The role of the PI3K-Akt signal transduction pathway in Autographa californica multiple nucleopolyhedrovirus infection of Spodoptera frugiperda cells

Many viruses activate the phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway, thereby modulating diverse downstream signaling pathways associated with antiapoptosis, proliferation, cell cycling, protein synthesis and glucose metabolism, in order to augment their replication. To date, the role of the PI3K-Akt pathway in Baculovirus replication has not been defined. In the present study, we demonstrate that infection of Sf9 cells with Autographa californica multiple nucleopolyhedrovirus (AcMNPV) elevated cellular Akt phosphorylation at 1 h post-infection. The maximum Akt phosphorylation occurred at 6 h post-infection and remained unchanged until 18 h post-infection. The PI3K-speci?c inhibitor, LY294002, suppressed Akt phosphorylation in a dose-dependent manner, suggesting that AcMNPV-induced Akt phosphorylation is PI3K-dependent. The inhibition of PI3K-Akt activation by LY294002 significantly reduced the viral yield, including a reduction in budded viruses and occlusion bodies. The virus production was reduced only when the inhibitor was added within 24 h of infection, implying that activation of PI3K occurred early in infection. Correspondingly, both viral DNA replication and late (VP39) and very late (POLH) viral protein expression were impaired by LY294002 treatment; LY294002 had no effect on immediate-early (IE1) and early-late (GP64) protein expression. These results demonstrate that the PI3K-Akt pathway is required for efficient Baculovirus replication.     

Akt/GSK-3β介导高糖上调肾小管上皮细胞Snail1的表达

 目的：观察高糖对原代肾小管上皮细胞Snail1和蛋白激酶B/糖原合成酶激酶3β（Akt/GSK-3β）信号通路的影响,探讨糖尿病肾病时Snail1表达的调节机制。方法：原代培养大鼠肾小管上皮细胞（RTECs）,随机分为正常糖对照组、高渗组和高糖组。Western blotting检测不同处理组 RTECs不同培养时点（30 min、2 h、12 h、24 h、48 h和72 h）Snail1、Akt1、GSK-3β、磷酸化Akt（p-Akt,Ser473）和磷酸化GSK-3β（p-GSK-3β,Ser9）蛋白的水平。RT-PCR检测Snail1、Akt1和GSK3β mRNA的表达。RTECs以磷脂酰肌醇3-激酶（PI3K）抑制剂LY294002（25 μmol/L）预处理50 min,再与高糖共同培养24 h,Western blotting检测上述指标蛋白的表达。结果：与正常糖对照组比较,高糖组RTECs Snail1和Akt1蛋白和mRNA的表达上调,p-Akt及p-GSK-3β蛋白表达增加,但总GSK-3β蛋白和mRNA表达无变化。以LY294002处理后,高糖组RTECs Snail1、p-Akt及p-GSK-3β蛋白表达水平较未处理高糖组明显下降,但LY294002不影响总Akt1和GSK-3β蛋白表达。结论：Akt/GSK-3β可能介导了高糖诱导的RTECs锌指转录因子Snail1的表达上调。     

PI3K/CTGF信号通路在TGF-β1诱导A549细胞表达collagen I过程中的作用

 目的： 研究磷脂酰肌醇3-激酶/结缔组织生长因子（PI3K/CTGF）信号通路在转化生长因子β1（TGF-β1）诱导人肺腺癌A549细胞表达I型胶原蛋白（collagen I）过程中的分子机制。方法： 体外培养A549细胞,予TGF-β1刺激,观察CTGF和collagen I的mRNA和蛋白表达及PI3K信号通路的活化;PI3K抑制剂LY294002预先处理A549细胞后,观察TGF-β1刺激下CTGF和collagen I mRNA和蛋白表达的变化;CTGF特异性siRNA干扰A549细胞中CTGF的表达后,观察TGF-β1刺激下collagen I mRNA和蛋白表达的变化和PI3K信号通路的活化。结果： TGF-β1可以诱导A549细胞中CTGF和collagen I的mRNA和蛋白表达以及PI3K信号通路的活化;PI3K特异性抑制剂LY294002可以部分逆转TGF-β1诱导的A549细胞中CTGF和Collagen I mRNA和蛋白表达的升高。干扰CTGF可以降低TGF-β1诱导的A549细胞collagen I mRNA和蛋白表达,而不影响PI3K信号通路的活化。结论： CTGF是TGF-β1/PI3K信号通路调控的即刻早期反应效应蛋白,参与了TGF-β1诱导的A549细胞中collagen I表达。     

