Resveratrol attenuates intestinal injury in irradiated rats via PI3K/Akt/mTOR signaling pathway

Radiation‐induced enteritis is one of the greatest challenges in radiotherapy. The current study was designed to evaluate the ameliorative effect of resveratrol, which exhibits anti‐inflammatory property, against radiation‐induced intestinal injury in rats and to explore the underlying mechanism. Rats were exposed to a single dose of 5 Gy. Resveratrol (20 mg/kg/day) was orally administered to irradiated rats over 3 weeks. Results showed that resveratrol ameliorated the intestinal oxidative stress parameters; malondialdehyde (MDA) content, glutathione (GSH) level, and catalase (CAT) activity compared to irradiated group. Furthermore, resveratrol reduced the contents of inflammatory cytokines; tumor necrosis factor α (TNF‐α), nuclear factor‐kappa (NF‐κB), and interleukin 1β (IL‐1β) in intestine. Western blotting analysis revealed that resveratrol down‐regulated the proteins expression of phosphoinositide 3‐kinases (PI3K), protein kinase B (Akt) as well as the mammalian target of rapamycin (mTOR) in intestinal tissues of irradiated rats and thus reduced the inflammatory mediator production. These results were confirmed by histopathological investigation. In conclusion, resveratrol attenuated intestinal inflammation following irradiation via modulating PI3K/Akt/mTOR pathway and thereby could be a promising adjuvant in radiotherapy.     

PI3K/Akt/mTOR信号通路与肿瘤

在近年来的肿瘤治疗中,靶向生物治疗逐渐成为研究的热点。该文就磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白[phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt)/the mammalian target of Rapamycin(mTOR),PI3K/Akt/mTOR]信号通路予以综述,重点包括PI3K/Akt/mTOR信号转导在肿瘤机制中作用以及肿瘤治疗过程中耐药性方面的关系等。     

The PI3K/Akt/mTOR pathway: A potential pharmacological target in COVID-19

Coronavirus disease 2019 (COVID-19) has emerged as a serious threat to global health. The disregulation of the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) cell signaling pathway observed in patients with COVID-19 has attracted attention for the possible use of specific inhibitors of this pathway for the treatment of the disease. Here, we review emerging data on the involvement of the PI3K/Akt/mTOR pathway in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and the clinical studies investigating its tailored inhibition in COVID-19. Current in silico, in vitro, and in vivo data convergently support a role for the PI3K/Akt/mTOR pathway in COVID-19 and suggest the use of specific inhibitors of this pathway that, by a combined mechanism entailing downregulation of excessive inflammatory reactions, cell protection, and antiviral effects, could ameliorate the course of COVID-19.     

补骨脂素对绝经后大鼠骨质疏松及PI3K/Akt/mTOR信号通路的影响

目的:研究补骨脂素对绝经后大鼠骨质疏松及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的影响。方法:将60只健康雌性SD大鼠随机分为正常组、模型组、阳性对照组(0.09 mg/kg雌二醇)和补骨脂素低、中、高剂量组(22、44、88 mg/kg),每组10只。除正常组外,其余各组大鼠均采用卵巢摘除去势法建立绝经后骨质疏松模型。术后正常饲养2个月,正常组和模型组大鼠灌胃等体积生理盐水,各药物组大鼠灌胃相应药液;灌胃体积均为0.005 mL/g,每天1次,连续98天。末次给药24 h后,测定大鼠右侧下肢股骨和椎骨的骨密度,血清中钙离子、骨钙素、Ⅰ型前胶原N端前肽(P1NP)含量和骨形态发生蛋白2(BMP2)、血管内皮生长因子(VEGF)水平,以及股骨组织中PI3K、Akt、mTOR mRNA及蛋白的表达水平。结果:与正常组比较,模型组大鼠股骨和椎骨的骨密度以及血清中钙离子、骨钙素、P1NP含量和BMP2、VEGF水平均显著降低,PI3K、Akt、mTOR mRNA及蛋白的表达水平均显著升高(P<0.05或P<0.01)。与模型组比较,补骨脂素中、高剂量组和阳性对照组大鼠股骨和椎骨的骨密度以及血清中钙离子、骨钙素、P1NP含量和BMP2(补骨脂素中剂量组除外)、VEGF(补骨脂素中剂量组除外)水平均显著升高,各药物组PI3K、Akt、m TOR mRNA(补骨脂素低剂量组除外)及蛋白表达水平均显著降低(P<0.05或P<0.01),且高剂量组股骨骨密度和钙离子、BMP2水平以及PI3K蛋白表达水平均显著高于阳性对照组(P<0.05),mTOR mRNA表达水平显著低于阳性对照组(P<0.05)。结论:补骨脂素可改善绝经后大鼠的骨质疏松,其机制可能与抑制PI3K/Akt/mTOR信号通路有关。     

