糖尿病视网膜病变中视网膜内皮细胞功能障碍的研究进展北大核心

视网膜内皮细胞(REC)是参与糖尿病视网膜病变和许多眼部疾病的主要细胞类型之一。视网膜微血管系统有助于血-视网膜屏障的维持,这对正常的视功能至关重要。REC的改变在视网膜疾病的发生发展中起着关键作用。高血糖是糖尿病微血管损伤的重要原因,通过不同的机制导致REC功能障碍,包括向衰老表型改变、迁移和增殖能力增强、炎性凋亡等,最终导致无细胞毛细血管及病理性新生血管形成。本文对糖尿病视网膜病变中REC的功能障碍作一综述。     

脑出血继发损伤中沉默信息调节因子2的表达和炎症反应*

目的：探讨脑出血对酵母沉默信息调节因子2（Sirt2）和炎症的影响。方法：将胶原酶Ⅳ注入SD大鼠右侧 纹状体中建立脑出血模型，通过免疫印迹和ELISA 等方法测定大鼠脑出血后48 h 的Sirt2 的表达及炎症变化。利 用Hemin 诱导PC12 细胞损伤模拟体外脑出血模型，并检测Sirt2 及炎症变化；采用短发夹RNA（shRNA）-Sirt2 沉 默Sirt2 在PC12 细胞中的表达及对炎症的影响。结果：手术后48 h 脑出血行为学评分最低。脑出血组Sirt2 的表达 显著高于假手术组。脑出血组IL-6、IL-1β 表达显著升高。结论：脑出血可以促进Sirt2 的表达和炎症反应，降低 Sirt2 的表达可减缓炎症反应。 关键词 脑出血；沉默信息调节     

基于网络药理学探讨搜风愈喘方拆方“祛宿痰方”调控儿童哮喘的作用机制＊

目的 基于“伏风暗瘀宿痰”小儿哮喘病机新说，采用网络药理学及实验验证的方法，探索搜风愈喘方拆方“祛宿痰方”治疗儿童哮喘的作用机制，验证中医“宿痰”病机与西医细胞外基质改变病理之间的交通性。方法 通过TCMSP数据库建立“祛宿痰方”的有效成分和靶点数据集，利用OMIM、GeneCards、DrugBank、TTD疾病数据库建立哮喘疾病靶点数据集，利用Cytoscape软件取交集并构建“祛宿痰方”与哮喘的蛋白质互作网络，筛选关键靶点蛋白。利用Metascape数据库进行基因本体（Gene ontology，GO）分析以及京都基因和基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析。复制卵蛋白（OVA）诱导的哮喘大鼠模型，对核心通路及关键靶点进行实验验证。结果 共得到“祛宿痰方”治疗哮喘的靶点98个，包括IL-13、TP53、TGF-β1、VEGF-A、MMP9等，KEGG得到与哮喘相关的通路287条（P＜0.05），包括NF-κB信号通路、PI3K-AKT信号通路、IL-17信号通路等。GO结果显示与哮喘相关生物进程包括炎症反应、细胞外基质调控、氧化应激、血管生成等。动物实验证实“祛宿痰方”可下调大鼠肺组织中p-NF-κB-P65磷酸化水平，抑制NF-κB信号通路的激活，降低IL-13、TGF-β1 mRNA表达量（P<0.05），减少哮喘大鼠肺组织中炎症细胞浸润、黏液产生，从而延缓哮喘的进程。结论 “祛宿痰方”可抑制NF-κB信号通路的激活，降低肺组织中IL-13、TGF-β1 mRNA表达量，可能通过抑制炎症反应、调控细胞外基质等途径作用于哮喘，中医“宿痰”病机与西医细胞外基质改变病理之间存在一定的的交通性。     