Apolipoprotein E reduces food intake via PI3K/Akt signaling pathway in the hypothalamus

Apolipoprotein E (apoE) is a satiation factor. While central apoE administration reduces food intake, the specific intracellular signaling mechanisms activated by apoE remain largely unknown. Using primary cultured hypothalamic neurons, we demonstrated that apoE treatment (50 nM) elicited rapid activation of the phosphatidylinositol-3-kinase (PI3K)/Akt signaling cascade. Specifically, apoE induced the phosphorylation of Akt, peaking at 30 min, and the increased phosphorylation of Akt was significantly attenuated after pretreatment with LY294002 (50 μM), an inhibitor of the PI3K signaling pathway. To determine whether the activation of PI3K by apoE is required for the ability of apoE to reduce food intake, LY294002 (1 nmol) was infused into the 3rd-cerebral ventricle before injection of an anorectic dose of apoE. Consistent with our previous report, apoE (4 μg) exerted significant reduction of food intake in the 4-h fasted rats, compared with saline. Pretreatment with LY294002 significantly attenuated the potency of exogenous apoE to induce satiation, while the same dose of PI3K inhibitor by itself caused only a slight non-significant decrease of food intake. These results indicate that the activation of the PI3K/Akt pathway is necessary for the acute effects of apoE on food intake.     

ERK1/2和PI3-K通路在蛛网膜下腔出血大鼠海马区对神经细胞自噬的调控作用

目的:探讨细胞外信号调节激酶(extracellular regulated protein kinases,ERK1/2)和磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3-K)通路对大鼠蛛网膜下腔出血(subarachnoid hemorrhage,SAH)后神经细胞自噬的调控作用。方法:雄性SD大鼠160只,分为假手术(Sham)组、模型(SAH)组、ERK1/2抑制剂U0126组和PI3-K抑制剂LY294002组。采用二次注血法制作SAH大鼠模型,HE染色观察海马区神经细胞的形态变化;免疫组织化学染色法检测海马区磷酸化ERK1/2、PI3-K、自噬相关蛋白Beclin-1和微管相关蛋白1轻链(LC3)-II的表达实时荧光定量PCR检测海马区ERK1/2、PI3-K、自噬相关蛋白Beclin-1和微管相关蛋白LC3的表达。结果:SAH组海马区神经细胞存活率低于Sham组,ERK1/2、PI3-K、Beclin-1和LC3的表达水平高于Sham组(P0.05);U0126组和LY294002组海马区神经细胞存活率均高于SAH组,ERK1/2、PI3-K、Beclin-1和LC3表达均低于SAH组(P0.05);U0126组和LY294002组两组间海马区神经细胞存活率差异无统计学意义(P0.05),U0126组ERK1/2、Beclin-1和LC3表达水平低于LY294002组(P0.05),PI3-K表达水平高于LY294002组(P0.05)。结论:ERK1/2和PI3-K通路激活共同参与SAH后神经细胞自噬的调节,且以PI3-K通路更为主要。     

PI3K / Akt 通路调节LPS 诱导的小胶质细胞内HSPA12B 表达

目的:探讨PI3K/ Akt 通路对内毒素脂多糖(Lipopolysaccharide,LPS)诱导的小胶质细胞内热休克蛋白A12B(Heat shock proteins A 12B,HSPA12B)表达的影响。方法:体外培养小胶质细胞,并分三组处理:对照组、0.1 μg/ ml LPS 刺激组、PI3K/ Akt 通路抑制LPS 刺激组。Western blot 法检测小胶质细胞内HSPA12B 和Akt 磷酸化的蛋白水平表达,间接免疫荧光标记法检测HSPA12B 在小胶质细胞中的细胞表达定位。结果:LPS 刺激2 h 后,小胶质细胞内HSPA12B 和Akt 磷酸化的表达增加;应用LY294002 预处理后,LPS 诱导HSPA12B 蛋白水平表达显著抑制。免疫细胞荧光染色证明小胶质细胞LPS 组HSPA12B 核周荧光强度明显增强,LY294002 预处理组HSPA12B 荧光强度明显减弱。结论:PI3K/ Akt 途径参与调控LPS 诱导小胶质细胞HSPA12B 表达。     