Update on Staphylococcal Superantigen-Induced Signaling Pathways and Therapeutic Interventions

Staphylococcal enterotoxin B (SEB) and related bacterial toxins cause diseases in humans and laboratory animals ranging from food poisoning, acute lung injury to toxic shock. These superantigens bind directly to the major histocompatibility complex class II molecules on antigen-presenting cells and specific Vβ regions of T-cell receptors (TCR), resulting in rapid hyper-activation of the host immune system. In addition to TCR and co-stimulatory signals, proinflammatory mediators activate signaling pathways culminating in cell-stress response, activation of NFκB and mammalian target of rapamycin (mTOR). This article presents a concise review of superantigen-activated signaling pathways and focuses on the therapeutic challenges against bacterial superantigens.     

PI3K/Akt/mTOR信号传导通路与前列腺癌关系的研究进展

前列腺癌是威胁中老年男性健康的常见肿瘤,成为男性癌症死因的第二位。 PI3K/Akt/mTOR信号通路能够通过维持细胞生存、抑制细胞凋亡、促进细胞周期运行及血管生成等促进前列腺癌病程发展。本文综合国内外文献,阐述PI3K/Akt/mTOR信号通路在前列腺癌发生发展中的作用以及和通路相关的药物治疗进展。     

BAFF/BAFF-R通过PI3K/Akt/mTOR信号转导通路调节B淋巴细胞功能在类风湿关节炎发病机制中的意义

B淋巴细胞活化和生存在类风湿关节炎(RA)病程中起关键作用。肿瘤坏死因子家族B细胞活化因子(BAFF)是维持B细胞功能的重要细胞因子。BAFF与其受体BAFF-R结合(BAFF/BAFF-R),能够激活PI3K/Akt/mTOR信号通路,调节B淋巴细胞的增殖、存活和活化。该文就B淋巴细胞、BAFF/BAFF-R以及PI3K/Akt/mTOR信号通路参与RA的发病机制加以综述。     

ROS-dependent inhibition of the PI3K/Akt/mTOR signaling is required for Oroxylin A to exert anti-inflammatory activity in liver fibrosis

More and more evidence showed that autophagy is an inflammation-related defense mechanism against a variety of diseases including liver fibrosis. However, the essential mechanisms remain poorly understood. In this study, we sought to elucidate the impact of Oroxylin A on autophagy and further to identify the potential mechanism of its anti-inflammatory activity. We found that Oroxylin A played a critical role in controlling inflammation in murine liver fibrosis. Moreover, Oroxylin A could inhibit the secretion of pro-inflammatory cytokines in activated hepatic stellate cell (HSCs). We previously reported that Oroxylin A can induce autophagy to alleviate the pathological changes of liver fibrosis and the activation of HSC. Here we further revealed that the inhibition of the PI3K/Akt/mTOR signaling was required for Oroxylin A to induce autophagy activation, which may be the underlying mechanism of the anti-inflammatory activity of Oroxylin A. Interestingly, mTOR overexpression completely impaired the Oroxylin A-mediated autophagy activation, and in turn, damaged the anti-inflammatory activity. Importantly, Oroxylin A inhibited PI3K/Akt/mTOR signaling by scavenging reactive oxygen species (ROS). ROS accumulation by buthionine sulfoximine (BSO) could abrogate the Oroxylin A-mediated ROS elimination, the inhibition of PI3K/Akt/mTOR signaling, and anti-inflammatory activities. Overall, our results provided reliable evidence for the molecular mechanism of Oroxylin A-mediated anti-fibrosis activity, and also identified a new target for drug therapy of liver fibrosis.     

