GJB2通过AMPK/mTOR信号通路促进宫颈癌细胞恶性增殖

目的筛选宫颈癌中具有临床意义的潜在药物靶点并研究此靶点蛋白在宫颈癌发生发展中的功能和潜在分子机制。方法收集2018年7月至2019年7月济南市人民医院妇科行宫颈癌手术的30例宫颈癌患者肿瘤组织标本及癌旁组织标本,通过临床数据库分别找到在宫颈癌中高表达的基因(tumor/normal>2)以及在宫颈癌中具有显著临床预后相关性的基因(pValue<0.05),将这两部分基因进行重合分析找到具有显著临床意义的靶基因,进一步通过热图分析找到在宫颈癌中相比正常组织表达差异最明显的前4个基因,通过高表达预后差原则筛选到目的靶基因;通过siRNA介导敲低目的基因的表达,利用细胞增殖实验和划痕迁移实验验证目的基因在宫颈癌发生发展中的重要性;最后通过分析与目的基因共表达基因参与的信号通路,探索目的基因在宫颈癌中发挥功能的潜在分子机制,进一步通过蛋白免疫印迹实验进行验证。结果共有78个基因在宫颈癌中高表达且具有显著临床预后相关性;GJB2在宫颈癌中显著高表达,且高表达的患者具有不良预后;敲低GJB2抑制宫颈癌细胞的生长增殖和迁移;敲低GJB2降低AMPK的磷酸化,升高mTOR的磷酸化,可能通过AMPK/mTOR信号通路参与宫颈癌的发生发展。结论GJB2高表达可能通过抑制AMPK的磷酸化从而升高mTOR的磷酸化参与AMPK/mTOR信号通路促进宫颈癌细胞生长增殖和迁移。     

Anticancer Mechanism of Curcumin on Human Glioblastoma

Glioblastoma (GBM) is the most malignant brain tumor and accounts for most adult brain tumors. Current available treatment options for GBM are multimodal, which include surgical resection, radiation, and chemotherapy. Despite the significant advances in diagnostic and therapeutic approaches, GBM remains largely resistant to treatment, with a poor median survival rate between 12 and 18 months. With increasing drug resistance, the introduction of phytochemicals into current GBM treatment has become a potential strategy to combat GBM. Phytochemicals possess multifarious bioactivities with multitarget sites and comparatively marginal toxicity. Among them, curcumin is the most studied compound described as a potential anticancer agent due to its multi-targeted signaling/molecular pathways properties. Curcumin possesses the ability to modulate the core pathways involved in GBM cell proliferation, apoptosis, cell cycle arrest, autophagy, paraptosis, oxidative stress, and tumor cell motility. This review discusses curcumin’s anticancer mechanism through modulation of Rb, p53, MAPK, P13K/Akt, JAK/STAT, Shh, and NF-κB pathways, which are commonly involved and dysregulated in preclinical and clinical GBM models. In addition, limitation issues such as bioavailability, pharmacokinetics perspectives strategies, and clinical trials were discussed.     

Foxtail Millet Improves Blood Glucose Metabolism in Diabetic Rats through PI3K/AKT and NF-κB Signaling Pathways Mediated by Gut Microbiota