PI3K/Akt/eNOS信号通路在葛根素抑制ox-LDL诱导的血管内皮细胞组织因子表达中的作用

目的:探讨磷脂酰肌醇3-激酶/蛋白激酶B/内皮型一氧化氮合酶(PI3K/Akt/eNOS)信号通路在葛根素(puerarin)抑制氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导的血管内皮细胞组织因子(tissue factor,TF)表达中的作用。方法:实时荧光定量PCR检测TF的mRNA表达,Western blot检测TF和Akt的蛋白表达,硝酸盐还原酶法检测一氧化氮(nitric oxide,NO)含量。结果:与对照组相比,ox-LDL孵育内皮细胞后,内皮细胞TF的mRNA和蛋白表达升高,Akt蛋白磷酸化水平降低,细胞内NO产生减少;而葛根素预孵育内皮细胞1 h后,再用ox-LDL孵育,内皮细胞TF mRNA和蛋白表达下降,Akt蛋白磷酸化升高,细胞内NO产生增多;PI3K抑制剂LY294002和葛根素共同预孵育内皮细胞1 h后,再用ox-LDL孵育,内皮细胞TF的mRNA和蛋白表达升高,Akt蛋白磷酸化降低,细胞内NO产生减少;eNOS抑制剂NG-硝基-L-精氨酸甲酯(L-NAME)和葛根素共同预孵育内皮细胞,也明显阻断葛根素对ox-LDL诱导的内皮细胞TF mRNA和蛋白表达、细胞Akt蛋白磷酸化和细胞内NO产生的作用。结论:葛根素可通过上调PI3K/Akt/eNOS信号通路抑制ox-LDL诱导的人脐静脉内皮细胞TF mRNA和蛋白表达。     

High-glucose and advanced glycosylation end products increased podocyte permeability via PI3-K/Akt signaling

Regardless of the underlying disease, the proteinuric condition demonstrates ultrastructural changes in podocytes with retraction and effacement of the highly specialized interdigitating foot processes. To investigate how high-glucose (HG) and advanced glycosylation end products (AGE) induce podocyte phenotypical changes, including quantitative and distributional changes of zonula occludens (ZO)-1 protein and search for the signaling mechanisms, we cultured rat glomerular epithelial cells (GEpC) and mouse podocytes under: (1) normal glucose (5 mM, control); (2) HG (30 mM); (3) AGE-added; or (4) HG plus AGE-added conditions. HG plus AGE increased the permeability of monolayered GEpCs and induced ultrastructural separation between confluent GEpCs. ZO-1 moved to inner actin filament complexes in both AGE- and/or HG by confocal imaging. HG plus AGE-added condition also decreased ZO-1 protein amount and mRNA expression compared to normal glucose or osmotic control conditions. We could also confirm the induction of RAGE (receptor for AGE) and PI3-K/Akt signaling pathway by AGE and HG. In addition, LY294002, a PI3-K inhibitor, could prevent the quantitative and distributional changes of ZO-1 and RAGE and the increased permeability induced by HG and AGE. These findings suggest that diabetic conditions induce the podocyte ZO-1 changes via RAGE and PI3-K/Akt signaling, leading to increased permeability.     

Down-regulation of the PI3-kinase/Akt pathway by ERK MAP kinase in growth factor signaling

The ERK MAP kinase and PI3-kinase/Akt pathways are major intracellular signaling modules, which are known to regulate diverse cellular processes including cell proliferation, survival and malignant transformation. However, it has not been fully understood how these two pathways interact with each other. Here, we demonstrate that inhibition of the ERK pathway by the MEK inhibitor U0126 or PD98059 significantly potentiates EGF- and FGF-induced Akt phosphorylation at both Thr308 and Ser473. We also show that hyperactivation of the ERK pathway greatly attenuates EGF- and FGF-induced Akt phosphorylation. Furthermore, the enhanced Akt phosphorylation induced by U0126 is inhibited by the PI3-kinase inhibitor LY294002, and is accompanied by the up-regulation of Ras activity. These results suggest that the ERK pathway inhibition enhances Akt phosphorylation through the Ras/PI3-kinase pathway. Thus, our results demonstrate that the ERK pathway negatively modulates the PI3-kinase/Akt pathway in response to growth factor stimulation.     