姜黄素通过抑制PI3K/Akt/mTOR通路增强自噬保护帕金森病细胞模型的研究

目的 观察姜黄素对帕金森病（Parkinson’s disease,PD）细胞模型中磷脂酰肌醇-3-激酶（phosphatidylinositol-3-kinase,PI3K）/蛋白激酶B （protein kinase B,Akt）/哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin,mTOR）通路的影响,并探讨其是否通过抑制此通路来增强自噬而发挥神经保护作用。方法 采用1-甲基-4-苯基-四氢吡啶离子处理人神经母细胞瘤SH-SY5Y细胞建立PD细胞模型,设立对照组、模型组、姜黄素组、姜黄素+胰岛素样生长因子1（insulin-like growth factor 1,IGF-1）组、姜黄素+LY294002组,分别应用姜黄素、姜黄素联合PI3K通路激活剂IGF-1、姜黄素联合PI3K通路抑制剂LY294002进行干预处理。各组细胞在药物处理24 h后分别于光学显微镜下观察细胞形态,细胞计数试剂盒检测细胞活力,酪氨酸羟化酶免疫荧光染色观察多巴胺（dopamine,DA）能神经元存活数目,Western blotting检测PI3K、磷酸化-Akt （phospho-Akt,p-Akt）、磷酸化-mTOR （phospho-mTOR,p-mTOR）、α-突触核蛋白（α-synuclein,α-Syn）和微管相关蛋白1轻链3B （microtubule-associated protein 1 light chain 3 beta,LC3B）的蛋白表达。结果 与模型组比较,姜黄素组细胞皱缩、空泡变性的状态得到改善,细胞存活率提高（P<0.05）,DA能神经元存活数目增加（P<0.01）,LC3B-II/LC3B-I比值增加（P<0.05）,α-Syn蛋白表达减少（P<0.01）,p-Akt、p-mTOR的蛋白表达显著下调（P<0.01）。添加PI3K通路激活剂IGF-1,姜黄素的上述作用基本被抵消;添加PI3K通路抑制剂LY294002后,与单一姜黄素干预比较,虽然LC3B-II/LC3B-I比值进一步增加（P<0.05）,但α-Syn蛋白表达增加（P<0.05）,DA能神经元存活数减少（P<0.01）。结论 在PD细胞模型中姜黄素可抑制PI3K/Akt/mTOR信号通路的活化,从而增强细胞自噬功能,继而促进α-Syn的清除是其发挥神经保护作用的主要机制之一。     

丹皮酚与顺铂联用对人骨肉瘤细胞MG-63增殖、凋亡及PI3K/Akt/mTOR信号通路的影响

目的:研究丹皮酚与顺铂联用对人骨肉瘤细胞MG-63增殖、凋亡的影响及可能的作用机制.方法:取对数生长期MG-63细胞,分为空白对照组、顺铂组(4μmol/L)、丹皮酚组(50 mg/L)和低、中、高浓度联用组(50、100、200 mg/L丹皮酚+4μmol/L顺铂).采用CCK-8法检测经药物作用24、48、72 h...     

Urolithin A suppresses tumor progression and induces autophagy in gastric cancer via the PI3K/Akt/mTOR pathway

Urolithin A (UA) is a microbial metabolite of natural polyphenols ellagitannins and ellagic acid with well-established antitumor properties against various malignancies. However, the exact role of UA in gastric cancer (GC) progression remains largely unclear. In the present study, we investigated the effects and potential mechanisms of UA in GC in vitro and in vivo. Our results revealed that UA could suppress GC cell proliferation, inhibit migration and invasion, promote apoptosis, and induce autophagy via the phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin pathway in vitro. The autophagy inhibitors 3-methyladenine and chloroquine augmented the inhibitory effect of UA on proliferation and promoted apoptosis, implying that UA mediated the cytoprotective role of autophagy. Meanwhile, the in vivo experiments showed that UA effectively suppressed tumor growth, enhanced the therapeutic effects, and alleviated chemotherapy toxicity in xenograft models. Overall, these findings offer novel insights into the role of UA in tumor therapy and suggest that UA may possess potential therapeutic applications for GC.     

