Neferine inhibits the progression of diabetic nephropathy by modulating the miR-17-5p/nuclear factor E2-related factor 2 axis

OBJECTIVE:To investigate the effect of Neferine(Nef)on diabetic nephropathy(DN)and to explore the mechanism of Nef in DN based on miRNA regulation theory.METHODS:A DN mouse model was constructed and treated with Nef.Serum creatinine(Crea),blood urea(UREA)and urinary albumin were measured in mice by kits,and renal histopathological changes and fibrosis were observed by hematoxylin-eosin staining and Masson staining.Renal tissue superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione peroxidase(GSH-Px)activities were measured by enzyme-linked immunosorbent assay(ELISA).Western blotting was used to detect the expression of nuclear factor E2-related factor 2(Nrf2)/heme oxygenase 1(HO-1)signaling pathway-related proteins in kidney tissues.Quantitative reverse transcription-polymerase chain reaction(q RT-PCR)was used to detect the expression of miR-17-5p in kidney tissues.Subsequently,a DN in vitro model was constructed by high glucose culture of human mesangial cells(HMCs),cells were transfected with miR-17-5p mimic and/or treated with Nef,and we used q RTPCR to detect cellular miR-17 expression,flow cytometry to detect apoptosis,ELISAs to detect cellular SOD,MDA,and GSH-Px activities,Western blots to detect Nrf2/HO-1 signaling pathway-related protein expression,and dual luciferase reporter gene assays to verify the targeting relationship between Nrf2 and miR-17-5p.RESULTS:Administration of Nef significantly reduced the levels of blood glucose,Crea,and UREA and the expression of miR-17-5p,improved renal histopathology and fibrosis,significantly reduced MDA levels,elevated SOD and GSH-Px activities,and activated Nrf2 expression in kidney tissues from mice with DN.Nrf2 is a post-transcriptional target of miR-17-5p.In HMCs transfected with miR-17-5p mimics,the m RNA and protein levels of Nrf2 were significantly suppressed.Furthermore,miR-17-5p overexpression and Nef intervention resulted in a significant increase in high glucose-induced apoptosis and MDA levels in HMCs and a significant decrease in the protein expression of HO-1 and Nrf2.CONCLUSION:Collectively,these results indicate that Nef has an ameliorative effect on DN,and the mechanism may be through the miR-17-5p/Nrf2 pathway.

Microwave absorption properties regulation and bandwidth formula of oriented Y_(2)Fe_(17)N_(3-δ)@SiO_(2)/PU composite synthesized by reduction-diffusion method

As concepts closely related to microwave absorption properties,impedance matching and phase matching were rarely combined with material parameters to regulate properties and explore related mechanisms.In this work,reduction–diffusion method was innovatively applied to synthesize rare earth alloy Y_(2)Fe_(17).In order to regulate the electromagnetic parameters of absorbers,the Y_(2)Fe_(17)N_(3-δ)particles were coated with silica(Y_(2)Fe_(17)N_(3-δ)@SiO_(2))and absorbers with different volume fractions were prepared.The relationship between impedance matching,matching thickness,and the strongest reflection loss peak(RLmin)was presented obviously.Compared to the microwave absorption properties of Y_(2)Fe_(17)N_(3-δ)/PU absorber,Y_(2)Fe_(17)N_(3-δ)@SiO_(2)/PU absorbers are more conducive to the realization of microwave absorption material standards which are thin thickness,light weight,strong absorbing intensity,and broad bandwidth.Based on microwave frequency bands,the microwave absorption properties of the absorbers were analyzed and the related parameters were listed.As an important parameter related to perfect matching,reflection factor(√ε_(r)/μ_(r))was discussed combined with microwave amplitude attenuation.According to the origin and mathematical model of bandwidth,the formula of EAB(RL<-10 dB)was derived and simplified.The calculated bandwidths agreed well with experimental results.