Insulin-like growth factor-1 induces the phosphorylation of PRAS40 via the PI3K/Akt signaling pathway in PC12 cells

Insulin-like growth factor-1 (IGF-1) is a polypeptide tropic factor that plays an important role in the survival and differentiation of both neuronal and non-neuronal cells. Numerous studies have demonstrated that IGF-1 promotes neuronal cell survival via the PI3K/Akt signaling pathway. Proline-rich Akt substrate of 40kDa (PRAS40) is a recently discovered downstream target of Akt. However, the relationship between IGF-1 and PRAS40 is not known. In this study, we characterized the phosphorylation of PRAS40 induced by IGF-1 in PC12 cells and explored the signaling pathway responsible for the effect of IGF-1. IGF-1 induced the phosphorylation of Akt at Thr473 and PRAS40 at Thr246 in PC12 cells. The phosphorylation of Akt and PRAS40 induced by IGF-1 (100ng/ml) was inhibited by the phosphatidylinositide 3-kinase (PI3K) specific inhibitor LY294002 (50μM), while no inhibitory effect was observed for a MAPK kinase pathway specific inhibitor PD98059 nor a p38 MAPK inhibitor PD169316, suggesting that the phosphorylation of PRAS40 induced by IGF-1 is mediated by the PI3K pathway in PC12 cells and primary cultured neurons. In further support this hypothesis, an Akt kinase specific inhibitor, Akt inhibitor VIII, attenuated IGF-1-induced phosphorylation of PRAS40 at the concentration that blocked the phosphorylation of Akt induced by IGF-1. Taken together, these data demonstrate that IGF-1 stimulates the phosphorylation of PRAS40 at Thr246 in neuronal cells and the effect of IGF-1 is mediated, at least in part, by the PI3K/Akt signaling pathway.     

Serotonin-induced growth of pulmonary artery smooth muscle requires activation of phosphatidylinositol 3-kinase/serine-threonine protein kinase B/mammalian target of rapamycin/p70 ribosomal S6 kinase 1

We have previously found that both mitogen-activated protein kinase (MAPK)- and Rho kinase (ROCK)-related signaling pathways are necessary for the induction of pulmonary artery smooth muscle cell (SMC) proliferation by serotonin (5-hydroxytryptamine [5-HT]). In the present study, we investigated the possible additional participation of a phosphatidylinositol 3-kinase (PI3K)/serine-threonine protein kinase B (Akt)/mammalian target of rapamycin (mTOR)/p70 ribosomal S6 kinase (S6K1) pathway in this growth response. We found transient activation of Akt (Ser473) and more prolonged activation of S6K1 by 5-HT. Inhibition of PI3K with Wortmannin and LY294002 completely blocked these activations, but not that of MAPK or the ROCK substrate myosin phosphatase targeting subunit. Similarly, inhibition of MAPK and ROCK failed to block the Akt activation. Inhibition of Akt with NL-71-101 and downregulation of Akt expression with Akt small interfering RNA blocked 5-HT-induced S6K1 phosphorylation. Wortmannin, LY294002, and NL-71-101 dose-dependently inhibited 5-HT-induced SMC proliferation. 5-HT stimulated mTOR phosphorylation and the mTOR inhibitor, rapamycin, blocked activations of S6K1 and S6 ribosomal protein, and inhibited 5-HT-induced SMC proliferation. Akt phosphorylation and cell proliferation were also blocked by the antioxidants, N-acetyl-l-cysteine, Ginko biloba 501, and tiron, the reduced nicotinamide adenine dinucleotide phosphate oxidase inhibitor, diphenyleneiodonium, and the 5-HT2 receptor antagonists ketanserin and mianserin, but not by the 5-HT serotonin transporter or 5-HT 1B/1D receptor antagonists. We conclude from these studies that a parallel PI3K- and reactive oxygen species-dependent Akt/mTOR/S6K1 pathway participates independently from MAPK and Rho/ROCK in the mitogenic effect of 5-HT on pulmonary artery SMCs. From these and other studies, we postulate that independent signaling pathways leading to 5-HT-induced SMC proliferation are initiated through multiple 5-HT receptors and serotonin transporter at the cell surface.     