Investigational PI3K/AKT/mTOR inhibitors in development for endometrial cancer

Introduction: Endometrial cancer (EC) is the most common neoplasm of the female genital tract in developed countries. Despite the progress in early detection and treatment, a significant number of cases of advanced ECs are still diagnosed. These patients have few treatment options and a poor prognosis. Our understanding of EC pathogenesis and progression has been enhanced by recent genomic studies. Among the relevant biological pathways, phosphatidylinositol 3-kinase/AKT (PIK3/AKT)-mammalian target of rapamycin (mTOR) signaling is frequently upregulated in this cancer.

Areas covered: This review covers investigational EC therapeutics acting on the PI3K/AKT/mTOR pathway. The authors review the results of clinical studies and highlight ongoing trials.

Expert opinion: Several new agents are under evaluation for treating patients with metastatic, recurrent, and persistent EC. Clinical trials investigating PI3K/AKT/mTOR inhibitors have yielded controversial results. In the near future, new studies with dual inhibitors or multi-pathways inhibitors as mono or combination therapies with conventional chemotherapy (CT) or other targeted drugs may provide more promising data. Moreover, the evaluation of new serum and histological biomarkers is an attractive strategy for patient selection.     

PI3K/Akt/mTOR通路在炎症相关疾病中分子机制研究进展

炎症是一种机体应对感染、组织损伤或者细胞应激的反应,并且可以通过修复机制恢复组织功能。炎症发生时会引起多条信号通路的激活,包括核转录因子-κB（NF-κB）通路、Janus激酶/信号转导与转录激活子（JAK/STAT）通路、丝裂原活化蛋白激酶（MAPK）通路以及磷脂酰肌醇-3-激酶/蛋白激酶B /雷帕霉素靶蛋白（PI3K/Akt/mTOR）通路等。本文综述了近年来磷脂酰肌醇-3-激酶/蛋白激酶B /雷帕霉素靶蛋白通路在炎症相关疾病中的分子作用机制,为研发以磷脂酰肌醇-3-激酶/蛋白激酶B /雷帕霉素靶蛋白为靶点的药物提供理论依据。     

大黄素下调ILK/PI3K/Akt信号通路抑制结肠癌CACO-2细胞增殖的机制研究

目的探讨大黄素对人结肠癌CACO-2细胞生长、增殖的影响,并探讨其可能的机制。方法体外培养人结肠癌CACO-2细胞,用不同浓度的大黄素(10,20,40,80μmol·L^-1)处理不同的时间(24,48,72 h)。采用MTT检测结肠癌CACO-2细胞的增殖活力,流式细胞术细胞周期的变化;JC-1检测线粒体膜电位的变化;Western blot检测细胞内ILK(整合素蛋白激酶)、p-PI3K和p-Akt的蛋白水平的表达。结果与空白对照组和DMSO对照组比较,大黄素可显著地抑制CACO-2细胞的增殖,并且存在显著的剂量-时间依赖性(P<0.01);流式细胞术结果表明,大黄素可使细胞的增殖阻滞于G0/G1期;JC-1染色结果显示大黄素可显著降低线粒体膜电位(P<0.01);Western blot结果显示大黄素能够显著降低细胞内ILK、p-PI3K和p-Akt蛋白的表达(P<0.01),且具有浓度-时间依赖性(P<0.01)。结论大黄素对人结肠癌CACO-2细胞表现成明显的抗癌作用,其机制与大黄素抑制ILK/PI3K/Akt信号通路进而抑制癌细胞增殖、诱导细胞周期阻滞于G0/G1期、促进凋亡有关。     

