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。     

铁皮石斛水提物对4T1乳腺癌荷瘤小鼠的抑瘤及免疫调节作用研究

目的 研究铁皮石斛Dendrobium officinale水提物对4T1乳腺癌荷瘤模型小鼠的抑瘤及免疫调节作用,并进一步探讨其作用机制。方法 采用BALB/c雌性小鼠构建4T1乳腺癌荷瘤小鼠模型后,随机分为模型组、盐酸表柔比星组（8 mL/kg）和铁皮石斛水提物低、中、高剂量（0.5、1.0、1.5 g/kg）组,另选10只正常BALB/c雌性小鼠作为对照组,给予相应药物进行干预,观察小鼠生存状态,每3天测量肿瘤直径与短径,连续给药28 d后处死小鼠,称定肿瘤质量,计算抑瘤率;采用小动物活体成像仪和转移灶计数观察肺部肿瘤转移情况;称定小鼠脾脏、胸腺质量,计算脏器系数;苏木素-伊红（HE）染色观察肿瘤、肺部以及脾脏形态学变化;血液分析仪检测荷瘤小鼠外周血中白细胞数目和淋巴细胞占比;ELISA法检测血清中白细胞介素-2(interleukin-2,IL-2)、γ干扰素（interferon-γ,IFN-γ）和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;流式细胞术检测外周血中T淋巴细胞比例;脾淋巴细胞增殖试验检测脾脏中T淋巴细胞、B淋巴细胞增殖能力;...     

TWIST2通过调控FGF21介导的AMPK/mTOR通路诱导卵巢癌细胞氧化应激和凋亡

目的探究TWIST2对卵巢癌细胞氧化应激和凋亡的作用及相关机制。方法收集2018年6月至2019年10月在山东省妇幼保健院进行治疗的30例卵巢癌患者的癌组织和癌旁组织,将pcDNA3.1-TWIST2质粒(TWIST2组)和pcDNA3.1阴性对照质粒(阴性对照组)分别转染卵巢癌细胞(SK-OV-3和HO-8910),未转染质粒组为空白对照组。通过实时荧光定量PCR(qRT-PCR)和免疫印迹(Western blot)实验检测组织和细胞中TWIST2 mRNA和蛋白质的表达量;细胞计数试剂盒CCK-8检测TWIST2组、阴性对照组和空白对照组的细胞活力;Western blot实验检测3组的凋亡相关蛋白caspase-3、Bax和Bcl-2,AMPK/mTOR通路相关蛋白FGF21、p-AMPK和p-mTOR的蛋白表达水平;试剂盒检测3组活性氧(ROS)基因、ATP的水平和超氧化物歧化酶(SOD)的酶活性。通过检测不同处理组的细胞凋亡率和ROS水平,研究TWIST2与FGF21的相互作用。结果TWIST2在卵巢癌组织和卵巢癌细胞中的表达量显著低于癌旁组织和正常卵巢上皮细胞的表达量(P<0.05)。与空白对照组和阴性对照组相比,TWIST2组的细胞活力显著降低,细胞凋亡率显著提高(P<0.05);ROS的水平显著升高,SOD酶活力、ATP水平、NAD~+/NADH水平显著降低(P<0.05)。TWIST2可以抑制FGF21、p-AMPK、p-mTOR蛋白的表达(P<0.05);在TWIST2组中加入FGF21或AMPK激动剂时,p-AMPK、p-mTOR蛋白表达水平显著升高,卵巢癌细胞的凋亡率和ROS水平显著降低(P<0.05),TWIST2与FGF21的表达量呈负相关(r=-0.6254,P<0.01)。结论TWIST2能够促进卵巢癌细胞氧化应激和凋亡,其作用机制是通过调控FGF21介导的AMPK/mTOR通路来实现的。     

槐定碱通过PI3K/Akt/mTOR信号通路诱导自噬抑制胰腺癌细胞增殖

目的 探讨槐定碱对胰腺癌细胞增殖及自噬的影响,并分析其机制。方法 MTT法分析Sw1990细胞增殖, MDC染色法检测细胞自噬水平,western blot检测细胞自噬相关蛋白、PI3K/Akt/mTOR信号通路相关蛋白表达水平,运用自噬抑制剂（3 - MA）研究自噬对细胞增殖的影响;裸鼠成瘤实验检测体内胰腺癌细胞增殖情况,并分析瘤组织中LC3 II、p - mTOR蛋白水平。结果 槐定碱抑制胰腺癌Sw1990细胞的增长,促进自噬小泡的形成,上调LC3 II/ LC3 I、Beclin - 1水平,下调p - PI3K、p - AKT、p - mTOR水平（P＜0.05）;与槐定碱40 μmol/L组比较,槐定碱40 μmol/L + 3 - MA 5 μmol/L组细胞抑制率升高,LC3 II/ LC3 I降低,p - mTOR蛋白水平升高（P＜0.05）; 40 mg/kg槐定碱下调裸鼠瘤体体积、瘤体质量,上调LC3 II/ LC3 I水平,下调p - mTOR蛋白水平（P＜0.05）。结论 槐定碱能抑制Sw1990细胞增殖,与调控PI3K/Akt/mTOR信号通路影响自噬有关。     