抑制Akt通路对前列腺癌细胞株糖酵解影响的探讨

目的:探讨LY294002和氯化钴对PC3和DU145细胞糖酵解的影响及可能的机制.方法:不同浓度的LY294002、氯化钴和LY294002联合氯化钴处理PC3和DU145细胞,检测细胞的葡萄糖消耗、Akt的磷酸化、HIF-1α的蛋白表达和几个基因的mRNA表达.结果:LY294002减少PC3细胞Akt的磷酸化、H...     

盐酸小檗碱通过PI3K/Akt信号通路促进小鼠前成骨细胞MC3T3-E1的分化

目的:研究盐酸小檗碱对小鼠前成骨细胞系MC3T3-E1分化与矿化的调控作用及其机制。方法:MC3T3-E1细胞给予不同浓度(0、1、5、10和20 mg/L)的盐酸小檗碱刺激3 d,CCK-8法检测细胞活性。不同浓度的盐酸小檗碱分别干预3 d和7 d,检测细胞碱性磷酸酶(ALP)活性。进一步将实验随机分为4组:对照组、盐酸小檗碱组、盐酸小檗碱+LY249002(PI3K/Akt通路抑制剂)组及LY249002组。干预2 d后,采用real-time PCR检测成骨细胞分化相关因子ALP、骨钙素(OCN)、骨桥蛋白(OPN)及Runt相关转录因子2(Runx2)的mRNA表达情况,采用Western blot检测PI3K/Akt信号通路相关蛋白p-Akt的表达水平。将MC3TC-E1细胞用矿化培养基诱导21 d,茜素红染色检测其矿化情况。结果:与对照组相比,不同浓度的盐酸小檗碱对细胞活性的影响没有明显差异;不同浓度的盐酸小檗碱处理MC3T3-E1细胞后ALP活性有不同程度升高。Real-time PCR结果表明,盐酸小檗碱(5 mg/L)促进ALP、OCN、OPN及Runx2的mRNA表达(P 0. 01),而LY294002能抑制这些分化相关因子的表达。Western blot检测结果表明,盐酸小檗碱(5 mg/L)促进p-Akt蛋白的表达(P 0. 01),其作用被LY249002抑制。茜素红染色发现盐酸小檗碱组矿化明显,但LY294002能抑制盐酸小檗碱的促进作用。结论:盐酸小檗碱可以促进小鼠前成骨细胞的分化与矿化,其机制可能与其激活PI3K/Akt信号通路有关。     

促红细胞生成素通过PI3-K/Akt信号通路抑制血管紧张素II诱导的新生大鼠心脏成纤维细胞增殖

目的： 探讨促红细胞生成素（EPO）对血管紧张素Ⅱ(Ang Ⅱ)诱导的心脏成纤维细胞（CFs）增殖的影响,以及磷脂酰肌醇-3-激酶/蛋白激酶B（PI3-K/Akt）信号途径和一氧化氮合酶（NOS）的作用。方法: 应用胰酶和胶原酶双酶和差速贴壁法分离培养新生大鼠CFs细胞,应用EPO、Ang Ⅱ、PI3-K抑制剂LY294002、NOS抑制剂L-NAME不同因素干预。细胞计数和MTT法作出CFs的生长曲线,检测CFs的增殖。化学酶法检测CFs培养液中的一氧化氮（NO）浓度以及总NOS和其亚型的活性。Western blotting检测Akt、p-Akt、内皮型一氧化氮合酶（eNOS）和诱生型一氧化氮合酶（iNOS）蛋白的表达。结果: Ang Ⅱ促CFs增殖的作用显著。EPO剂量依赖性的增加CFs培养液中的NO浓度,同时剂量依赖性的抑制Ang Ⅱ诱导的CFs增殖。在给药后第4 d,和单纯的Ang Ⅱ相比,浓度为5×103 U/L、1×104 U/L和2×104 U/L EPO对CFs增殖的抑制率分别达到了24.4％、41.5%和50.5%。EPO显著提高Akt的磷酸化水平,促进eNOS蛋白的表达。应用PI3-K抑制剂LY294002和NOS抑制剂L-NAME均能使培养液中的NO浓度也随之下降,EPO抑制Ang Ⅱ诱导CFs增殖的作用均被阻断。但LY294002同时阻断了eNOS蛋白表达,而L-NAME对eNOS没有影响。结论: EPO可剂量依赖性的抑制Ang Ⅱ诱导的新生大鼠CFs的增殖。其作用机制是通过激活PI3-K/Akt信号途径促使CFs中eNOS表达来促进NO的生成,从而抑制CFs的增殖。     