Foxtail millet (FM) is receiving ongoing increased attention due to its beneficial health effects, including the hypoglycemic effect. However, the underlying mechanisms of the hypoglycemic effect have been underexplored. In the present study, the hypoglycemic effect of FM supplementation was confirmed again in high-fat diet and streptozotocin-induced diabetic rats with significantly decreased fasting glucose (FG), glycated serum protein, and areas under the glucose tolerance test (p < 0.05). We employed 16S rRNA and liver RNA sequencing technologies to identify the target gut microbes and signaling pathways involved in the hypoglycemic effect of FM supplementation. The results showed that FM supplementation significantly increased the relative abundance of Lactobacillus and Ruminococcus_2, which were significantly negatively correlated with FG and 2-h glucose. FM supplementation significantly reversed the trends of gene expression in diabetic rats. Specifically, FM supplementation inhibited gluconeogenesis, stimulated glycolysis, and restored fatty acid synthesis through activation of the PI3K/AKT signaling pathway. FM also reduced inflammation through inhibition of the NF-κB signaling pathway. Spearman’s correlation analysis indicated a complicated set of interdependencies among the gut microbiota, signaling pathways, and metabolic parameters. Collectively, the above results suggest that the hypoglycemic effect of FM was at least partially mediated by the increased relative abundance of Lactobacillus, activation of the PI3K/AKT signaling pathway, and inhibition of the NF-κB signaling pathway.     

腹主动脉瘤腔内修复术后感染病原菌及Hippo信号通路基因改变

目的探讨腹主动脉瘤腔内修复术(EVAR)后医院感染病原特点及Hippo信号通路基因、白细胞介素-17(Interleukin-17,IL-17)、IL-23改变。方法选择天津医院血管外科2017年5月-2020年3月收治腹主动脉瘤EVAR术后医院感染患者49例作为感染组,选择同期医院进行EVAR术后未发生医院感染患者60例作为非感染组。采用实时荧光定量逆转录聚合酶链反应(RT-PCR)法检测Hippo通路基因yap、taz、mst1相对表达水平,采用酶联免疫吸附法检测其辅助型T细胞17(Th17)促炎细胞因子白细胞介素-17(IL-17)、IL-23水平。结果49例患者共发生肺部感染33例(67.35%),尿路感染10例(20.41%),手术切口感染4例(8.16%),移植物感染2例(4.08%);感染病原以革兰阳性菌为主,共25株,占51.02%,革兰阴性菌共22株,占44.90%,真菌2株,占4.08%;感染组yap、taz、mst1基因相对表达水平和IL-17、IL-23水平高于非感染组(P<0.05)。结论腹主动脉瘤EVAR术后医院感染以肺部感染、尿路感染为主,应及时予以针对性预防;Hippo通路参与了感染发生过程,通过Th17促炎途径诱导炎症反应,其机制仍有待研究。     

慢性乙型肝炎患者外周血SOCS3 mRNA的表达及其与Th17的关系

 目的 研究慢性乙型肝炎(CHB)患者外周血辅助性T细胞17(Th17)以及细胞因子信号转导抑制因子3(SOCS3) mRNA的表达状况,探索CHB患者SOCS3 mRNA表达与Th17的关系。方法 选取2021年2—8月于某院门诊就诊的30例CHB患者为研究对象,并选取同期正常体检者15例为对照组。采用流式细胞术(FCM)检测外周血Th17细胞频数,酶联免疫吸附试验(ELISA)检测血清细胞因子IL-17A和IL-23表达水平,实时荧光定量逆转录聚合酶链反应(qRT-PCR)法测定外周血SOCS3、维甲酸相关孤儿核受体γt (RORγt) mRNA表达水平,并比较两组患者的检测结果。结果 CHB患者外周血Th17细胞频数及其效应分子IL-17A、IL-23表达水平高于对照组(均P＜0.05),Th17细胞频数、IL-17A与HBV DNA水平之间存在正相关(r值分别为0.570、0.563,均P＜0.005)。CHB患者外周血SOCS3、RORγt mRNA表达水平高于对照组(均P＜0.05),两者与HBV DNA水平正相关(r值分别为0.662、0.561,均P＜0.05)。CHB患者SOCS3 mRNA与RORγt mRNA、Th17细胞频数、IL-17A之间存在正相关(r值分别为0.552、0.626、0.826,均P＜0.05)。结论 CHB患者外周血SOCS3 mRNA的异常高表达,可能通过调节RORγt mRNA的表达来影响Th17的分化及其效应分子的分泌。     

