、Fironetion及Vimentin的mRNA表达均显著下降（P<0.05、0.01、0.001）;猪苓多糖组E-cadherin蛋白表达显著上升（P<0.05）,N-cadherin、Snail、Twist、Fibronetion、Vimentin及通路蛋白STAT3、p-STAT3、JAK2、p-JAK2表达显著下降（P<0.05、0.01、0.001）。对照组和MNNG处理的GES-1组未见瘤体形成,阳性对照SGC7901组注射后1周左右局部出现瘤体,1个月瘤体长至6 cm左右;MNNG处理的GES-1组裸鼠精神较差,体质量明显减轻,SGC7901组裸鼠状态差,体质量大幅减轻;对照组和MNNG组病理染色显示为正常的皮肤组织,阳性对照组显示为肿瘤组织。结论 猪苓多糖可能通过抑制JAK2-STAT3通路干预MNNG诱导的GES-1细胞上皮间充质化。     

The mechanisms for lung cancer risk of PM2.5: Induction of epithelial‐mesenchymal transition and cancer stem cell properties in human non‐small cell lung cancer cells

Fine particulate matter (PM_2.5) is a major component of air pollutions that are closely associated with increased risk of lung cancer. However, the role of PM_2.5 in the etiology of lung cancer is largely unknown. In this study, we performed acute (24 hours) and chronic (five passages) exposure models to investigate the carcinogenetic mechanisms of PM_2.5 by targeting the induction of epithelial‐mesenchymal transition (EMT) and cancer stem cells (CSC) properties in human non‐small cell lung cancer cell line A549. We found that both acute and chronic PM_2.5 exposure enhanced cell migration and invasion, decreased mRNA expression of epithelial markers and increased mRNA expression of mesenchymal markers. Chronic PM_2.5 exposure further induced notable EMT morphology and CSC properties, indicating the developing process of cell malignant behaviors from acute to chronic PM_2.5 exposure. CSC properties induced by chronic PM_2.5 exposure characterized with increased cell‐surface markers (CD44, ABCG2), self‐renewal genes (SOX2 and OCT4), side population cells and neoplastic capacity. Furthermore, the levels of three stemness‐associated microRNAs, Let‐7a, miR‐16 and miR‐34a, were found to be significantly downregulated by chronic PM_2.5 exposure, with microarray data analysis from TCGA database showing their lower expression in human lung adenocarcinoma tissues than that in the adjacent normal lung tissues. These data revealed that the induction of EMT and CSC properties were involved in the lung cancer risk of PM_2.5, and implicated CSC properties and related microRNAs as possible biomarkers for carcinogenicity prediction of PM_2.5.     

紫草素对肺炎链球菌引起的肺炎小鼠细胞外信号调节激酶 / p38丝裂素活化蛋白激酶 /核苷酸结合寡聚化结构域样蛋白 3信号通路及肺血管通透性的影响

目的 探究紫草素对肺炎链球菌引起的肺炎小鼠细胞外信号调节激酶/p38丝裂素活化蛋白激酶/核苷酸结合寡聚化结构域样蛋白3(ERK/p38/NLRP3)信号通路及肺血管通透性的影响.方法 采用肺炎链球菌悬液50 L滴加BALB/c小鼠破损鼻黏膜建立肺炎链球菌性肺炎模型,成模小鼠采用随机数字表法分为模型组、紫草素低、中、高浓度组及头孢呋辛酯组,另取鼻腔滴加50 L生理盐水BALB/c小鼠为对照组,每组10只,紫草素低、中、高浓度组分别给予紫草素12.5 mg/kg、25.0 mg/kg、50.0 mg/kg,头孢呋辛酯组给予头孢呋辛酯50.0 mg/kg,对照组、模型组给予等量生理盐水,每天1次,连续灌胃7 d.末次给药12 h后处死小鼠,检测各组小鼠左肺湿/干重比值(W/D),每组采用随机数字表法选取4只小鼠进行1％伊文氏蓝5 mL/kg尾静脉注射检测肺血管通透性,酶联免疫吸附试验(ELISA)检测肺组织白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)表达量,实时荧光定量聚合酶链式反应(RT-qPCR)及蛋白质印迹法(WB)分别检测肺组织ERK、p38、NLRP3 mRNA及蛋白表达.结果 与对照组比较,模型组小鼠肺组织W/D比值[(4.49±0.27)％比(3.02±0.15)％]、肺血管通透性[(0.091±0.009)g/mg比(0.034±0.004)g/mg]、IL-1β[(567.29±14.65)pg/mL比(102.46±6.73)pg/mL]、TNF-α[(417.16±12.89)pg/mL比(83.59±6.28)pg/mL]、ERK、p38、NLRP3 mRNA及蛋白表达水平均显著增加(P<0.05);与模型组比较,紫草素低、中、高浓度组小鼠肺组织W/D比值、肺血管通透性、IL-1β、TNF-α表达量、ERK、p38、NLRP3 mRNA及蛋白表达水平均依次降低(P<0.05),紫草素高剂量组均低于头孢呋辛酯组(P<0.05).结论 紫草素可减轻肺炎链球菌性肺炎小鼠肺组织损伤、炎症反应及肺血管通透性,可能与抑制ERK/p38/NL?RP3信号通路有关.     