当归挥发油通过抑制PI3K/Akt/mTOR信号转导通路影响人结肠癌LOVO细胞自噬研究

目的 研究当归挥发油(volatile oil of Angelicae Sinensis Radix,VOAS)对人结直肠癌LOVO细胞自噬的影响及其分子机制。方法 用不同浓度的VOAS (6.25,12.5,25,50和100 μg·mL^-1)作用于人结肠癌LOVO细胞48 h,CCK8比色法检测VOAS对LOVO细胞增殖的抑制作用。将VOAS 50 μg·mL^-1作用于LOVO细胞,AO荧光染色和透射电镜观察细胞自噬情况;Western blotting观察VOAS对自噬相关蛋白LC3B、Beclin-1、Atg5及PI3K/Akt/mTOR信号转导通路的影响。结果 6.25~100 μg·mL^-1的VOAS能够抑制LOVO细胞的增殖,具有浓度依赖性;50 μg·mL^-1的VOAS能够促进LOVO细胞的自噬,上调自噬相关蛋白LC3B-Ⅱ、Beclin-1及Atg5的表达,同时下调PI3K、p-Akt、p-mTOR蛋白的表达。结论 VOAS能够促进人结肠癌LOVO细胞自噬,其机制可能与抑制PI3K/Akt/mTOR信号转导通路有关。     

紫草素抑制PI3K/Akt/mTOR信号通路诱导人结肠癌SW480细胞凋亡和自噬的作用研究

目的 探讨紫草素对人结肠癌SW480细胞凋亡和自噬的影响及其机制。方法 取对数生长期人结肠癌SW480细胞,设对照组（DMSO）、紫草素（0.3、0.5、0.7 μg/mL）和LY294002（PI3K特异性抑制剂,5 μg/mL）组。药物干预48 h后,四甲基偶氮唑盐（MTT）法检测SW480细胞增殖抑制率,Annexin V-FITC流式细胞术分析细胞凋亡状况,蛋白免疫印迹法（Western blotting）检测p-PI3K、p-Akt、p-mTOR、Caspase-3、cleaved Caspase-3、Bcl-2、Bax、LC3蛋白表达并计算Bax/Bcl-2和LC3-II/LC3-I值。结果 与对照组比较,经紫草素0.3、0.5、0.7 μg/mL或LY294002 5 μg/mL干预能够显著提高人结肠癌SW480细胞增殖抑制率和凋亡率（P<0.01）;经紫草素0.5、0.7 μg/mL或LY294002 5 μg/mL干预能够显著下调p-PI3K、p-Akt、p-mTOR、Bcl-2蛋白表达,并上调Caspase-3、cleaved Caspase-3、Bax蛋白表达（P<0.05、0.01）,提高Bax/Bcl-2和LC3-II/LC3-I值（P<0.01）。与LY294002组比较,经紫草素0.7 μg/mL干预能够显著提高SW480细胞增殖抑制率和凋亡率（P<0.05、0.01）,下调p-PI3K、p-Akt、p-mTOR蛋白表达并上调Caspase-3、cleaved Caspase-3、Bax蛋白表达（P<0.05、0.01）,提高Bax/Bcl-2和LC3-II/LC3-I值（P<0.01）。结论 紫草素能够促进人结肠癌SW480细胞凋亡和自噬,作用机制可能与抑制PI3K/Akt/mTOR信号通路活化有关。     