骨髓间充质干细胞分泌基质细胞衍生因子1保护心肌细胞

背景：有研究显示表达CXCR4的干细胞能够沿着基质细胞衍生因子1的浓度梯度迁移到心肌梗死部位再生心肌和血管而改善心脏的功能。 目的：探索间充质干细胞通过其分泌的基质细胞衍生因子1对心肌细胞的保护作用。 方法：收集培养2 d的间充质干细胞条件培养基。在缺氧条件,利用基质细胞衍生因子1受体CXCR4阻断剂AMD3100或PI3-K/Akt途径阻断剂LY294002预处理H9C2细胞后,利用AnnexinV/PI双标法流式细胞术分析间充质干细胞条件培养基作用下H9C2细胞凋亡的变化;Western blotting分析H9C2细胞磷酸化Akt蛋白的表达;RT-PCR分析间充质干细胞基质细胞衍生因子1的表达。 结果与结论：RT-PCR结果显示间充质干细胞表达基质细胞衍生因子1,Western blotting结果显示间充质干细胞条件培养基增加了H9C2细胞磷酸化Akt蛋白的水平。AnnexinV/PI分析发现间充质干细胞条件培养基明显降低了H9C2细胞缺氧复氧后的凋亡,且这种抗凋亡作用能被CXCR4阻断剂AMD3100或PI3-K/Akt途径阻断剂LY294002所阻断。说明间充质干细胞通过其分泌的基质细胞衍生因子1通过激活PI3-K/Akt途径保护H9C2细胞,增加H9C2细胞的幸存能力。     

Nephrin稳定血管紧张素Ⅱ诱导的足细胞细胞骨架改变的机制研究

 目的：研究足细胞裂孔膜分子nephrin调节血管紧张素Ⅱ（AngⅡ）诱导的足细胞骨架分布改变的分子机制。方法：用AngⅡ及AngⅡ受体拮抗剂氯沙坦或Akt抑制剂LY294002刺激足细胞,FITC-phalloidin染色标记F-actin,分析足细胞骨架运动。Real-time RT-PCR、RT-PCR和Western blotting检测nephrin mRNA和蛋白表达。转染nephrin全长表达质粒（pcDNA3.1-mNPHS1）,建立稳定转染足细胞系,Western blotting检测转染细胞的Akt磷酸化水平,FITC-phalloidin染色分析高表达nephrin对F-actin分布影响。结果：AngⅡ和LY294002刺激后,足细胞的F-actin重组,应力纤维减少,形成F-actin外周环。氯沙坦显著抑制F-actin重排。AngⅡ刺激后nephrin mRNA和蛋白表达显著降低,Akt磷酸化水平降低。pcDNA3.1-mNPHS1转染显著上调足细胞Akt磷酸化水平,促进足细胞短线状足突形成,部分抑制AngⅡ诱导的骨架重排。结论：PI3K/Akt是nephrin和AngⅡ的共同下游通路。Nephrin能通过PI3K/Akt途径部分稳定AngⅡ诱导的足细胞细胞骨架改变。     

Adipophilin通过PI3K/Akt通路促进细胞内脂质蓄积

目的:探讨adipophilin促进细胞内脂质蓄积的可能机制,为动脉粥样硬化的防治提供参考。方法:分别通过q PCR、Western blot和油红O染色观察氧化型低密度脂蛋白(ox LDL)处理RAW264.7细胞不同时间后,细胞内Akt、p-Akt和adipophilin的蛋白水平及脂质蓄积情况;并检测PI3K/Akt抑制剂LY294002处理后,上述指标的变化;HEK293细胞过表达adipophilin后,检测Akt的活性;并用免疫共沉淀实验检测adipophilin与Akt之间的相互作用。结果:ox LDL处理细胞后,随着时间的延长,脂滴增多,Akt被活化,adipophilin表达增加,而LY294002处理则可抑制上述变化;过表达adipophilin后,p-Akt水平增高,但adipophilin与Akt之间未见直接相互作用。结论:Adipophilin可通过PI3K/Akt途径促进细胞内脂质蓄积,但可能不是通过直接相互关系发挥作用的。