Inhibition of autophagy enhances apoptosis induced by bortezomib in AML cells

Bortezomib is a novel proteasome inhibitor, which has been successfully used to treat mantle cell lymphoma and multiple myeloma. However, the direct effects of bortezomib on acute promyelocytic leukaemia (APL) have not been fully investigated. In the present study, the WST-8 assay, western blotting, flow cytometry, monodansylcadaverine staining and transmission electron microscopy were performed. It was demonstrated that bortezomib treatment induced a time- and dose-dependent decrease in the viability of NB4 cells. Bortezomib treatment induced cell apoptosis in NB4 cells, as assessed by Annexin V/propidium iodide analysis, and the detection of cleaved caspase-3, cleaved poly(ADP-ribose) polymerase, Bax and Bcl-2 expression. Furthermore, bortezomib treatment induced autophagy in NB4 cells, as indicated by autophagosome formation, p62 degradation, LC3-I to LC3-II conversion and formation of acidic autophagic vacuoles. Notably, autophagy induced by bortezomib was initiated prior to apoptosis. Inhibition of autophagy by knocking down Beclin-1 expression increased bortezomib-induced apoptosis in NB4 cells. Therefore, the present study revealed that the combination of bortezomib and autophagy inhibition may be a potential treatment strategy for APL.     

Silencing KLF16 inhibits oral squamous cell carcinoma cell proliferation by arresting the cell cycle and inducing apoptosis

Krüppel-like factor 16 (KLF16), a member of the Krüppel-like factor (KLF) family, has been extensively investigated in multiple cancer types. However, the role of KLF16 in oral squamous cell carcinoma (OSCC) remains unknown. Thus, we conducted this study to investigate its related mechanism. KLF16 expression in OSCC cell lines was quantified by western blotting. Then, OECM1 and OC3 cells were divided into Blank, siCtrl, siKLF16#1 and siKLF16#2 groups. Subsequently, cell proliferation was detected using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assays, cell migration and invasion were detected with wound healing and Transwell assays, and cell cycle distribution and cell apoptosis were detected via flow cytometry. KLF16, p21, CDK4, Cyclin D1 and p-Rb expression was detected by western blotting. Finally, xenograft models were established in nude mice to observe the in vivo effects of KLF16 on OSCC. KLF16 protein expression was upregulated in OSCC cells. Compared to the cells in the Blank group, the OECM1 and OC3 cells in the siKLF16#1 group and siKLF16#2 group exhibited a sharp decrease in proliferation but a remarkable increase in apoptosis. Moreover, the proportion of cells in the G0/G1 phase notably increased and that in the S phase decreased, with evident decreases in cell invasion and migration. Moreover, KLF16, cyclin-dependent kinase 4 (CDK4), Cyclin D1 and p-Rb protein expression was upregulated, but p21 expression was downregulated. The mice in the siKLF16#1 and siKLF16#2 xenograft model groups exhibited slower tumour growth and smaller tumours with evident downregulation of Ki67 expression compared to the mice in the Blank group. KLF16 expression was upregulated in OSCC cells, and interfering with KLF16 led to cell cycle arrest, inhibited OSCC cell growth and promoted cell apoptosis.     

A basal‐enriched microRNA is required for prostate tumorigenesis in a Pten knockout mouse model

MicroRNAs (miRNAs) play important roles in prostate cancer development. However, it remains unclear how individual miRNAs contribute to the initiation and progression of prostate cancer. Here we show that a basal layer‐enriched miRNA is required for prostate tumorigenesis. We identify miR‐205 as the most highly expressed miRNA and enriched in the basal cells of the prostate. Although miR‐205 is not required for normal prostate development and homeostasis, genetic deletion of miR‐205 in a Pten null tumor model significantly compromises tumor progression and does not promote metastasis. In Pten null basal cells, loss of miR‐205 attenuates pAkt levels and promotes cellular senescence. Furthermore, although overexpression of miR‐205 in prostate cancer cells with luminal phenotypes inhibits cell growth in both human and mouse, miR‐205 has a minimal effect on the growth of a normal human prostate cell line. Taken together, we have provided genetic evidence for a requirement of miR‐205 in the progression of Pten null‐induced prostate cancer.     