丹参酮IIA通过调控PI3K/Akt/mTOR信号通路对食管癌细胞放疗敏感性的影响

目的 研究丹参酮II_A对人食管癌细胞放疗敏感性的影响，并基于磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白（PI3K/Akt/mTOR）信号通路探讨其潜在机制。方法 以人食管癌Eca-109细胞为受试细胞，分别采用不同浓度丹参酮II_A和不同放射剂量处理Eca-109细胞，48 h后MTT法检测细胞增殖活力，计算丹参酮II_A半数抑制浓度（IC₅₀）和放射的IC₅₀，分别作为后续实验丹参酮II_A浓度和放射剂量。取对数生长期Eca-109细胞，设对照组、丹参酮II_A组、放射组、丹参酮II_A＋放射组、丹参酮II_A＋放射＋PI3K抑制剂（LY294002）组。通过平板克隆实验、MTT法、流式细胞术、荧光质粒转染法检测细胞克隆形成能力、增殖活力、凋亡率和自噬状况，RT-PCR、Western blotting法检测细胞中PI3K/Akt/mTOR信号通路相关mRNA和蛋白表达。结果 丹参酮Ⅱ_A和放射对人食管癌Eca-109细胞增殖活力的抑制作用均呈现剂量相关性，丹参酮II_A IC₅₀为8.75 mmol/L，放射量IC₅₀为4.63 Gy。与对照组相比，丹参酮II_A组、放射组、丹参酮II_A＋放射组、丹参酮II_A＋放射＋LY294002组细胞克隆形成能力和增殖活力明显降低，凋亡率和自噬体数量明显升高（P＜0.05）；PI3K、Akt、mTOR mRNA和蛋白表达量明显降低（P＜0.05）；Bcl-2、Bax、cleaved Caspase-3、LC3-I、LC3-II蛋白表达量及Bax/Bcl-2、Cleaved Caspase-3/Caspase-3、LC3-II/LC3-I明显升高（P＜0.05）。与丹参酮II_A组或放射组相比，丹参酮II_A＋放射组和丹参酮II_A＋放射＋LY294002组对各检测指标的调控作用明显增强（P＜0.05）。与丹参酮II_A＋放射组相比，丹参酮II_A＋放射＋LY294002组对各指标的调控作用明显增强（P＜0.05）。结论 丹参酮II_A可能通过下调PI3K/Akt/mTOR信号通路，抑制细胞增殖并促进其凋亡与自噬，进而增强人食管癌细胞放疗敏感性。     

基于ROS/PI3K/Akt/mTOR信号通路探讨玫瑰花甲醇提取物对前列腺癌PC-3细胞凋亡的影响

目的：探讨玫瑰花甲醇提取物（RE）对前列腺癌PC-3细胞增殖和凋亡的影响及潜在作用机制。方法：采用CCK8法检测不同浓度RE对PC-3细胞增殖的影响;采用Hoechst染色法和Annexin V/PI双染法检测细胞凋亡;DCFH-DA检测细胞ROS水平;Western blot法检测Bax、Bcl-2、cleaved caspase-3、Cyt-C、PI3K、p-PI3K、Akt、p-Akt、mTOR、p-mTOR的蛋白表达。结果：RE呈浓度依赖性抑制前列腺癌PC-3细胞增殖,24 h和48 h后的IC₅₀值分别为31.30 mg·mL^-1和16.76 mg·mL^-1;RE能显著诱导PC-3细胞凋亡（P<0.05,P<0.01）,且呈现浓度依赖性;RE可显著提高PC-3细胞ROS水平（P<0.05,P<0.01）,且具有浓度依赖性;RE能显著上调PC-3细胞Bax、cleaved caspase-3、Cyt-C的蛋白表达及Bax/Bcl-2的比值（P<0.05,P<0.01）,显著下调Bcl-2、p-PI3K、p-Akt、p-mTOR的蛋白表达（P<0.05,P<0.01）,以上均呈现浓度依赖性。结论：本研究表明RE在体外有明显抑制前列腺癌细胞PC-3的增殖作用,并可能通过调控ROS/PI3K/Akt/mTOR信号通路诱导其凋亡,研究结果可为玫瑰花及其组方临床治疗前列腺癌提供实验依据。     