A novel adipokine WISP1 attenuates lipopolysaccharide-induced cell injury in 3T3-L1 adipocytes by regulating the PI3K/Akt pathway

BackgroundTo study the effect of WISP1 on lipopolysaccharide (LPS)-induced cell injury in 3T3-L1 adipocytes.MethodLentivirus was transiently transfected into log phase 3T3-L1 adipocytes, which were then treated with LPS at a concentration of 10 μg/mL for 24 h. The cells were divided into the following groups: group A (control, untreated cells); group B (LPS-treated cells); group C (GFP), cells transfected with lentivirus-containing GFP; group D (GFP+LPS), group C treated with LPS;group E (WISP1OE), cells transfected with lentivirus, group F (shNC+LPS), cells transfected with lentivirus-containing nshRNA treated with LPS; group G (shWISP1 +LPS), cells transfected with lentivirus-containing shRNA treated with LPS; group H (WISP1OE+LPS), group E treated with LPS; group I (WISP1OE+LPS+LY294002), group E treated with LPS followed by LY294002 for 24 h.ResultsWISP1 overexpression notably ameliorated cell apoptosis, accompanied with the increased expression of bcl-2, the decreased expressions of bax and cleaved-caspase-3, and promoted the release of inflammatory factors, such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in LPS-treated 3T3-L1 adipocytes. WISP1 knockdown exhibited the opposite results. In addition, WISP1 stimulated Akt phosphorylation and reduced nuclear translocation of Fork head box protein O3 (FoxO3a) in 3T3-L1 adipocytes treated by LPS. The inhibition of the PI3K/Akt signaling pathway diminished the protective effect of WISP1.ConclusionWISP1 prevents 3T3-L1 adipocytes from being injured by LPS by regulating the PI3K/Akt pathway.     

膜肾1号方对大鼠肾脏病理及PI3K/Akt/mTOR信号通路表达的影响

[目的] 观察膜肾1号方对膜性肾病大鼠肾脏病理的改善作用及其对自噬通路磷脂肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/雷帕霉素靶蛋白（mTOR）相关蛋白表达的影响。[方法] 将大鼠随机分为对照组、模型组、膜肾1号方高剂量组、中剂量组、低剂量组,盐酸贝那普利组。采用大鼠尾静脉注射阳离子化牛血清白蛋白（C-BSA）的方法建立MN大鼠模型,灌胃、取材。苏木精-伊红（HE ）染色法观察大鼠肾脏组织病理改变;免疫球蛋白G（IgG）免疫荧光染色观察大鼠IgG沉积;蛋白免疫印迹法（Western Blot）检测PI3K/Akt/mTOR 信号通路相关蛋白及自噬相关蛋白轻链3（LC3）表达。[结果] 药物干预后,膜肾1号方组大鼠24 h尿蛋白、三酰甘油、总胆固醇、低密度脂蛋白下降且低于模型组,并具有统计学差异（P＜0.05）。光镜下观察,HE染色示正常组肾组织整体结构基本正常,膜性肾病（MN）模型组肾小球毛细血管丛充血,系膜增生,基底膜出现增厚,部分肾小管细胞空泡变、组织内可见炎症细胞浸润,可见嗜复红蛋白及IgG沉积。经膜肾1号方和盐酸贝那普利干预后,大鼠肾脏病理学改变均有所减轻。各组大鼠IgG沉积显示,与对照组比较模型组IgG沉积明显,IgG荧光表达升高,差异具有统计学意义（P＜0.05）,盐酸贝那普利组和膜肾1号方组IgG荧光表达下降且低于模型组,具有统计学差异（P＜0.05）。Western Blot检测显示,药物干预后,盐酸贝那普利组和膜肾1号方组大鼠PI3K-Akt信号通路相关蛋白表达下降,LC3 表达增加,并具有统计学差异（P＜0.05）。[结论] 膜肾1号方可改善大鼠肾脏病理损伤,膜肾1号方干预后PI3K-Akt信号通路相关蛋白磷酸化磷酸肌醇3激酶（p-PI3K）,磷酸化Akt蛋白（p-Akt）,磷酸化雷帕霉素靶蛋白（p-mTOR）表达明显降低,自噬相关蛋白LC3表达升高,其分子机制与自噬信号通路的调控有关。     