Ginsenoside Rg1 prevents acetaminophen-induced oxidative stress and apoptosis via Nrf2/ARE signaling pathway

Inappropriate use of acetaminophen (APAP) can lead to morbidity and mortality secondary to hepatic necrosis. Ginsenoside Rg1 is a major active ingredient in processed Panax ginseng, which is proved to elicit biological effects. We hypothesized the beneficial effect of Rg1 on APAP-mediated hepatotoxicity was through Nrf2/ARE pathway. The study was conducted in cells and mice, comparing the actions of Rg1. Rg1 significantly improved cell survival rates and promoted the expression of antioxidant proteins. Meanwhile, Rg1 reduced the excessive ROS and the occurrence of cell apoptosis, which were related to Nrf2/ARE pathway. Expression of Nrf2 has a certain cell specificity.

     

<i>Sinomenium</i> Inhibits the Viability of Hepatoma Carcinoma Cells through Activating IFNA2

Hepatocellular carcinoma (HCC) ranks the sixth place of most common cancers. Meanwhile, it is the tertiary mortality cause of cancer. There is no effective therapeutic method to prevent and treat the liver cancer. Sinomenine is a kind of Chinese traditional medicine herbal, it is reported that it can inhibit the viability of several cancer cells. The study is to explore whether sinomenine is also able to inhibit the cell viability of HCC and its potential mechanism. The IC50 of sinomenine in BEL-7402 cells was 5.351 mmol/L, and the IC50 of sinomenine in SMMC-7721 cells was 6.204 mmol/L. The gene expression results showed the relative expression of FGF2, CCND2, DCN, F3, MMP7, NRG1, HMGB1, TRIM29, HAS2, EHF, CTGF, PLK2 were down-regulated, and the relative expression of VEGF A, CITED2, NUPR1, DDX58, IRF9, NAMPT, MMP1, NDRG1, HMGA2, PPARGC1A, IFIT2, PARP9, HEY1, LOX, ETV1, ISG15, BACH, CYLD were up-regulated. Moreover, the IPA analysis results suggested that IFIT3, IFIT1, OAS1, MX1, IRF9, IFI6, IFITM1, ISG15 were up-regulated in BEL-7402 cells treated with sinomenine by activating IFNA2. The findings presented in this study may provide a promising method for the prevention and treatment of liver cancer.     

金刚丸调节Hippo信号通路mRNA表达防治OVX大鼠骨质疏松作用机制

摘要：目的 基于Hippo信号通路核心基因mRNA表达，探索具有补肾填精壮骨之效的金刚丸治疗去卵巢（ovariectomized,OVX）大鼠骨质疏松症的疗效机制。方法 通过OVX法建立绝经后骨质疏松症（postmenopausal osteoporosis,PMOP）大鼠模型，分正常组、假手术组、模型组、金刚丸高剂量组、金刚丸中剂量组、金刚丸低剂量组、仙灵骨葆对照组、骨化三醇对照组。灌胃12周后，通过X射线骨密度仪检测骨密度、镜下观察股骨头显微形态结构、ELISA法检测血清ALP、实时定量RT-PCR检测骨组织Mst2、Lats1、Taz mRNA表达。结果 ①与正常组比较，模型组股骨骨密度显著降低（P＜0.01）、骨微结构显著破坏、血清ALP显著降低（P＜0.01）、骨组织Mst2、Lats1 mRNA表达显著升高（P＜0.01）、Taz mRNA表达显著降低（P＜0.01）；②与模型组比较，除金刚丸低剂量组外，各给药组的骨密度均显著升高（P＜0.01），各给药组骨微结构破坏均得到改善、血清ALP均显著升高（P＜0.01）、骨组织Mst2、Lats1 mRNA表达均显著降低（P＜0.01）、Taz mRNA表达均显著升高（P＜0.01），均以金刚丸高剂量组最为显著。结论 骨组织Hippo信号通路核心基因Mst2、Lats1 mRNA表达上调，Taz mRNA表达下调可能是PMOP的发病机制之一；金刚丸可能通过下调骨组织Hippo信号通路核心基因Mst2、Lats1 mRNA表达、上调Taz mRNA表达的机制，有效防治PMOP。