红参对卵巢储备功能减退大鼠卵巢储备功能和卵巢组织磷脂酰肌醇 3-激酶/蛋白激酶 B/哺乳动物雷帕霉素靶蛋白信号通路的影响

目的探讨红参对卵巢储备功能减退(DOR)大鼠卵巢功能的改善作用以及潜在机制。方法 2022年 8—11月进行 DOR大鼠模型建立实验。将 48只 SD雌性大鼠分为空白组、模型组、模型+雌二醇组以及模型+红参组,每组 12只。采用腹腔注射去氧乙烯基环己烯(VCD)225 mg/kg建立 DOR大鼠模型。模型组与空白组均给予 0.9%氯化钠溶液灌胃,模型组+雌二醇组给予 0.15 mg/kg戊酸雌二醇溶液灌胃,模型+红参组给予 5 g/kg红参混悬液灌胃,1次/天,共 30 d。 30 d后比较各组大鼠治疗前后体质量变化;观察卵巢组织病理切片变化;检测血清促卵泡生成素(FSH)、促黄体生成素(LH)、雌二醇、促性腺激素释放激素(GnRH)和抗缪勒管激素(AMH)激素水平变化;蛋白质印迹法检测卵巢组织中磷脂酰肌醇 3-激酶 p85α(PI3K p85α)、蛋白激酶 B(Akt)和哺乳动物雷帕霉素靶蛋白(mTOR)蛋白表达量和磷酸化水平。结果用药后,模型+雌二醇组大鼠体质量差值为(15.0±0.7)g高于模型组的(10.9±1.3)g,体质量明显增加(P<0.05)。模型+红参组大鼠体质量差值为(14.9±0.4)g,高于模型组的(10.9±1.3)g(P<0.05),但与模型+雌二醇组的(15.0±0.7)g比较差异无统计学意义(P>0.05)。病理切片显示,模型+雌二醇组和模型+红参组卵泡数量多于模型组,卵巢结构较为清晰,颗粒结构更为紧凑,黄体数量较模型组增多;ELISA实验显示,用药后,模型+红参组和模型+雌二醇组 AMH、雌二醇和 GnRH明显升高,FSH、LH明显降低(P<0.05)其中模型+红参组激素水平恢复更明显(P<0.05)。蛋白质印迹法显示,与模型组比较,用药后,模型+红参组和模型+雌二醇组,p-PI3K p85α、p-Akt和 pmTOR均显著下降(P<0.05);与雌二醇组比较,模型+红参组 p-PI3K p85α、p-Akt、p-mTOR水平降低(P<0.05)。结论红参可以有效改善 DOR模型大鼠的内分泌,可能通过调控 PI3K/Akt/mTOR信号通路参与改善卵巢功能。     

MicroRNA-141通过PI3K/Akt/mTOR信号通路促进PCOS卵巢颗粒细胞增殖的机制研究

目的　研究microRNA-141（miR-141）对卵巢颗粒细胞增殖的影响,并探讨其在多囊卵巢综合征（PCOS）发生发展中的作用机制。方法　采用实时荧光定量PCR检测20例PCOS患者的卵巢组织、20例对照组的正常卵巢组织、人卵巢颗粒细胞KGN及人正常卵巢上皮细胞IOSE80中miR-141 mRNA表达水平;KGN细胞分为miR-141 minics组、LY294002+miR-141 minics组、雷帕霉素（rapamycin）+miR-141 minics组、NC组、对照组。采用MTT法和平板克隆实验检测各组细胞增殖和克隆形成能力,采用Western blot法检测各组细胞磷脂酰肌醇3-激酶（PI3K）/蛋白激酶（Akt）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路相关蛋白表达水平。结果　PCOS卵巢组织miR-141 mRNA表达水平显著高于正常卵巢组织,人卵巢颗粒细胞KGN miR-141 mRNA表达水平高于人正常卵巢上皮细胞IOSE80（P＜0.05）;miR-141 minics组、LY294002+miR-141 minics组及rapamycin+miR-141 minics组miR-141 mRNA表达水平均高于NC组和对照组（P＜0.05）;LY294002+miR-141 minics组及rapamycin+miR-141 minics组转染后24、48、72、96 h OD值和克隆形成率均低于miR-141 minics组,但高于NC组和对照组（P＜0.05）;LY294002+miR-141 minics组转染后p-Akt、p-mTOR蛋白表达水平低于miR-141 minics组,但高于NC组、对照组（P＜0.05）。rapamycin+miR-141 minics组转染后p-mTOR蛋白表达水平低于miR-141 minics组,高于NC组、对照组（P＜0.05）。结论　miR-141可通过激活PI3K/Akt/mTOR信号通路促进卵巢颗粒细胞的增殖。