基于代谢组学技术的黄芩-白芍对2型糖尿病模型小鼠的作用机制

目的：在肾脏代谢组学技术基础上,探讨黄芩-白芍药对对2型糖尿病（T2DM）的防治机制,并通过寻找内源性潜在生物标志物,寻找其相关代谢通路。方法：非糖尿病健康同窝出生仔畜(db/m小鼠）为空白组,同时选择血糖值高于11.1 mol/L的糖尿病瘦素受体基因缺陷（db/db）小鼠进行分组,分为模型组、黄芩-白芍组（5.625 g/kg）、阳性药组（二甲双胍250 mg/kg）,给药体积为7.5 mL/kg,1次/d,连续8周,并采用超高效液相色谱与串联四级杆飞行时间质谱仪联用技术(UPLC-Q-TOF-MS/MS)法进行小鼠肾脏代谢组学分析,探寻潜在的生物标志物,联合人类代谢组学数据库（HMDB）和京都基因与基因组百科全书数据库（KEGG）找出相关代谢通路。结果：黄芩-白芍组可改善T2DM模型小鼠代谢轨迹的偏离,同时,寻找出与DN模型及给予黄芩汤干预后具有共同差异的生物标志物14个,涉及8条相关代谢通路,应用相关代谢通路分析方法发现,影响最为广泛的代谢通路可能是甘油磷脂代谢、嘌呤代谢和嘧啶代谢。结论：黄芩-白芍药对治疗T2DM的作用机制可能与其通过调节脂质代谢、能量代谢等多条代谢通路,改善内源性代谢物的水平,恢复机体正常代谢活动有关。     

川陈皮素通过激活SIRT1/FOXO3a信号来减轻肾缺血再灌注损伤

目的：本研究旨在探究川陈皮素（NOB）保护小鼠肾缺血再灌注损伤（RIRI）的可能分子机制。方法：将Balb/c小鼠分为5组（n=6）：假手术组、模型组、NOB组（50 mg/kg）、组蛋白去乙酰化酶沉默信息调节因子1（SIRT1）抑制剂EX527组（5 mg/kg）、NOB+EX527组（50 mg/kg的NOB+5 mg/kg EX527）。在建模前24 h对小鼠进行药物处理。通过阻断小鼠左肾动静脉血流建立RIRI模型。建模24 h后检测肾组织中氧化应激标志物含量。通过苏木精-伊红（HE）染色和天狼星红染色评价肾组织病变和纤维化。通过TUNEL染色检测肾细胞凋亡。通过蛋白质免疫印迹法检测肾组织中SIRT1、叉头框蛋白O3a（FOXO3a）、细胞凋亡和自噬相关蛋白表达。通过实时荧光定量PCR检测肾组织中SIRT1和FOXO3a mRNA的水平。结果：与模型组比较,NOB组小鼠肾脏病变程度减轻,肾脏纤维化面积降低（均P<0.05）。与模型组比较,NOB组肾组织抗氧化作用升高（P<0.05）。与模型组比较,NOB组肾组织中细胞凋亡减少（P<0.05）。与模型组比较,NOB组肾组织中SIRT1和FOXO3a的mRNA和蛋白相对表达量升高,微管相关蛋白轻链(LC)3Ⅱ/LC3Ⅰ蛋白相对表达量升高,而p62降低（均P<0.05）。此外,EX527逆转了NOB对肾脏的保护作用（P<0.05）。结论：NOB通过激活SIRT-1/FOXO3a信号通路介导的自噬来减轻RIRI。