溪黄草水提物对高糖诱导的大鼠HBZY-1细胞炎症反应和氧化应激的抑制作用以及对TLR 4/NF-κB/NLRP 3通路的作用

目的：溪黄草水提物（RWE）对高糖诱导的大鼠HBZY-1细胞炎症反应和氧化应激的抑制作用以及对TLR 4/NF-κB/NLRP 3通路的作用。方法：用高糖（HG,30 mmol·L^-1）孵育大鼠肾小球细胞株HBZY-1细胞,MTT法测定不同浓度RWE（5,10,20 mg·mL^-1）对HG环境下HBZY-1细胞增殖的影响,测定RWE对HG诱导的HBZY-1细胞氧化应激水平和炎性细胞因子表达的影响,同时测定其对细胞TLR4/NF-κB/NLRP3信号通路的影响。结果：HG能诱导HBZY-1细胞增殖,提高MDA、IL-6、IL-1β、TNF-α水平和TLR 4/NF-κB/NLRP 3信号通路的表达,降低SOD和GSH水平;RWE减弱细胞的增殖能力,降低MDA、IL-6、IL-1β和TNF-α的水平,提高SOD和GSH水平,抑制TLR 4/NF-κB/NLRP 3信号通路的表达。结论：RWE对HG诱导的HBZY-1细胞具有保护作用,其作用机制主要是抑制炎症反应和氧化应激,TLR 4/NF-κB/NLRP 3信号通路可能参与RWE对细胞的保护作用。     

Mangiferin inhibits lipopolysaccharide‐induced epithelial‐mesenchymal transition (EMT) and enhances the expression of tumor suppressor gene PER1 in non‐small cell lung cancer cells

Non‐small cell lung cancer (NSCLC) is often complicated by pulmonary infection, which affects treatment and prognosis. Bacterial lipopolysaccharide (LPS) is an effective stimulator of inflammatory cytokine production, and previous studies have reported that LPS promotes tumor invasion and metastasis. Mangiferin is a plant‐derived C‐glucosylxanthone with many biological activities, such as antioxidation and anti‐inflammation. This research mainly explored the mechanism of its antitumor activities on LPS‐induced A549, NCI‐H460, and NCI‐H520 NSCLC cells. We determined that mangiferin exhibits growth inhibiting activity against LPS‐induced NSCLC cells through the 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide (MTT) assay. In addition, mangiferin reversed the LPS‐induced downregulation of E‐cadherin (epithelial marker); conversely, it significantly inhibited the expression of raised vimentin (mesenchymal markers). Moreover, the ability of NSCLC cells to migrate, as evidenced by the wound healing and transwell migration assays, and the expression of CXCR4 increased by LPS were significantly repressed by mangiferin. In addition, mangiferin markedly mediated protein levels of PER1 and NLRP3 in LPS‐induced NSCLC cells and reduced the secretion of IL‐1β. These results indicate that mangiferin is not only a remarkable anti‐inflammatory compound but also an antitumor agent; thus, it has the potential for being developed into anti‐inflammatory and antitumor drugs in the future.