mTOR信号通路在孤独症谱系障碍中的研究进展

哺乳动物雷帕霉素靶蛋白（mTOR）为细胞内信号通路转导分子,调节很多重要的生理过程。mTOR信号通路在神经系统的突触可塑性、信息传递和加工、神经调控等方面发挥重要作用。mTOR信号通路失调被认为与孤独症谱系障碍（ASD）的发病机制有关,mTOR抑制剂可以改善ASD的症状。本文对mTOR信号通路在ASD发病机制中的作用进行综述,以期为ASD的靶向治疗提供理论依据。     

The next era of treatment for hormone receptor-positive,HER2-negative advanced breast cancer: Triplet combination-based endocrine therapies

Until recently, the standard of care for hormone receptor-positive (HR+) breast cancer was single-agent endocrine therapy, which aims to prevent estrogen receptor signaling. This therapeutic strategy has extended survival without the toxicity associated with chemotherapy, but primary endocrine therapy resistance is common, and secondary resistance develops over time. Adjunct downstream inhibition of the cyclin-dependent kinase (CDK)4/6 pathway, intended to delay and prevent endocrine therapy resistance, has further extended progression-free survival in patients receiving endocrine therapy; however, resistance still eventually develops in these patients. Addition of phosphatidylinositol-3 kinase (PI3K) or mammalian target of rapamycin (mTOR) inhibitors to combined CDK4/6 and endocrine inhibitor regimens may help prolong CDK4/6 inhibitor sensitivity. Early trials combining CDK4/6 inhibitors, PI3K or mTOR inhibitors, and endocrine therapy have shown encouraging signs of clinical activity. However, further research is needed to help understand the extent of treatment benefit from triplet therapy and where this strategy will fit in the treatment sequence for patients with HR+ breast cancer.     

Optimal treatment of poor-risk renal cell carcinoma patients with mTOR inhibitors and anti-VEGFR agents

Poor-risk metastatic renal cell carcinoma (RCC) includes a subgroup of patients with unfavorable prognosis, according to both the Motzer and Heng criteria. Overall, owing to the poor prognosis of these patients, the approach is still a challenge for the first and subsequent lines of treatment, particularly for rare histologies other than clear cell renal cell carcinoma. In this review, we investigated the present treatment option of poor-risk metastatic RCC. Areas covered are data with first and further line of therapy with mTOR inhibitors and other agents but without cytoreductive nephrectomy or rare histologies. The current data on systemic therapy in poor-risk metastatic RCC maintain temsirolimus as the preferred first-line therapy. New agents targeting immune checkpoints are being developed in clinical trials.     

Identification of biomarkers for essential hypertension based on metabolomics

AimEssential hypertension (EH) is one of the most important public health problems worldwide. However, the pathogenesis of EH is unclear and early diagnostic methods are lacking. Metabolomics demonstrates great potential for biomarker discovery and the mechanistic exploration of metabolic diseases.Data synthesisThis review included human and animal metabolomics studies related to EH in the PubMed and Web of Science databases between February 1996 and May 2020. The study designs, EH standards, and reported metabolic biomarkers were systematically examined and compared. The pathway analysis was conducted through the online software MetaboAnalyst 4.0.Twenty-two human studies and fifteen animal studies were included in this systematic review. There were many frequently reported biomarkers with consistent trends (e.g., pyruvate, lactic acid, valine, and tryptophan) in human and animal studies, and thus had potential as biomarkers of EH. In addition, several shared metabolic pathways, including alanine, aspartate, and glutamate metabolism, aminoacyl-tRNA biosynthesis, and arginine biosynthesis, were identified in human and animal metabolomics studies. These biomarkers and pathways, closely related to insulin resistance, the inflammatory state, and impaired nitric oxide production, were demonstrated to contribute to EH development.ConclusionsThis study summarized valuable metabolic biomarkers and pathways that could offer opportunities for the early diagnosis or prediction of EH and the discovery of the metabolic mechanisms of EH.