KLF17对裸鼠移植瘤生长的影响及其在体调控靶基因的筛选和功能分析

目的:研究Krüppel样因子17(Krüppel-like factor 17,KLF17)在裸鼠移植瘤中的作用并分析KLF17在体调控的靶基因及其功能和参与的信号通路。方法:慢病毒转染技术稳定上调人肺腺癌A549细胞及下调人肺腺癌H322细胞中KLF17的表达。11只BLAB/c nu/nu裸鼠分为上调组5只和下调组6只,分别通过左、右侧躯体皮下注射KLF17上调和下调的相应细胞及其对照细胞,观察KLF17对裸鼠移植瘤生长的影响;Real-time PCR及免疫组织化学染色分析裸鼠移植瘤组织中KLF17 mRNA及蛋白的表达,转录组测序技术分析上调KLF17表达后裸鼠移植瘤中差异表达的基因,通过The Cancer Genome Atlas(TCGA)数据库分析KLF17调控的差异基因的功能,Gene Ontology和KEGG PATHWAY富集分析差异基因的功能和可能参与的信号通路。结果 :上调A549细胞中KLF17的表达后,其裸鼠移植瘤生长速率显著低于空载体对照组(P<0.05),下调H322细胞中KLF17表达后,其裸鼠移植瘤生长速率及移植瘤重量均显著高于空载体对照组(P<0.01,P<0.05)。在裸鼠移植瘤组织中,过表达组KLF17 mRNA及蛋白显著高于对照空载体组。转录组测序结果显示KLF17可能调控的基因有ras基因同源家族成员V(ras homolog family member V,RHOV)和冠蛋白1C(coronin 1C,CORO1C)等。TCGA数据库中肺腺癌患者10年累计生存时间在RHOV和CORO1C mRNA不同表达组明显不同,高表达RHOV及CORO1C的肺腺癌患者生存时间显著低于其低表达组。Gene Ontology和KEGG PATHWAY富集分析提示KLF17调控的靶基因(差异基因)可能与肿瘤细胞的刺激应答、生长及黏附有关,并且参与了细胞趋化性、黏附及细胞外基质受体相关的信号通路。结论:KLF17抑制裸鼠移植瘤的生长,并下调RHOV和CORO1C等癌基因,参与调控肿瘤黏附及生长相关信号通路。

Hypoxia activates the hypoxia-inducible factor-1α/vascular endothelial growth factor pathway in a prostatic stromal cell line:A mechanism for the pathogenesis of benign prostatic hyperplasia

Background The development of benign prostatic hyperplasia(BPH)is closely related to hypoxia in the prostatic stroma,and the hypoxia-inducible factor-1α/vascular endothelial growth factor(HIF-1α/VEGF)pathway has been shown to significantly activate in response to hypoxia.The underlying mechanism for activation of this pathway in the pathogenesis of BPH remains unclear.Materials and methods We constructed HIF-1αoverexpression and knockdown BPH stromal(WPMY-1)and epithelial(BPH-1)cell lines,which were cultured under different oxygen conditions(hypoxia,normoxia,and hypoxia+HIF-1αinhibitor).Quantitative real-time polymerase chain reaction(qPCR)and Western blotting were applied to detect the expression of the HIF-1α/VEGF pathway.Cell proliferation and apoptosis were analyzed by Cell Counting Kit-8 and flow cytometry.We used the miRWalk 2.0 database and Western blotting to predict the potential miRNA that selectively targets the HIF-1α/VEGF pathway,and verified the prediction by qPCR and dual-luciferase assays.Results In a BPH stromal cell line(WPMY-1),the expression of VEGF was in accordance with HIF-1αlevels,elevated in the overexpression cells and decreased in the knockdown cells.Hypoxia-induced HIF-1αoverexpression,which could be reversed by a HIF-1αinhibitor.Moreover,the HIF-1αinhibitor significantly depressed cellular proliferation and promoted apoptosis in hypoxic conditions,assessed by Cell Counting Kit-8 and flow cytometry.However,in the BPH epithelial cell line(BPH-1),the expression level of HIF-1αdid not influence the expression of VEGF.Finally,a potential miRNA,miR-17-5p,regulating the HIF-1α/VEGF pathway was predicted from the miRWalk 2.0 database and Western blotting,and verified by qPCR and dual-luciferase assay.Conclusions In hypoxia,activation of the HIF-1α/VEGF pathway plays a crucial role in regulating cell proliferation in a BPH stromal cell line.Regulation by miR-17-5p may be the potential mechanism for the activation of this pathway.Regulation of this pathway may be involved in the pathogenesis of BPH.

Kruppel样因子17抑制肾癌786-0细胞迁移和侵袭及机制

目的探讨Kruppe1样因子17（KLF17）是否通过调控上皮-间充质转化（EMT）抑制786-0细胞的迁移侵袭能力。方法设计并合成特异性小干扰RNA（siRNA）下调肾癌细胞株786-0中KLF17的表达，并利用划痕实验及Transwell实验检测其迁移侵袭能力的变化，通过Westernblot检测EMT相关分子表达水平变化。结果siRNA能有效下调肾癌786-0细胞株中KLFl7的蛋白及mRNA的表达量。细胞划痕实验结果显示下调KLF17表达后，细胞迁移能力增强（P〈0．01）；Transwell实验中阴性对照组穿膜细胞数为（18．16±2．21）个，而siRNA1组为（29．03±2．48）个（P〈0．01），siRNA2组为（28．93±3．00）个（P〈0．01）。KLF17-siRNA介导的基因沉默下调了上皮细胞表面标记E-钙黏蛋白（E-cadberin）、β-连环蛋白（β-catenin）的表达，上调了间充质干细胞表面标记N-钙黏蛋白（N-cadherin）的表达。结论KLF17可通过调节EMT的进程影响肾癌细胞的迁移侵袭能力。